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Skeletal Muscle - Science topic
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Questions related to Skeletal Muscle
Hello,
I have no prior experience with growing cell line nor differentiation toward specific cell type. Currently I have C2C12 cell line which I need to grow and then differentiate it towards skeletal muscle cells.
Can you please provide me with as much as possible information, SOP, tips and tricks?
Thank you
Hi everyone, I am looking for a human skeletal muscle cell line which however requires differentiation times of 7-10 days and which can grow in horse serum. Could someone recommend me an easily manageable line?
Thank you a lot
I am working on skeletal muscle synthesis, and I did Western to see p70s6k.
I expected I could see the p70s6k band around 70 kDa, but I saw two unexpected bands below (attached).
I am not sure how to interpret the bands.
Western information that I did is the following:
10% gel__Running 50v for 30min and then 100v for 2h__Transfer membrane (PVDF) for overnight in cold room (36v)__Cell signaling Primary (rabbit)
I would appreciate it if you leave any comment on that.
Sincerely,
Sonny.
We have infected mice with an AAV-EGFP and we were hoping to directly visualize the fluorescent signal in frozen muscle sections without antibody staining.
Any suggestions would be greatly appreciated.
Dear sir,
I am going through your paper titled " Micromechanical modelling of skeletal muscles based on the finite element method " . You have mentioned force velocity characteristics with equation (numer 9 and plotted the graph (figure 4)).In that relation kc , ke are the curvature and d is the offset of the eccentric function.
My question is kc,ke , d same for all kind of muscles ? if not how to get the values of kc,ke , d values for different muscles?
Could you please reply me sir.
We are trying to isolate nuclei from mouse skeletal muscle but at the end of the process, the clumping of nuclei was still present in the slide. We were not getting good-quality of nuclei for library preparation and further genomics study.
As a resuspended buffer, I used RNAse inhibitor, 2% BSA and 1X PBS buffer. So, I would like to know how to minimize/remove these clumping nuclei and get better nuclei quality at the final stage after the resuspension process. Please anyone can help me how to solve this problem?
Thanks in advance
Skeletal muscles are stained with the blue color but other half of the picture was stained with red color. Can someone tell me what kind of tissue or cell is it?

I'm looking for a protocol for measure SDH activity in skeletal muscle homogenate. Are there any protocols that do not include NaCN or KCN?
working on analyzing skeletal muscle cells exit form myogenic differentiation.
During examination of transmission electron microscopy images of skeletal muscle biopsies from patients suffering from different myopathies, sometimes I find somewhat unusual mitochondria with white spots. Initially, I assumed that is some sort of oxidative stress damage or even artifact, yet now I think I could be wrong; usually, in patients with frequent "white spotted" mitochondria, I find more pronounced pathological changes in mitochondria (such as paracrystalline inclusion or onion-like cristae) in other fibers, sooner or later.
That made me think, so I've tried to find similar images in other studies, but to no avail. I have no reasonable explanation as for now. Any ideas?

Hi fellow researchers, has anyone ever worked with AICAR to induce activation of AMPK in L6 skeletal muscle cells? So far I tried to activate it by adding AICAR (either 0.1mM or 1mM) in DMEM with 1% pen strep to L6 myotubes. This was done after serum starvation for about 24hours. However, the AMPK was not activated when I compared with the untreated group. Has anyone ever activated it in these types of cells?
I would like to understand how different load on the muscle affects the development of bone tissue, the presence of tubercles on the bones, thickening, etc.
I'm currently working with Western Blotting technique for human skeletal muscle. However, when i use the Cell Signaling antibody (#8164 and #14179) no image were reveled. Apparently, i dont have problems with WB protocol, once others antibody brands (e.g., Santa Cruz and ABCAM) works for others proteins. In addition, the lysate protocol and the WB protocol used were performed according Cell Signaling recomendations.
PFK (D4B2 - 8164S ) - Rabbit mAb - Lot: 5
HIF-1alpha (D2U3T - 14179S) - Rabbit mAb - Lot 4
Has anyone had similar problems with Cell Signaling antibody? If yes, how to solve this problem?
Dear colleagues,
I was wondering whether there are any ways or any softwares that I can use to analysis the muscle cross sectional area in my H&E histology images?
I tried to use ImageJ thresholding, unfortunately it does not work efficiently for me. Thus, I was wondering whether there are any currently established methods.
Thank you very much in advance.
I am working on natural and synthetic insulin mimetics and insulin potentiating agents. I am assessing the effect compounds on glucose uptake in L6 myotubes, for that I want use Insulin as positive control. However I am doubtful that human insulin can work on cell lines derived from rat.
Any clarifications is appreciated. Thanks in advance
For the past few weeks we've been using three different cell types on the Fura 2 calcium experiment including iPSC's, RenCells, and human skeletal muscle cells. We loaded our cells with 2.5 uM of Fura 2 in HBSS + 1% BSA. We incubated the cells in both 37 degrees or at RT for about 30 minutes and hydrolyzed for about 20 minutes in HBSS. Using the Olympus microscope and MetaFluor software we were able to see the cells with the fluorescent microscope indicating the loading was successful, but saw no response when exposed to the drugs we used. As a positive control test we used 5mM glutamate or 20uM ionomycin and also saw no response. I appreciate if anyone could give any insight on potential issues or solutions we can take to successfully run this experiment.
I used proteinase K as a protein digestion step before using a Qiagen AllPrep DNA/RNA extraction kit. I have used it before on WT murine heart and skeletal muscle tissues and got good yield. Today, when I used it on my experimental samples, I got virtually no DNA or RNA.
I added the appropriate amount of ProK to my samples and incubated for 10 mins at 55 degrees as recommended. Then, I spun them down and left the samples on ice while I ate lunch (~45 minutes). This break was the only difference in my protocol today versus my first test run.
I was hoping that icing my samples would inactivate ProK, but I believe that it remained active and digested all of my DNA and RNA.
what dye I can use to look for Ca2+ distribution in muscle cell?
Hello, i did H&E staining using cryosection.
However, there are so many holes in myofiber.
Here is my protocol.
No fixation for myofiber staining
1. Dissection - skeletal muscles
2. Embed into O.C.T using liquid nitrogen
3. After completely frozen, stored at -80c before using
4. Put it into -20c cryostats 20 mins before sectioning.
5. Cut to 10μm thickness
However, it is really hard to cut without getting holes, even though the blade is brand new.
Is the problem of water crystalization or freezing time?
Please help me.
Hello, everyone. I would like to determine the fiber type of each skeletal muscle fiber based on its SDH and ATPase activity.I used the dry ice-hexane method to freeze freshly isolated skeletal muscle because the liquid nitrogen freezing method produced bubble artifacts. Although these methods were used to evaluate by enzyme activity, there is unavoidable bubble artifact.
If it is difficult, I will prepare frozen sections after saturation with normal sucrose and use an antibody-based staining method for each muscle fiber.
If you have any knowledge, I would appreciate it very much.
Thank you in advance.
I was wondering if there is a reason to use one method over the other. Some literature used acetone and ethanol (3:1), while others used acetone and methanol (1:1). Which mixture do you think leads to better results? And why?
I'm staining for skeletal muscle proteins (e.g. MHCI, COXIV, PLIN proteins ...).
i don't get any DNA a the end of the protocol, i tried BEB buffer modification too but nothing works, any clue?
I am looking to submit a brief review for publication. Brief reviews are similar to mini reviews in their requirements. However, I am looking to see which journals best fit the scope of my paper. It's related to aging, cardiovascular health and skeletal muscle performance. The only journal that I have found so far that says they accept brief reviews that also has submission guidelines for a brief review is: International Journal of Sports Physiology and Performance. I would like to see what other options I might have beyond this journal. Any help would be appreciated. Thank you.
I am woking with type II diabetes and doing assay on L6 skeletal muscle cell line to understand the glucose uptake by using on-cell western method. But I am confused Do i need to protein quantification for On-cell western?
Could someone help explain to me why have researchers mostly focus on the GSP muscles ( gastrocnemius, soleus and plantaris) , but not the tibialis anterior (TA) in the hindlimb suspension model?
Hi everyone,
I am working on a grant on muscle Na+,K+-ATPase activity in human subjects. However, our lab does not have a protocol on the assessment of Na+,K+-ATPase activity in skeletal muscle homogenates. Although I could find some info in relevant papers, it is still hard for me to have a precise idea about the procedure and the reagents/devices needed. Could you let me know where I can find a step-by-step protocol?
Thank you in advance!
Shelly
Ø Before starting the muscle biopsy procedure, pour 100 mL (approximately 1/3 of beaker) isopentane (2-methybutane) to a glass beaker or a steel beaker.
Ø Suspend the beaker in a bath of liquid nitrogen (either in a larger beaker or Styrofoam box) and leave until the isopentane ice crystals start to form on the bottom of the beaker and on the sides of the beaker. When this occurs, isopentane has reached optimal freezing temperature (-150 °C). Time taken averages between 3-5 minutes.
Ø Following the skeletal muscle biopsy, remove the excess moisture by dabbing the tissue on the filter paper. Place muscle in proper orientation. Cover it in OCT (Optimum Cutting Temperature compound), and lay it flat on a labeled disposable freezing mold. Freeze the muscle for about 10-12 seconds (Being very small muscle tissue)
Ø Transfer the frozen tissue on to dry ice and let the isopentane evaporate( approx. 20minutes) and wrap the tissues in aluminum foil and store them temperature (-70 °C to -80 °C) .
I recently carried out ELISA on L6 skeletal muscle cell lysates using Elabscience GLUT 4 Elisa kit in order to determine the GLUT 4 expression in the cells when treated with different treatments. However, I have been getting no expression in all my treatments including the positive controls although, my standards for the Elisa produced a good curve. I tried to use different methods to extract the cell lysates and i confirmed that the treatments were done properly but there isn't any difference when I repeated the ELISA. All the results from the ELISA appear negative. May I know if anyone has used ELISA before to determine GLUT 4 expression in these cells? Which extraction method was used? or perhaps is there another better method to determine the expression?
I would like to identify the subcellular localization of nitrate storage in skeletal muscle.
perhaps the same mechanism governs lateral force transmission in a single skeletal muscle cell? (Sybil Street in J Cell Physiol and Science)
I used this kit for measuring Total TG content in rat Plantaris muscle. However, I have some questions about the calculation of the final TG concentration.
It is highly appreciated if anyone can help me.
I am looking for a non-NMR based (preferably) method of analyzing glycogen synthesis in the skeletal muscle and heart. Can anyone share a protocol where one has used radiolabeled glucose incorporation into muscle glycogen in vivo?
Usually skeletal muscle after injury undergo regeneration process following by all key phases of degeneration, inflammation and so on so until regeneration of fibers. For example we usually identify regenerating myofibers on sections by centrally located nuclei, however I am curious about how long (time/days) it take further for the regenerating myofibers to be mature, when the centrally located nuclei will already be moved to periphery of membrane as fibers 'll be mature.
Any valuable suggestions would be greatly appreciated, please.
Thank you..!
Spinal Muscular Atrophy is inherited disorder. Defective gene is the SMN1 gene which codes for SMN protein. Defect in this gene or its absence can cause death of motor neuron in the spinal cord leading to weakness of voluntary muscles.
We are trying to probe for the RpS6 protein which typically shows up around 32 kDa on a western blot. We have two questions regarding probing for this protein:
1. We recently ran the protocol for soleus muscle, and the protein showed up around 15 kDa. What could be the cause of this? Could it be that the protein broke down before we probed for it, or that this is the result of nonspecific binding? Any speculation is welcome.
2. When we probed for the protein in another muscle (gastrocnemius), no band showed up at all. RpS6 is a ribosomal protein, so we figured it would be present in this muscle and that there was an error in probing for it. We are using PVDF. Thanks.
I am trying to measure differences in eNOS activity in skeletal muscle in sedentary and exercised mice, following the conversion of 3H-arg to 3H-cit, and assaying the samples with and with out L-NAME to get eNOS activity. However, I am having multiple problems, from signal no different from the blank (even with ~300 ug protein, fresh 1.25 mM L-NAME, fresh buffer components (including FAD, FMN, BH4, NADPH, CaM) and plenty of CaCl2), to no difference in activity with and without L-NAME, to huge variance between replicates (my pipetting is not that bad!!). I have 10 uM Arg total (1 uM 3H-Arg) in the assay, and allowing the samples to react for 30 min at 37C. I stop the reaction with NaAcetate pH 5.5 / 5 mM EDTA, and then pass the solution over dowex AG 50W x8 (Na form) to bind unreacted Arg in the solution. I am happy to share my protocol with anyone who can help me figure out what I am doing wrong!! Thanks for any and all help!
I have purchased the Horse skeletal muscle Myoglobin (M0630) and Sodium dithionite (157953) from Sigma Aldrich. During my experiment, Mb (concentration 11 μM to 17 μM) gave its absorbance at 409nm in UV-Vis spectrophotometer but after adding the 100μl of 100 μM Sodium dithionite, the shift in the soret maximum from 409 nm to 435nm was not observed. Rather, the intensity of the Mb peak was reduced. An even higher concentration of sodium dithionite gave further reduction only in the soret band of Mb obtained in the 409 nm. The buffer I used to dissolve the components was 0.1M Phosphate buffer (pH-7.0) and incubation time was 5 to 30 min.
I have mostly been using the immortalised C2C12 murine cell line for in vitro skeletal muscle atrophy studies. In my lab, I used serum-free DMEM for 24 hours to stimulate muscle atrophy in C2C12 myotubes, which was validated via upregulation of Atrogin-1 and MurF1 through RT-qPCR. I'm planning on using the human immortalised myoblast cell line LHCN-M2 for validation. Reading the protocol I found serum-free media is required for differentiation. So my question is what methods can I use to mimic atrophy in LHCN-M2 myotubes?
I will be freeze-drying small skeletal muscle samples (~60mg) per sample using a Virtis SP Scientific Benchtop Pro Freeze Dryer. I've had difficulty findings a reference protocol to determine the optimal length and would love input. There is seemingly plenty amongst various animal models using larger samples (200-300mg) running them for ~24h, but I'd like to save time if possible.
Has anyone seen any such methodology and with the above device if possible?
My OD value had a huge variation between each trial, even blank one. The worst thing is blank two has higher OD than the corresponding sample no matter if it was diluted or not. The repeatability of the assay kit is not good. My protocol is as follows.
I am running an oxyblot on homogenized skeletal muscle tissue lysate. When I added all the reagents from the millipore oxyblot kit, the samples turned yellow and I got a grainy white precipitate in the mixture. This happened in samples with the DNPH mixture and the control mixture. Is this a normal reaction and if not, any recommendations to prevent this?
Dear all,
I am looking for reliable antibodies for the adrenergic receptors alpha1D and alpha2A, to use in Western Blotting on human skeletal muscle tissue. Anyone with experience with good antibodies from similar tissue preferably from humans?
I have come across many different antibodies, all of which has since been removed from the supplier, and I am therefore looking for some good experiences or references.
Hi all,
I´m stimulating skeletal muscle cultures prepared from human donors. After one week of differentiation I use a C-Pace system with the following conditions: 2-5 Volts, 2 ms, 1 Hz. I use 12 wells culture plates. Although the electrical conditions are not extreme compared to the available literature (for the same commercial system), my cells are damaged by the stimulation. Even only 2 hours of stimulation reduce the number of cells and induce vacuoles and vesicles (visible with transmitted light). I don´t know if the type of culture plate (previous reports use 6 or 24 wells plates) could explain the deleterious effect of the stimulation.
Any experience/input would be appreciated.
Thanks in advance
I am looking to quantify both human serum and skeletal muscle homogenate betaine concentrations via ELISA or colorimetric assay kits. Has anyone had a good experience with any products accomplishing this?
The assay we were planning to use recently became discontinued without further plans to restock. Any help is greatly appreciated!
I am planing to apply EMS on upper extremity muscles and observe muscle using sEMG and MMG. I am wondering how to do the normalization in case of EMS? is it same as in the case of normal sEMG and MMG i.e. dividing the sub-maximal signals with the signals obtained during MVC?
We are trying to prepare organoids from myoblasts. Could you help us about the typr of additional stromal cells or any useful methods to built them?
Thanks!
ross
creatine digestion, absorption and circulation. How?
Is C digested? Or taken in as a whole?
How is C absorbed by the enterocyt? Are there transporters?
How is C leaving the enterocyt? Sometimes structures get altered in there.
How is C travelling through the blood circulation?
Does C go to the liver?
How much C gets into the circulation comparred to ingestion?
Think about C and skeletal muscle there is enough information.
Thanks a lot!
Hi Researchers,
We use the trizol/chloroform phase separation extraction method and have been consistently ending up with RNA samples with high (3-5) 260/280 ratios. I have not been able to find much literature for troubleshooting this as most contamination issues/solutions are for low ratios and shifted peaks. It only seems to happen with skeletal muscle and I frequently end up with white, very sticky pellets that need to be manually removed from the side of the tube with a sterile tip during ethanol washes. Does anyone have any suggestions?
I will attach the protocols but in brief: we make our own RNAsol, homogenise tissue in 500uL using hand-held pestles and continue with the chloroform/isopropranol phase-separation extraction method. As skeletal muscle is a fibrous material, I added a 10min 4 degree spin at 12000g prior to chloroform addition to pellet ECM etc. but most recently, for EDL muscle, this did not make a difference. RNA samples generally generate good peaks (low left shoulder, peak at 260) but higher than ideal ratios. We are concerned this causes issues with qPCR. Our cleanup protocol always produces RNA samples with ratios in the recommended range but this makes extraction more time-consuming and lower yield. Using a kit has also addressed this issue but that is expensive and I worry about using this method for smaller tissues weights (ie. EDL vs TA) We do not understand what the contaminant is...
Thanks for your help!
I am performing macrophage staining in the oral mucosa using an IFA protocol. There are oval-shaped structures on my scans that have background stain/autoflorescence. After doing some research, I found they are either cross-sections of skeletal muscle or adipose tissue. They sometimes appear ordered and clumped like they are skeletal muscle cross-sections, while other times they are more sporadic as if they would be adipose tissue. I am having difficulty identifying which is which. Do both tissue types usually autofloresce? If so, how can I differentiate between them while looking at my scans? Thank you!
I need to perform live/dead stain on fresh skeletal muscle slide but I am struggling to cut the fresh skeletal muscle to desire thickness.
The plan is to embed the fresh skeletal muscle in 4% agarose. I tried to dry it as much as I could with Kimwipe; also tried to dip it in petri dish of agarose gel, move around then move it to the moulding base (aim is to remove as much water as possible). Unfortunately, the muscles still tend to fall apart during vibratome sectioning. I am not sure if it's due to the residual liquids from muscle or it's just difficult to cut due to the epimysium / perimysium.
I am just wondering if anyone has any experience / tips on cutting fresh skeletal muscle with vibratome, and what thickness you managed to achieve. Thank you.
Dear Researchers,
We have generated an inducible satellite cell-specific knockout model (iIpax7-cre). Our aim is to confirm a negative impact of our KO on muscle development. To do so, we are inducing KO at ~8wo and attempting to injure muscle and induce expansion of the target protein-depleted satellite cells using cardiotoxin injection.
Our lab is new to this and we are observing less effect on muscle development markers in our controls following cardiotoxin injection than we would expect. We are aware of significant batch variation issues in the literature/through collaborators. We have read about other muscle injury models.
Aside from this, I am unsure whether this is the best method to address our aim. Could any researchers with experience in the area please provide suggestions?
I knocked out a gene which is expressed in skeletal muscle, one is hsp40 and another one is a co-chaperone of hsp90a1. Previous study showed that hsp90a1 is specially expressed in skeletal muscle, which involved in sarcomere protein assembly.
I am using QIAamp Mini Kit for DNA extraction and having relatively low yields from heart, skeletal muscle and adipose tissue (around 2 ug). I tried to increase the amount of proteinase K and to extend the incubation time at 56 deg C to get complete tissue digestion but it didn't help much. Any suggestions?
A patient with hereditary desminopathy (Thr341Pro DES mutation in a heterozygous state) was recommended to refuse toothpaste. He continued to brush his teeth twice a day with a toothbrush with only water. As a result, within one month we noted a significant increase in strength and muscle mass in this patient. The patient did not take any medications during this period. After 30 days, the muscle condition returned to its original level. How can this positive effect be explained?
I am working on C2C12 skeletal muscle cell line in 6 well plate. I want to do Transmission electron microscopy (TEM) of my sample with cell line . how much sample need or where is possible in India.
Hi! I am doing qRT-PCR of GLUT4 in mice liver and skeletal muscles tissue in normal condition. The data showed the similar expression of GLUT4 in both tissue. Any one can explain that in which condition GLUT4 is expressed in Liver or in both tissue? Thanks
Staining 10um cross sections with PAS and immunofluorescent (for fiber type, cell membrane). Need assistance with imaging/ analysis.

We have tried more typical housekeeping genes (GAPDH, HPRT1, b-actin), but these seem to quite variable. Has anyone had luck with any other housekeeping genes in this tissue? Thanks!
Hi,
I am working on developing an MRI based imaging method for measuring/detecting creatine recovery rate in creatine kinase (CK) reaction post plantar flexion exercise.
It's 2D variant, with single slice coverage of 10-20 mm thickness, has already been developed. Whereas I am developing it's 3D varaint with
with multi slice coverage (4-5 mm thick slices), while still keeping the same temporal resolution of 30 seconds. So, using my method I can probe multiple slice. This allow us sufficient resolution in plane (~1mm) and also along the muscle length (4-5 mm resolution with ~30 mm coverage).
Since only 3D method can probe the intramuscular heterogeneity in CK kinetic along the muscle length, I am very interested in knowing some skeletal muscle disorders where the assessment of intramuscular heterogeneity along the muscle length may be valuable. This will make my work, allowing multi-slice coverage, more significant. I would very much appreciate some inputs.
One of such disorders could be Duchenne muscular dystrophy.
Regards,
Dushyant
I have been considering to use either lentiviral or piggybac with Tet-On 3G promoter to overexpress MyoD1 in iPSC lines. I am pretty new in this stem cell research and I came to know from my former colleague that these iPSC lines are difficult to transfect and through electroporation she managed to transfect 30% with simple GFP marker of cells but it was nearly 10-15% when she tried with CRISPR vector. Does anyone enlighten me with my query that which system would be better to go for. Piggybac is simple but it depends on transfection whereas lenti should be more effective with integrating since cells will not need transfection but infection. But virus infection is how much effective for iPSCs??
I used B-actin on my gastrocnemius sample but it doens't seem to work. The ponceau stain on my membrane showed pretty equal bands, but B-actin on the membranes showed very unequal bands. Any recommendation on a loading control good for skeletal muscle tissue?
Hi everyone
I am working on a 7 segment model of the human body to simulate a walking gait cycle (3 segments for each leg and 1 segment for upper limb).
I have a problem with this model. The order of knee and ankle joints follows the reported orders in standard data (less than 150 N.m in each one of the joints). But the order of Hip joint in each leg is much more than what I have expected. This torque value is about 600-700 N.m.
I wanted to know has anyone faced with the same problem and how should to resolve it?
Regards.
We have a clinical setting, so getting a biopsy of the heart isn't always easy. we are interested in understanding mitochondrial function in the heart in HF. Can I perhaps use blood cells to measure mitochondrial parameters in the heart? Can I use something else instead?
Hello everyone.
I'm using the NucleoSpin TriPrep kit from MN to extract RNA, DNA and proteins from human skeletal muscle biopsies.
Problem is, I usually obtain very low A260/230 ratio (<1.0 and sometimes <0.5), especially when the quantity of RNA or DNA I'm getting from the biopsy is very small (less than 20 ng total).
I tried ethanol purification with sodium acetate and glycogen but I keep getting very low A260/230 ratio, even though I'm being careful with the washing and dry steps. Worse, I'm losing 30% of my RNA/DNA each time I'm purifying them.
I don't have that problem with larger RNA quantities.
Is it my RNA / DNA solution that is still contaminated after one or even two purification, or is it the spectrophotometer that is not sensitive enough for small quantities of nucleic acids and that is giving me wrong results?
Should I use another protocol to improve their quality?
Thank you very much!
Dear all,
I want to perform a KI67 staining on 3D tissue-engineered skeletal muscle of approximately 2cm long and 3mm thick (cilindrical shape). Does anyone already performed a 3D staining with KI67 on tissue with comparable dimensions and if so can I have some tips or working protocol?
I would combine the KI67 staining with a clearing as well (this clearing has already been established in the lab).
Thanking you in advance
Kind regards
Lisanne
I have tried the Ogilvie protocol with prestain buffer at pH 4.3 and 4.5 but the fibers are either all light blue or dark blue respectively. I've come across many papers which claim to use this protocol with mouse tissue and stain all 4 fiber-types, but I don't have any other colors but blue. Am I dehydrating for too long? Can anyone help? I'm interested in changes in the ratios of type II fiber subtypes between experimental conditions. Thanks!
I am trying to isolate RNA from mouse skeletal muscle but I am not able to get a good 260/230 reading. I am using Qiagen RNeasy kit. I would appreciate any suggestion on isolating pure RNA from mouse muscle tissues.
My 260/280 reading are 1.98 but 260/230 is very low.
Hello,
I am wanting to express mouse skeletal muscle contractile performance to lean tissue mass, and I had been advised in a piece of peer review for a paper that it is possible to perform such an analyses by placing skeletal muscles into chloroform methanol for several days, drying, and expressing contractile performance to the remaining lean tissue mass. I am struggling to find anything in the literature to have used this approach for extracting lipids from mouse skeletal muscles.
Is there a published protocol regarding this analyses, or does anyone have a step-by-step guide they could direct me to? Or is it really as simple as the above procedure?
Hi all,
I have been having issues with my samples not running cleanly during SDS PAGE. The samples come from fractionated skeletal muscle where the 'nuclear' samples are causing the issue whilst the 'cytosolic' samples are running cleanly through the gel. I have been using the Bio-Rad Mini TGX pre-cast gels and making fresh 1X TGS buffer running the samples slowly at 80-100V.
My initial thoughts is that it is the sample preparation as this is only occurring in the 'nuclear' samples where the 'cytosolic' samples are being run in the same tank.
Any advice would be much appreciated.
I search a protocol to extract Dna and rna at the same time from skeletal muscle. There are some kit like qiagen allprep but it seem that it doesn't work with skeletal muscle. Anyone have tried a kit for skeletal muscle or a specific protocol?
We would like to test different AAV serotypes to transduce skeletal muscle cells from rat. In order to spare AAV we would like to make some test on adult cultured cells. FDB is easy to dissociate and allows the primary culture of fully differentiated skeletal muscle cells for several days. However litterature on about the use of AAV in these conditions is lacking.
I read a paper about treating muscular dystrophy patient with AAV9 virus infection as a carrier.
AAV9 is known to recognize muscle specifically.
Is there any mechanism that figured out how does AAV9 infect muscle??
Thanks.
Hello,
I have a problem to find primary references showing what is happening with glucose after glucose load (postprandially, during glucose tolerance test, in hyperinsulinemic euglycemic clamp,..). It is usually assumed that insulin drives most of the glucose to skeletal muscle, but I cannot find any references really showing the comparison of individual tissues... Some references to PET studies or other radiotracer uptake studies would be really helpful.
Thank you all.
Ram
I am looking for studies comparing measures of skeletal muscle blood flow using CEU and gold standard imaging techniques in humans