Questions related to Cholesterol
standard diet for normal groups, and high-fat diet for the rest groups. From the references I read, the normal cholesterol for rats was around 50-100 mg/dL, but the normal rats without any high-fat diet given have cholesterol level above 100 mg/dL. Does anyone have reference about the cholesterol and hypercholesterol value level for rats?
I am studying the interaction of a peptide with components of a membrane. One of the components, for example, cholesterol, is present throughout the membrane. What I want to do is to calculate the distance between the center of a particular amino acid like Phe (at 4th position) and center of the Chol molecule that is present closest to the amino acid.
I used gmx make_ndx to isolate indices of the aa residue and all the chol molecules, however, the indices of the cholesterol molecules are all clustered together without any separation in the text file, hence when I use gmx distance with the select flag, I get the center of mass of the whole system of cholesterol molecules and not the COM of each molecule. This is the command line I used: (I renamed the groups)
gmx distance -f md_trial2.xtc -s md_trial2.gro -n 4PHE_With_Headgroups.ndx -oav avgdist_4PHE.xvg -select 'com of group "4PHE" plus com of group "Chol"'
This works but of course gives me the wrong result.
One option that I have is to manually create index groups for EACH chol molecule and then calculate distances for each of them using a similar command, but there are 96 cholesterol molecules so to do it manually every time would be a daunting task, hence it would be of a huge help to me if you could help streamline this process.
Also, I mentioned Chol as a representative example; I am mainly focusing on headgroups of gangliosides in the membrane.
Thanking you for your help.
Just curious if measuring glycemic and cholesterol parameters in mice after fasting them for 6 hours gives more accurate read -outs than fed state.
I am trying to adapt the protocol found below, but they use THP1 cells, I am using murine BMDM. I am wondering if this is possible? I cannot find purified mouse HDL or apolipoprotein anywhere and we would like to avoid purifying it ourselves if possible.
Cholesterol Efflux protocol I am trying to adapt:
To assess NBD-cholesterol efflux, macrophages were incubated in phenol red-free RPMI 1640 medium containing 5 µmol/l NBD-cholesterol for 4 h at 37°C. Following incubation, the cells were washed with PBS three times and were then incubated with HDL or apoA-1, as lipid acceptors. To determine the correlation, various concentrations of HDL (5, 20, 50 and 100 µg/ml) and apoA-1 (10, 20, 50 and 100 µg/ml) were used for the [3H]-cholesterol and NBD-cholesterol efflux assays. HDL concentrations ranged between 5 and 100 µg/ml, and apoA-1 ranged between 10 and 100 µg/ml. Subsequently, the cells were harvested after 4 h, and the medium and cell lysate were collected for the detection of FI. Triton X-100 (0.1%) was used to lyse the cells in a 12-well plate, and the cells lysate was homogenized by pipetting up and down several times. A total volume of 600 µl was then aliquoted into three wells of a 96-well plate for the measurement of FI. The percentage of NBD-cholesterol efflux was calculated by dividing the FI in the medium by the sum of the FI in the medium and cell lysate. All data were from three independent experiments, each performed in triplicate.
I´m currently working on testing the effects of certain food extracts and their effect on the in vitro micellar solubility of cholesterol. I've been trying to emulsify the cholesterol with taurocholic acid but the cholesterol doesn't even interact with it. I'm using this reactive (https://www.sigmaaldrich.com/MX/es/product/sigma/c8667). I'm thinking about changing to water-soluble cholesterol, but I want to hear your experiences working in this assay or with cholesterol in general.
Are you aware of any good European lab focused on lipidomics of metabolites of Cholesterol synthesis pathway?
Thanks a lot in advance!
Health impacts of saturated fat seems to be contentious. Reports showing negative effects are numerous but inferences of some reports showing positive effects cannot be neglected at all. Based upon latest findings and summing up them with past research conclusions, what can be the best suggestion for common people today ?
I am planning to make cholesterol assimilation properties of my pure cultures. However, I have a water-insoluble cholesterol. So, what is the best medium to dissolve my cholesterol. It is important because, I am desired to use this cholesterol to add MRS broth medium. So, for example, if I dissolved my cholesterol in ethanol, it may have negative effect on microbial growing. So, I need a medium for my cholesterol that have no harm to my microorganisms also.
Whether the pH of lysis solutions (while RNA extraction) need to be modified as per the cholesterol content in the cell's plasma membrane?
Thanks to anyone how would like to spare me attention first. I am working on treating primary mouse hepatocyte with drugs may cause dyslipidemia, hoping to find any positive results on their influences. I have a master question as, what kind of treating medium should I choose? Currently I am applying William E with CM 4000 supplement for both maintaining and treating, while I feel like that too much supplements may have unexpected influence on the outcomes. However, I also wonder that applying poor medium like William E only would strongly damage primary hepatocyte functioning, which may also cause unwished results. May anyone with experiences help me with this?
Besides, I also need a positive control to validate both the SREBP pathway upregulation and cholesterol increasing. Statins can upregulated the SREBPs and Hmgcr by blocking their activity, causing cholesterol lowering, which is not ideal for me. May anyone also have ideas about this can help?
I am treating cells with water-soluble cholesterol. (https://www.sigmaaldrich.com/SG/en/product/sigma/c4951) Most papers recommend 1-10mM. Upon treatment of cells in medium, even within 10 minutes, I can see crystallization of the cholesterol.
How should I treat my cells with the cholesterol?
As increase in level of cholesterol in the body is one of the major factor responsible for heart attack, because deposition of which block the arteries and cause the obstruction in the blood flow. As cinnamon powder has several medicinal values but is any scientific report showing the reduction of cholesterol in the body ?
I'm using methyl-β-cyclodextrin (MBCD) to deplete cholesterol from mammalian cells. I'm following the Mahammad and Parmyrd protocol, in which cells are treated with different MBCD concentrations, ranging from 0.5 mM to 2.5 mM to evaluate the level of cholesterol depletion. However, in the protocol description, they use 20 million cells in one ml, and add one ml of the desired concentration (0.5 mM to 2.5 mM of MBCD). Although the protocol advises not to deplete cholesterol in adherent culture, sometimes is needed depending on the experiment, as I have seen many authors do, but, the reported concentration in those papers is usually 5mM in adherent cultures as if they put a 5mM MBCD solution in medium in the plate or well regardless the cell number. Should I normalize to the cell number in my well or plate? or just pour a 5mM solution on the plate without considering the number of cells?
Thanks in advance
Dyslipidaemia is one of the well established modifiable cardiovascular disease (CVD) risk factors. Elevated levels of total cholesterol, low-density lipoprotein cholesterol (LDL-C) and to a lesser extent triglycerides are associated with an increased risk of atherosclerotic CVD and its clinical end points, Coronary Heart Disease, Cerebrovascular Disease and Peripheral Arterial Disease. In contrary, Elevated levels of high-density lipoprotein cholesterol (HDL-C) are inversely correlated with CVD risk. This is attributed to the anti-atherogenic role of HDL by removing cholesterol from the tissues to the liver "Reverse cholesterol transport" for further metabolism and excretion via bile. What are the possible mechanisms to augment the anti-atherogenic effect of HDL?
I have Martini coarse-grained model of protein in complex with cholesterol. I have followed the backmapping tutorial (http://md.chem.rug.nl/index.php/tutorials-general-introduction-gmx5/others-gmx5#Backward). Now, I am trying to backmap my system (protein-cholesterol), it does not work. It works for proteins only.
I am using the charm36 forcefield, and the initram-v5 script for backmapping.
For cholesterol it says:
Thanks in advance
One of my relative admitted in hospital. he had angioplasty. Interventional Cardiologist recommended him to take medicine Ramipril BP (Controls Blood pressure) and Rosuvastatin (Controls cholesterol) for his entire life. My question is If a cardiac patient control the daily diet (lowers intake of salt and avoid cholesterol containing food) is it still necessary to use Ramipril BP and Rosuvastatin? Thanks in advance
Angioplasty is common problem today. If it is possible to deliver efficient drugs to the exact target, it may be a new era in cardiovascular field of treatment...
We read that people who breathe a lot of dust or asbestos can suffer from fibrosis in their lungs. I have seen that some people also suffer COPD without that and wondered if it has ever been related to high cholesterol? ie. Can fibrosis of the lungs be created from multiple tiny pulmonary embolisms over time when someone has high cholesterol?
I want better repeatability of my protocol and this is hindered by the fact I have to weight each phospholipids everytime i make a new batch which leads to slight changes and errors. So I would prefer to take out those lipids out of a solution instead. I will use the lipids dissolved in ethanol anyway so it would be really convenient.
Can anyone help me with measuring cholesterol and triglyceride levels in zebrafish embryos?
Is there any specific end point measuring kit available?
Experts have also found that restricting food intake during the day can help prevent health problems such as high cholesterol, heart disease and obesity, as well as improve mental health and well-being. Apr 20, 2020 (SOURCE: AL JAZEERA).
I just came across premade NGM (Nematode Growth Medium). It is a powder mix but has Cholesterol to be dissolved with water, then autoclave. My lab always uses the original recipe. We always add Cholesterol after autoclave.
Is Cholesterol autoclavable?
The powder formula I saw is:
Agar 17.5 g/l Sodium Chloride 3.0 g/l Peptone 2.5 g/l Cholesterol 0.005 g/l
I try to analysis cholesterol concentration in serum and liver recently , I see protocol and it says that the serum lipid extract sovlent do not need to add triton, but liver do.
I am curiously about why there are different?
Also, l analysis triglycerides concentration in serum and liver, both do not need to add triton.
l can’t understand why triglycerides do not need to add triton?
sorry for my poor grammar, i hope you guy s can understand what i mean 😢
can somebody help me concerning the preparation of a cholesterol solution for cell culture ? Do I have to prepare a stock solution in ethanol and then dilute it in culture medium at a ratio of 1:1000 ? Any other ideas or possibilities ?
Thanks in advance. Tony
We are planning to add cholesterol to cells in order to raise the intracellular cholesterol concentration and confirm the protein expression of SREBP2. I didn't know if I should dissolve cholesterol in a water-soluble solution, so I asked a question. I would appreciate it if you could teach me how to dissolve cholesterol.
Hello. I have a dataset containing three groups: low cholesterol, medium cholesterol and high cholesterol. Each group has about twenty participants. For each participant I have their mean nocturnal blood gluocse level for 4-6 different days. I would ideally like to compare mean nocturnal blood glucose levels between the three different cholesterol groups. I'm aware that unless I take a mean nocturnal blood glucose across the 4-6 days for each participant and then compare these values then using a one way ANOVA would violate the assumption of independant samples, but doing this would lose a lot of useful data. Is there a way that I can include all of the participant's individuals nights in a comparison between the cholesterol groups? Many thanks
So i have this intriguing idea why dont we make hdl and inject to the patients not in a regular basis, but it will be just as dialysis for cholesterol. Of course it will be taking iv, but actually taking iv saline is like taking water in egypt so why dont we make it more benificial and add hdl to it
I am trying to use water-soluble cholesterol (Sigma C4951) in cell culture. Does anyone know whether I can store this item after dissolving in water, or does it need to be made fresh every time? The product sheet only says that the powder needs to be stored at -20.
I did a liposomal preparation of DPPC:CHOL 1:1 with flow ratio 2:1 (PBS: Ethanol) based on literature of review using microfluidics nano-assembler bench top. I meaured the size using Malvern zetasizer and it was 100nm ( which was expected) however I got very high zeta potential for triplicates (-7330, -7840, -8040mv) any ideas why this happened. size is good but zeta is extremely high.
I repeated the experiment, changed the concentration, checked for contamination. filteration ...etc though it's the same.
I am trying to simulate electroporation for a bilayer containing 10% cholesterol on GROMACS. I have read lots of articles but the only parameter that they have talked about was the electric field magnitude. So I am not sure about the other parameters like angular frequency, time at of the peak in the field strength and width of the pulse.
Is there any references that could help me with that?
l've got rat serum samples with different background (from pink to yellow) and l'm trying to test total cholesterol, HDL and triglyceride in mice serum. l'm worried that the background noise would still exist even l diluted samples. Are there any alternatives to solve this problem? Can l spin serum again (10min, 4c, 1500rcf) before testing? Glad to hear your suggestions!
In a physiology textbook treatment of reproduction, they describe the Corpus Luteum as yellow as a result of the color of cholesterol. Yet, pure cholesterol is white, and has no structures that would lend themselves to color in the visible spectrum. Also, cholesterol deposits in the body, not to mention egg yolks, are clearly yellow. So what is the basis of the yellow color?
I have done electrostatic potential analysis on a symmetric bilayer contains 10% Cholesterol but I've got this figure and I thought there is a problem with that as the potential is not symmetric and it is not equal in the both leaflets.
I will be thankful if you could help me solve this problem.
When walnuts is good for reducing LDL and total cholestrol levels in blood,I think we can boil folium layer in kernel of Walnuts in water and use it as a usefull Liquid.
Sulfa drug increase the the cholesterol and low density lipoprotein.. What is the mechanism of action of this drug which increase these types of lipid...
can anybody help me with In-vivo screening strategy for microbes having Bsh activity? I want to explore them for cholesterol lowering activity?
I'm new to GROMACS and I want to simulate a DMPC bilayer contains 8 Cholesterol molecules. But I have no idea about how to add Cholesterol molecules to my simulation box.
I have determined part of (A) of a two part question where (A): the amount of cholesterol in your sample by reference to the standard curve, which is given to us. I am now unable to determine part (B) of said question which states I should:
Convert that (A) into the total cholesterol content of your 1g sample. CAREFUL you will need to take into account the fraction of the whole sample that you have loaded onto the HPLC.
Hi Dear friend,
Along as I am working on an animal model study as C57lb/6 mice to determining the cholesterol concentration in blood serum and in the brain
But now I am planning to extract the brain and the purpose wants to measure the cholesterol concentration. and if anybody know the procedure please recommend me or give me some idea about it?
I am struggling with visualisation of possible deposition of cholesterol crystals in tissues of gastrointestinal tract.
I am wondering what method tu use, as there is a staining with filipin but from what I undestood - this will stain ALL OF FREE cholesterol (so also this deposited in cells), and I am not sure if that will make crystals vivible.
Second option - is using simply polarized light microscope, and here I would like to ask if anyone has experience or protocol he could share?
I need to check the amount of glucose and cholesterol in zebrafish blood. I have standard kit's from Sigma. My question is, schould I dilut the blood before or not. There is nothing in protocol abouth how to prepare zebrafish blood.
I am searching for a method of visualisation/staining protocol for cholesterol crystals (not a free cholesterol, just crystalized) in cells as well as tissues?
anyone has any experience??
Three week ago, bmj rapid response team publish announcement (Link: https://www.bmj.com/content/368/bmj.m1182/rr-10) This rapid response "as you see" tell us statin like cholesterol lowering drugs may be worsen covid 19 infection. In this research area's (cytokine level, infection rate and statin molecular pathways or etc..) literature very poor. So in this situation born few questions:
1. We use statins reduce cytokine levels. Most of covid 19 case had a severe cytokine storm. So, cut off patient's statins make higher rate of covid 19 infection mortality?
2. A lot of research show high risk for infection people who have a low cholesterol level.
What are your thoughts? Anyone have a molecular and clinical link for statin and covid19?
The following are the reasons for why the High Fat diet does not increases the cholesterol-
- Marked reduction in cholesterol absorption - the mechanism for this effect is not known
- Greatly increased synthesis of bile acids - the patient synthesized roughly twice the mass of bile acids as control subjects
- Reduced endogenous cholestrol synthesis (Kern F: Normal plasma cholesterol in an 88-year-old who eats 25 eggs a day. New Eng J Med 324:986, 1991.).
An increase in the amount of cholesterol ingested each day increases the plasma concentration slightly. However, when cholesterol is ingested, the rising concentration of cholesterol inhibits the most essential enzyme for endogenous synthesis of cholesterol, thus providing an intrinsic feedback control system to prevent an excessive increase. As a result the plasma conc. usually not altered more than ±15 percent by altering the amount of cholesterol in the diet, although the response of individuals differs markedly. A high saturated fat diet increases blood cholesterol conc. 15 to 25 %. Ingestion of diet containing highly unsaturated fatty acids usually depresses the blood cholesterol conc. a slight to moderate amount (Guyton’s physiology).
The study conducted by R B Nair et al, revealed that even after 3 hour of consumption of oil/ghee, lipid profile was not increased, instead there was a decrease in lipid levels after 6 days of Snehana therapy.
Vasant et.al, (Jamnagar, 2006) in their study on standardization of Snehana observed that in group A, triglyceride & VLDL levels were increased significantly after Virecana may be due to Avara Śuddhi. In group B, triglyceride & VLDL levels were increased significantly after Snehana but after Virecana, both have came to normal level, due to the proper Śodhana. So from these evidences, it can be concluded that, after proper Snehana, one has to do proper Śodhana, otherwise lipid levels may be raised and in case of known subjects of obesity & hyperlipidemia, one should have to perform Rukshaṇa Karma before Snehana to avoid the increase of lipids.
Even though the above data shows the safety of use of Snehana therapy, one has to do multi-centered trials on large number of patients with scientific research methodology. Then only myth about Snehana can be cleared from the modern fat conscious world.
Does higher proportion of cholesterol (3%) in high fat diet is affects the feeding behavior of Sprague dawley Rats? Thanks in advance ?
we did plasma measurement of cho and tri. in our mouse samples, now we would like to measure direct level in the fresh tissues, does anyone know where we could do this around montreal area, or any kits that can be used for this!
I am performing a screen to evaluate the role of lipoproteins/cholesterol on gene expression, in which using lipid reduced/free FBS is essential. I have previously used Hyclone Lipid Reduced FBS SH30855.02 but unfortunately Thermo Fisher has discontinued this item. Any suggestions on substitutes from a reliable source? Thank you!
After going through the literature only virgin olive oil looks like help to increase HDL. However, olive contained high amount of omega-9 than omega-3 which may induce inflammation. Despite much hype, there is no good literature to suggest that coconut oil is good for increasing HDL. It increases cholesterol level significantly due to high saturated fatty acids. Flaxseed oil too fail to increase HDL.
I am working on Mice and want to make hypercholesterolemia. But after giving cholesterol powder the level of cholesterol has decreased from previous level. Could anyone please suggest me how will I make hypercholesterolemia in Mice rather giving cholesterol powder?
How do I select only the phosphate group from my phospholipids and oxygen from cholesterol and then club all the phosphates and oxygens into one group in an index file? Kindly help.
I need to calculate the concentration of Total cholesterol in rat tissue samples. can anyone provide me a colorimetric assay without using any kits
i plan to measure the cholesterol concentration in the culture medium to see how much cholesterol is taken up by the cell. But the medium has Phenol Red and it may affects the result? i wonder how to solve that, maybe i can set a blanket as the base? but i don't know how to calculate. the analysis machine i use is Qubit® 2.0 Fluorometer.
Is there a relationship between the serum cholesterol level in poultry and the proportion of cholesterol in poultry meat?
I am novice in free energy calculation with metadynamics. I tried to calculate the free energy (using the attached plumed.dat file) of a biolocal multicomplex system containing lipids, cholesterol, and protein in the form of monolayers separated by a layer of water. The whole system is placed in a priodic box and a NP is placed above the monolayer. The NP freely moves towards the monolayer during the simulation with plumed. Keeping the similarities with the following paper
I wanna calculate the perpendicular distance between the NP and the monolayer surface. Each monolayer contains
What will be the approximate density contour, width of Gausian, height of the Gausian?
I have fine-tuned the plumed input parameters (please see plumed.dat) e.g., the Gausian width, height, and Baisfactor. After 10 ns run with the NP freely interact with the monolayer lipids-proteins, the simulation suddenly stopped with an error (please see Error.txt). I am simulating the system using Martini force field.
Please suggest me about the things that I need in these situation.
Thanks in advace.
I would like to stain slides prepared from biopsies of GI tract for cholesterol (and cholesterol crystals), would you recommend any filipin staining protocol ?
I am looking for a fluorescent probe or stain that would only bind surface cholesterol and that it is suitable for sections of fixed tissue.
I think filipin can be used cells, but I am not sure if it works with tissue.
A protocol would be appreciated.
I've got an intractable issue with loss (apparently) of sterol acetates (cholesterol acetate, stigmasterol acetate, etc) when trying to quantify them via GC-FID. I routinely get 60-70% of what I should be observing. I can observe this both when acetylating standard sterols or when measuring a directly made-up cholesterol acetate standard. My measurement of my standards isn't a problem, as I see the same 30% deficit (for cholesterol) when comparing an acetylated sterol standard with a silylated one (i.e. the TMS derivative gives me what I think I should see, the acetate derivative doesn't). This isn't happening to n-alkyl alcohols, and it's not dependent on retention time (later eluting, non-sterols like archaeol acetate show up just fine), suggesting it's not an injection discrimination problem.
I see a baseline drop after my sterol acetates peak, which does not occur with underivatized sterols or TMS-ethers, or any other class of OH-acetate derivative. The shape of the baseline around the peak would seem to indicate decomposition of the compound on the GC column. However, the problem persists when a brand new column is used, or when a different phase is used (we have used both VF-17 and DB-5 columns, mid-polarity and non-polar, respectively), and there's no obvious reason why these compounds, which are generally quite stable, would be decomposing mid-flight. We've tried reducing the max oven temperature, changing flow rates, varying inlet temperatures, two different GCs, etc. Still keep getting ~70% response. We've tried this with both a PTV inlet and a SSL inlet. We're injecting in toluene. I added a retention gap to reduce interaction with the phase while the compound was 'waiting' to volatilize in the temp program, but this didn't change anything.
Any thoughts? Has anyone ever run into something like this? Or know why an unsaturated sterol ester would decompose on a GC column? I assumed there could be reactions with silanol groups, but the consistency of that that 70% regardless of column age or run parameters is extremely vexing.
I know there are corrections that can be made for the differential FID response of different types of molecule (in this case, a sterol acetate vs my n-alkane response factor standard) for the MOST precise quantification, and I've used simple mass balance corrections or carbon number corrections when comparing acetate and TMS derivatives, but I've never had a problem comparing alkanes and esters to a 1st order approximation before, certainly not to the tune of a 30% difference in response. Thoughts?
I am new to GROMACS and I am trying to understand how to properly build and run a simulation of a lipid bilayer. In particular I am trying to build a lipid bilayer made of three kind of lipids: a free fatty acid (lignoceric), a cholesterol and a ceramide (cer2).
I downloaded the structure files and the force field from ATb (GROMOS 54 a7). Then, following the workflow of Justin Lemkul's second tutorial, I built my lipid bilayer system and run the first equilibrium stages. The problem is the following: once I start my unconstrained NPT production, ceramides and fatty acids stay in their position while many cholesterol molecules leave the lipid bilayer and move "inside" the bilayer, so in between the two leaflets. I know that it is possible that some cholesterol molecules can do this thing, but in my system this happens to too many of them (~30/40%). Honestly it seems really unreasonable and that's why I am here: did any of you have the same problem? Is it related to some kind of bad initial structure?
I am looking for a simple and low-cost method of monitoring the cholesterol content in the cell membranes. My initial idea was to stain them with cholera toxin (anti-GM1 ganglioside) to get information about lipid rafts, so indirectly about the cholesterol content.
Do you have any better ideas?
Both PCSK9 inhibitors ans Statins are very powerful LDL-C lowering drugs. To start with, the JUPITER trial reported a 25% increase in NOD with rosuvastatin 20 mg, over a median follow-up of 1.9 years, compared to placebo and since then, several meta-analyses have confirmed a smaller but significant increase with various statins . Several mechanisms have been postulated underlying the derangement in glucose metabolism by statins. There is some evidence for the detrimental effects of statins on both insulin sensitivity and β cell secretion including the effects of increased influx of cholesterol due to inhibition of HMG-CoA-mediated intracellular cholesterol synthesis, inhibition of CoQ 10 and β cell apoptosis. Also one hypothesis states that lower levels of LDL-C can interfere with optimal β cell function. PCSK9 inhibitors are more powerful LDL-C lowering agents, then why only statins have been implicated in causing NOD and most of the studies done with PCSK9 inhibitors show no association. Is it for some commercial gain to make PCSK9 inhibitors, more popular LDL-C lowering agents?
I have seen several cases of low growth on children (girls) with no hormonal problem, but high blood cholesterol. Thinking about the similarity of the chemical structure between cholesterol and vitamin D, could there be some kind of causality?
Lately I find many cases of low growth in girls (3-4 years old) who also have high blood cholesterol. Could there be a relationship? I can't find studies that associate these two factors and the similarity of the chemical structure with vitamin D is what comes to mind.
I'm using 1X 3b-Hydroxy-5-cholestene 5-Cholesten-3b-ol from Himedia for the culture of Helicobacter pylori ATCC49503 strain in Brucella broth, but unable to find any growth even after eight days. People have earlier successfully used 1X cholesterol from Gibco for such type of culture. Both products are cell culture grade. I'm wondering are they two different types of cholesterol and do bacteria prefer a specific form of cholesterol for their growth.
One of the reviewers of our manuscript said the following about the effects of cholesterol efflux on plasma cholesterol levels, but no references were provided:
There are several previous papers analyzing the effect of peripheral cholesterol efflux on plasma cholesterol levels and concluding that since the RCT pathway is very dynamic cholesterol removed via HDL particles is rapidly esterified via LCAT function and further taken up by the liver and this would not affect plasma cholesterol levels. Only in those cases where for instance liver SRBI receptor or LDLr/LRP-1 receptors are dysfunctional there might be slight effect via RCT on plasma total cholesterol.
If anyone has any further information or reference about this, I would highly appreciate it!
Does anyone know if the lipid levels in mice are comparable to humans ? What is the best method to measure LDL/HDL choleterol and TG in mice ? In advance, thank you for your help.
We are measuring total cholesterol levels secreted into the media by cultured astrocytes using the Abcam cholesterol assay kit (ab65390).
Because the number of cells differ between wells, and the hemocytometer is not very accurate, we are trying to normalize the total cholesterol values.
It was suggest that measuring total protein from the cells themselves, and normalizing: total cholesterol/total protein would be a possible idea.
The premise is that the more protein you have, the more cells there are.
Does this make sense?
Are there other suggestions?