Questions related to Blood Biochemistry
Good day all
What is the best position to draw blood from the arm?
Is he sitting or lying down or standing, and why?
Is there a difference in the results that appear through that and why?
Thanks in advance
Some researchers have claimed that Hydroxychloroquine (HCQ) can enter cells and make the environment more "alkaline" by soaking up protons. Many publications discuss only 2 nitrogen atoms that can accept a proton, however the molecule possesses 3, and therefore I would be interested to learn about the transport of HCQ, which is in the diacid form in the commercial Sulfate compound, is potentially in a triacid form in the stomach and is known to be predominantly in the diacid form in Blood at pH 7.4.
There is research demonstrating that the acid form is hydrophobic, compared to the neutral compound, which accounts for its much lower activity against Plasmodium, in which the neutral compound is lipophylic.
How many molecules of HCQ per cell have been found in clinical settings, and would this have any real impact on internal pH compared to natural buffers?
Please share the normal haematology, blood biochemistry and Serum mineral profiles of wild carnivore and particularly Leopard will be of much help. Please share the research articles or any other references related to it.
Thanks and Regards
I will be treating heparinized whole blood with a small molecule for various timepoints and then extracting RNA for qPCR.
Do people recommend incubations in tubes or plates/dishes? 5% CO2/37C or only 37C? I am set on the RNA extraction part, but fairly inexperienced with the prior steps, so any hints or tips would be well appreciated.
My clinical study measured the blood biomakers (glucose, insulin, glucagon, GLP-1, GIP, amino acids, etc) at baseline before an intervention meal and at multiple time points after an intervention meal.
We did this measure three times using three different intervention meals at three different days. My main objective is to compare whether there is any difference in the change in blood biomarkers after different intervention meals.
There are several AUC calculation methods to do this, such as the total AUC, incremental AUC (ignore the area under baseline) and net incremental AUC (subtract the area under baseline). How to determine which one to use and what is the rationale?
Hormesis is a concept to explain of adaptation of body to certain dose of Toxic substance. if some condition in exercise like of produce of Free radical, reactive oxygen species, decrease of pH and etc be a toxic condition for body can we called exercise for one of factor of hormesis? what is your idea?
I want to coat my own tubes with a heparin solution, but can't find any evidence anywhere on what amount of heparin powder I need to add to solution so that the tubes are coated appropriately. I've seen units of 10 IU/mL mentioned, but don't know what this translates to in mass units.
Researches claimed that this blood is more efficient for medical use than blood collected from donors. Patients with rare blood types will benefit the most.
- If a person carries a blood group A resists disease, more than the person that carries a blood group B carries.
- A blood type (also called a blood group) is a classification of blood based on the presence and absence of antibodies and also based on the presence or absence of inherited antigenic substances on the surface of red blood cells (RBCs). These antigens may be proteins, carbohydrates, glycoproteins, or glycolipids, depending on the blood group system. Some of these antigens are also present on the surface of other types of cells of various tissues. Several of these red blood cell surface antigens can stem from one allele (or an alternative version of a gene) and collectively form a blood group system. Blood types are inherited and represent contributions from both parents. A total of 35 human blood group systems are now recognized by the International Society of Blood Transfusion (ISBT). The two most important ones are ABO and the RhD antigen; they determine someone's blood type (A, B, AB and O, with +, − or Null denoting RhD status).
I have recently been advised by a microfluidics device manufacturer to add additional EDTA to EDTA-coated microcollection tubes, since mouse blood tends to form micro-clots.
How much EDTA should I add to an estimated 1ml volume of mouse blood? (Keep in mind that the tubes are already pre-coated with EDTA).
I am attempting to isolate neutrophils from human blood samples. From most protocols I have seen, the process requires 4% Dextran and using Histopaque-1077. Is the Dextran necessary? I want to do this Tuesday and do not have time to wait on a reagent to arrive. Also, I have Ficoll-Paque instead of Hisotpaque. Is that a reasonable substitution? They have the same density.
Previous studies have indicates that reactive microglia expresses molecules such as CD 40 which are necessary for antigen presentation. Can these molecules be detected in clinical blood samples or any other proteins specific to microglia which can be detected in blood?
I want to make a stable blood sample. In my study, I want to make a comparative study of many hematology devices and I want to try many blood sample from different sources.
I've seen many protocols, all of them old Patents, I don't know which I should depend on.
I looked for the answer to this question but I couldn't find any satisfactory answer. I think it is reliable with strong coordination ability of Fe3+ in comparison with Fe2+. However, I need a full explanation. I will be glad if anyone can help.
Ferrozine (disodium 3-(2-pyridyl)-5,6-bis(4-phenyl sulphonate)-1,2,4-triazine) is a water-soluble Fe(II) chelator. When chelated to iron, the complex absorbs light at 562 nm (molar absorption coefficient = 27,900). Copper can also be bound by ferrozine, however copper binds more strongly to another chelating agent, neocuproine.
Iron can be released from protein through oxidation with acid-permanganate (1 hours at 60oC). Excess permanganate and released iron can then be reduced with ascorbate (RT for 30 min). Released iron must be kept at a pH of 4.5 – 5.0 for it to remain soluble. Acid-permanganate is an equal mixture of 1.2 M HCl and 0.285 M KMnO4
Given that the Mr of haemoglobin is 68000 and that for serum albumin is 66000, calculate the number of molecules of iron per molecule of protein. Use the known textbook value for the amount of iron in haemoglobin to calculate the purity of your sample.
Hint: a sensible strategy might be to start by constructing a calibration curve for the reaction of ferrozine with iron. Assays will need to use a control protein (non-Fe containing) for reference.
Another hint: A ratio of 1:5:10 would be appropriate for reagent B: acid-permanganate: sample.
Haemoglobin (1.5 mg/ml)
Bovine serum albumin (5 mg/ml)
Ferrous ammonium sulphate (anhydrous?)
1.2 M HCl
4.5 % (w/v) (0.285 M) KMnO4
Reagent B containing: 6.5 mM Ferrozine
13.1 mM neocuproine
2 M ascorbic acid
5 M ammonium acetate.
There are several molecular forms of Cholecystokinin (CCK), of which CCK-8 and CCK-33 are the two frequently used molecular forms in pharmaceutical infusion studies to induce fullness and reduce hunger in human/animals.
However, in dietary studies, which involve participants to consume a test meal, which molecular form of postprandial plasma CCK should we assess? What are the methods available? What are the advantages and disadvantages?
Many dietary studies have not been explicitly stating the molecular form of CCK analysed, although some did mentioned CCK-8, CCK-33 or total CCK being measured.
So how should be make decision around which form of postprandial plasma CCK should be analysed in appetite studies?
In many instances a small amount (5-30 ML.) of blood from the external surface of body is let out. In the process of homeostasis, some chemical parameters might change. I wish to know- what are the changes in the biochemistry of blood that take place eg. pH and after how much time does the compensatory mechanism stop?
I am looking for C57/BL/6 mice (8-10weeks age) blood plasma biochemical parameters, i.e., blood urea, ALK, GOT, GPT, cholesterol, triglycerides, HDL, LDL.
I need these data as control mice data.
Human blood has ph range 7.35 to 7.45. I want to about refractive index of different ph value. kindly suggest me a link or research article where I get refractive index values for different ph blood sample.
Dear sir, Regards.
During preparing the serum i found that the supernatant clear part is gelatinised? and when i repeated with a new batch of blood, same animals showed gelatizisation again? what may be the cause sir? and what may be the condition.
Dear colleagues, I was wondering if there is a relationship between the concentration of proteins such as Albumin, ALP, and ALT on both sides of a cell membrane where secretion occurs? An example would be the ALP protein level increases by 10% in human bold plasma. Will the final product after secretion from the plasma delivery most likely also increase by 10% from its original concentration after a while?
Blood plasma (ALP up 10%) -> secretion process in the membrane -> Final product on the other side of the membrane (ALP up 10%)
I have prepared PRP from Sodium citrate whole blood and have 1M CaCl2 solution, problem is I have limited sample to test this part of the study on. Previous studies have used 20mM but the ratio of PRP to CaCl is not disclosed. Any one have any tips on what concentrations to try. I have 6 samples that I can try but one of those will have to be the whole blood in order to compare.
Can you please provide some good datasets of blood biochemistry and cell count data available for academic use?
Capillary refill time is one of the sign of dehydration and shock. Capillary refill time is widely used by health care workers as part of the rapid cardiopulmonary assessment of critically ill children because it is a marker of increased peripherally vascular resistance. I think that capillary refill time is a vital sign. What is your opinion about this topic?
I want to perform lipid panel tests on 200 mice every other week for about 3 month. if I use ordinary spectrophotometers which work with ordinary cuvettes, it requires a huge amount of reagents. Besides, based on the catalog of the kits we have, it needs to draw at least 700 microlitter of blood to acquire about 400 microlitter of serum. this is impossible because if you draw 700 microlitter of a mouse blood, it will die or at least it suffers from a huge amount of stress.
so, I had an Idea! is it possible to use a nanodrop or other microvolume spectrophotometers for blood biochemistry tests such as lipid panel?
if it's possible, I will be able to perform all the tests on a single mouse with less than 10 microlitter of serum. in addition, based on my calculations, I will consume at least 50 times less of the reagents.
did anybody use nanodrop for such applications? or heard/read about it?
thanks a lot
We are looking for large repositories of blood biochemistry and other clinical data linked to mortality and other events. Any recommendations are highly appreciated.
Besides fasting blood sugar and glycaeted haemoglobin, what is the current guideline for use of fructosamine and glycaeted albumin as biomarkers of hyperglycaemia?
I want procedures for catalse , super oxidedis mutase , para oxinase enzymes for blood sampels???
Please suggest me about "Methemoglobin determination/estimation/mesurement".
I want to estimate Methemoglobin amount present in the blood of rat.
I will induce Methemoglobinemia in a rat by using some chemicals.
Some paper suggest to use Co-oximeter for this purpose. But I did not find any protocol related to this in the detail.
In my lab we normally do cell culture (For karyotyping) from fresh sample, just after collecting in heparinized tubes. Now we are thinking of doing some sample which will come from a longer distance. Its probably takes 10-16 hr to reach in our lab.
Is it possible to do the test with retaining its maximum quality?. If so then, what measures should I need to take during sample transportation?.
So I am asking this question to know about the storage condition that i need to ensure during transportation.
Stimulation with IFN-g or LPS in 15mls conical tubes mounted on a rotator inside an incubator at37'/0%CO2 for 24hr (so the blood continuously moves around within the tube).
I am trying to quantify the expression of an intracellular protein via flow cytometry in monocytes (gating these cells from whole blood analysis via expression of CD14, CD16, CD45 and HLA-DR). so far i have stimulated undiluted whole blood with vary doses of the two stimulants obtaining poor results.
Iwe have a project looking at leukoreduction effects in equine whole blood stored over 5 weeks in CPDA-1. Oxygen tensions are ~200-225 mmHg by week 2 in leukoreduced bags, slowly increase from 60 to ~100 in non0leukoreduced bags. CO2 increases to ~80 mmHg in lekoreduced and stays there by week 1, increase is to about 120 mmHg in non-leukoreduced. I am trying to sort out why?
People! I collected blood samples, separated the plasma and frozen it, but when the samples were transported to my laboratory, they unfrozen.. I don't know how long they remain in this situation (unfrozen). Do you think I can use them to analyse lipids, triglycerides, uric acid, cholesterol, lactate? I can't lose them it's for my mastership!
I am doing targeted protein quantification by mass spectrometry. The interesting protein called frataxin. It is about 14-18 KD protein in mitochondria inner membrane. I heard how to get mitochondria enriched cells or platelets from fresh whole blood, however, my future samples will be frozen whole blood in EDTA tubes. How can I get mitochondria enriched part? Thanks.
I received a blood bag for PBMC isolation with subsequent T-cells triage required. I received it yesterday evening and put the bag at +4C. I wonder if I can do a ficoll etc NOT the next morning but the morning after? So it will rest for 24+ hours at +4C... Is it bad for the experiment?
Thank you in advance!
I would like to know if there is some portable analyzer similar to those that measure lactate, e.g., Lactate Pro.
I'm currently trying to detect the signals of intracellular LC3 in isolated eosinophil from human blood. After being purified (the purity is ~ 99%) and treated, eosinophils are harvested, permeabilized immediately (with 0.1% Triton-X) and incubated with PE-LC3 antibody, and resuspended in 1X HBSS. Recently I got two problems:
1/ Another cell populations shift towards SSC high/FSC high. This makes it harder to gate the general population.
2/ In some samples, signals of PE-LC3 were very high. So I think maybe it's due to unspecific staining.
Is there anyone has encountered these problems? I appreciate any help you may provide.
I have found red cell sodium values (calculated values only unfortunately) to be 'low' - sometimes 'undetectable'. I am guessing that a value around mmol/Litre
I would like to ask if there is calcium really unnecessary for thrombin activity ? I've found one publication, where they claimed that thrombin is Ca-dependent enzyme ... but there was no reference to this statement. I need to use thrombin to cut-off GST from my fusion protein, but my protein-of-interest is vulnerable to Ca and Ca lowest it's activity.
Here is the quote from one paper that mess my mind:
We have found that recombinant TGase loses activity in a matter
of hours in the presence of calcium, so we were concerned that
the calcium-dependent thrombin-mediated cleavage of Lai’s GSThTG2
fusion protein would lead to significant loss of activity of
the resulting hTG2.
For me, the availability of each Amino Acid in fasting blood RELATIVE to all others is more significant metabolically, than the absolute values of each AA (which are currently assessed individually to ascertain whether each falls within a said 'normal range' or not), as:
Consideration of all amino acids together and relative to each other, provides context, as well as strengths and weaknesses of the metabolome as a whole.
I want to measure hematocrit on small samples, but don't have a microhematocrit centrifuge.
Is there a special insert that can be used in a normal centrifuge, or just another method that may be used altogether?
I need to detect some heavy metal concentration from plasma and do some enzyme essays from serum or both. Please suggest me if there is any technique in which both plasma and serum can be harvested safely from the blood.
We have blood contained in paxgene tubes and would like to know whether Trizol method would be effective in isolating RNA. Is there any modification to be made in the trizol method, for isolating RNA from blood contained in Paxgene tubes to obtain maximum yeild and integrity.
Blood samples once collected in paxgene tubes are stable for a long time and are required to be extracted using the paxgene extraction kit. Instead of using the kit, I used trizol method for the same, and got a single faint band(opposed to 3 bands).
Does anybody know if it is possible or have a protocol for measuring citrate synthase activity in serum? I have already measured activity in skeletal muscle and want to see if the changes can be tracked in the blood stream.
Previously I used a protocol using DTNB to measure the reaction between acetyl CoA and oxaloacetic acid.
I have three frozen, centrifuged blood samples in Yellow-cap vacutainer ACD tubes. They are currently being stored at -80C, and I need to extract DNA from them. They fill about 1-2ml of the tube per sample.
If they thaw out, will the buffy coat layer stay separate, or will the layers merge right away?
If the layers merge, I will be able to extract just from the buffy coat, otherwise I will have to extract from whole blood.
I am attempting to quantify a particular protein in red blood cells and I want to remove hemoglobin so that it will not interfere with the assay that I am using.
We have power trips going on quite frequently in our lab and to say the least this is a nightmare for my PBMC isolations. I was wondering if it would ruin the layer separation permanently or is there still a chance to get a nice layer by resuming the spin after electricity restoration? I wanted an opinion from someone who has experienced such an event.
We are using human I-FABP kit to detect in pig plasma samples. I cannot find any literature on abundance of I-FABP in plasma, only in cells. We want to optimize this protocol. We know the total protein concentrations in our samples, but I'm not sure where to go from here.
Any tips, tricks, and general information is most welcome!
I tested some blood serum parameters like HDL, LDL, trigyceride etc. and leptin in albino wistar rats. When I look at the literature I could not find a direct answer about right timing of blood collection. If I applied sample collection in the same way, same time, same row for all animals, can eating of rats before blood collection still create a huge individual variance differences and avoid to have significant data for treatment? Please help me, I am completely confused.
Has anyone ever quantified the relative yield of RNA from whole blood, serum and plasma, preferably from the same original sample?
Thanks in advance,
It is well known the high affinity of clotrimazole for the haeme moiety. I wander whether its binding to haemoglobin has been studied and if it depends on the oxido-reduction state of the Fe atom. I would greatly appreciate any comment.
I need to know about new techniques can use to look for circulating histones in plasma from patients who may be exposed either to trauma or injury, or may be as a result of diseases.
We have been trying to isolate RNA from whole blood using different kit methods (Promega and others) but we get degraded RNA every time. Please tell us any manual method like Trizol and CTAB method in which we do not need to extract buffy coat first. Also indicate the homogenization procedure. Is it true that haemoglobin binds RNA and precipitates with it so that is why we are not getting RNA?
I wanted to get a feedback on the use of infrared or Raman spectroscopy for detection of breast, cervical lung cancers or in detecting blood biochemistry.
Has anybody used the method for detection of markers such as EGF, p16, CEA, CA125 for early cancer detection?
I Want to extract Angiotensin peptide from blood, already i have synthesized biologically active Angiotensin with higher purity, Now i want to extract natural peptide and compare the biological properties of both, Am looking perfect procedure from Experts.,
I've tried an ELISA with mixed results so I would like to confirm the results by western blot. However, serum does not run well on PAGE gels. Any idea if lyophilization and resuspension in water will help? I ran serum on a 10% gel and it was close to the worst gel I've ever run in my life. If anyone has any helpful suggestions, I would appreciate them.
I would like to measure and monitor pentraxin 3 levels in ICU patients. Any experience how to store the serum samples and where? Also I can't find the half life of PTX 3 in the literature any ideas. Thanks.
In my lab I use Olerup-SSP HLA-Typing tray kit for HLA-A,B,DR typing of patient and donor prior to check the match-ability for organ transplant .We use manual (data sheet) to find the serology and allele regarding found bands, so is there any software available to analyze the HLA-Typing ?
One should add aprotinin before centrifugation of samples for plasma or serum isolation and then freeze for subsequent small peptides' determination. If this step has been missed, is there any sense in adding aprotinin after thawing such samples?
It took me half an hour to separate serum from Guinea pig blood by allowing blood to stand still in a slant position at room temperature followed by centrifugation. I need a quicker means of serum separation to produce high-titre complement.
I will like to collect blood for both PCR and ELISA. I want to avoid using two different tubes. Is it possible to do ELISA from blood collected in PAXgene tubes? Or will the preservatives interfere with the assay? I know it is fine for PCR but is it okay for ELISA?
I have read a lot about drawing blood from Wistar rat and I have knowledge about different techniques. For my current research, I need a technique that wouldn't include the death of the animal (e.g decapitation). In the past, I have tried drawing blood from tail (which seems to be the easiest way), but unfortunately I didn't succeed, what so ever. I need help with drawing blood from Wistar rat and I'll be happy to have any NEW methods or any extraordinary tips.
A preliminary demonstrated that I could not find any specific diseases that nitric oxide and S-Nitrosothiols are really good predictors for.
Diseases such as alzheimer's are so big that i doubt providing clinicians with a quantitative skin-nitric oxide/ S-Nitrosothiol level would be relevant.
I am planning to determine the amount of Zinc released and distributed from Zinc oxide Nanoparticles in in vivo conditions in organs and blood of mice. As cost and availability of instrument is a limiting factor, I would like to know techniques other than ICP-AES/ ICP-MS for the analysis.