Many companies providing single reagent solution for enzymatic reactions, by which we can quantify cholesterol, triglycerides, and HDL-C. Can I use the these human compatible reagents with rat or mouse sample? Is it okay scientifically? Thanking you in advance.
At molecular level there HDL-C shows differences in molecular weight and shape, It has difference in % cholesterol and hydrocabon percent in its molecule. That effect cholesterol efflux and transport inside the cell. Rest of the molecules will be of size and same formula there will be no difference, that you can use.
There is no significant difference between the two samples. Results are comparable.
You can use these tests for animal samples, the only "problem" exists in the missing reference values in the instruction leaflet. It´s something different if you want to quantify the apolipoproteins because most human ELISA don´t crossreact with rodent samples.
yes, if u mean enzyme based colorimetric/somtimes even fluortuimetric methods.
Agree with the 'reference value' issued raised above. But I assume you will have your control group as well.
Single reagent or one-step analysis could be erroneous for triglycerides since free glycerol will contribute to end analysis. We had to apply a small correction for protocols that use one-step reagent to account for free glycerol. Pls check the paper
http://www.ncbi.nlm.nih.gov/pubmed/23597845
Hi Annadurai, As you know cholesterol including total cholesterol , LDL- Cholesterol and HDL-Cholesterol , have the same structure, formula and chemical reactions from different species. It means Cholesterol oxidase reaction with human cholesterol and rat cholesterol and rabbit cholesterol are the same, the products of the reaction are the same too.
So you can use human cholesterol assay kit for measuring Rat serum cholesterol too. But there is a fine point. This is matrix effect. The matrix contents of human serum are different from Rat or Rabbit matrix serum. It means with Human Cholesterol assay kit you can determine normal cholesterol level and range in human sample. But you can not use human kit for rat normal value determinations.
In human sample, cholesterol e.g. 120 mg/dl will have acceptable precision and accuracy. But using human kit for rat serum, e.g 76 mg/dl cholesterol, may be its accurate amount a small higher or lower than this, for matrix effect.
But in many animal study, we do not use normal value, and we need comparison between case and control group cholesterol amount.
Finally, in comparative study including before-after study and case control study, you can use human cholesterol or TG assay kits for e.g. Rat samples.
But if your mean is normal value determination for rat serum or cut off defining for cholesterol in rat, It is not recommend human kit using for other animal samples. the main reason related to matrix effect.
best
mehdi
You can use such reagents to the measurement of TG,but take care with the glycerol content of the rat sample because free glycerol measurement is also growing when that condition is present whenrats are in an active lipolytic state. LDL and HDL- cholesterol may be run with the same reagents used for human molecules. Take care because data interpretation is different between human and rat plasma samples. For example, rats don't present cholesterol ester transport protein (CETP) , so rat LDL and HDL molecules show different percentage of cholesterol esters in comparison with LDL and HDL from human samples. A control group is always needed.
In my experience, you can use cholesterol and triglycerides reagents, but HDL reagents may not be useful, particularly in hyperlipidemic mice (see Clinical Chemistry 10/1999; 45(9):1567-9).
In my experience, reagents used for human sample analysis work fine with rodent samples