I would like to refresh this question.
Which vacutainer are You using (EDTA or EGTA) and can anyone recommend an elisa kit?
Hi Daniel,
I recently tested the IBL plasma adrenaline kit #IB89558. It includes a solid phase extraction step and acylation followed by an incubation on a microtiter plate with the antibody. Finally, you add the sample-antibody mixture to the EIA plate and proceed. The assay takes the better part of a day to get to the step of loading your preincubated samples on the EIA plate and that incubates overnight, then finish that assay with conjugate and substrate steps the next morning.
Good luck!
Hi Elizabeth,
thank You for your quick reply! I will test it in a few weeks and let You know how it worked.
Hi Daniel,
Glad to help! Some small tips: I did not have the best replicates in my first attempt. Tech support told me that it is best to use a multichannel pipette for the wash steps, but I had used a squirt bottle, which is fine for most EIAs. Also, they tell me that it is advisable to use a repeater to add the enzyme solution to the extracted sample that you have transferred to the microtiter plate.
One other thing. The protocol is written in such a way that they describe the preparation of the enzyme solution at the beginning of the protocol THEN tell you that it is good for 15 minutes. If you prepare the enzyme at the beginning, it will be no good by the time you are ready for it. I recommended to them that they consider changing that in the next protocol revision, but in the meantime, one has to be careful not to follow the directions step by step in order.
Dear Elizabeth Oseid!
I want to ask question regarding my PhD Research. I have seen your answers in a discussion on RG that is why approaching you for expert opinion.
Please tell me that: I have frozen rat plasma samples at -80 C and want to measure glucose levels. Is it possible to measure by a Glucose Meter intended for human use?
Thanks
Dear M Atif Raza Cheema,
You certainly may use a human BG meter to measure BG in rodents, but the sample should be whole blood, as from a tail stick. For plasma, it will be best to use a glucose analyzer, but these are expensive and require a fair amount of maintenance. I use one routinely, because we do some human studies involving hypoglycemia, and the analyzer is more accurate at the extremes, while both are good at normoglycemic levels.
That being said, I have never tried to use plasma with a hand held meter. If I were you, I would do a finger stick on myself, then have someone draw my blood and immediately test it prior to centrifugation, then spin down the blood and test the plasma. Depending on your results, I would do the same with a rat.
As for your existing samples, if your results from the above test are not satisfactory, I woud try to locate someone with a glucose analyzer, possibly in a hospital research center and see if they can either run the samples for you, or teach you to use the analyzer.
Good luck!
Elizabeth
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