I have only a minimum experience on ELISA tests and I am trying to use eBioscience kits for BAFF (BMS2007INST) and APRIL (BMS2008) and I'm not having good results. I was wondering if someone who uses these kits could give some tips about what can be changed in the recommended protocol.
If not, what can usually be changed in ELISA kit protocols?
The problem I'm having is that I have some really low reads for my samples, below the lowest concentration point in my curve. I am already using non-diluted serum. Besides, this lower part of the curve is not so good also. When I compare the OD values I get with the ones shown in the kit documentation, my values are higher for the high concentration points, and lower for the low concentration. Even for the BAFF kit, in which the standard strips come ready to use.
More information on the kits: http://www.ebioscience.com/human-baff-instant-elisa-kit.htm
http://www.ebioscience.com/human-april-platinum-elisa-kit.htm
I thank you in advance for any help given! :)
Hi, Liana
What is your ELISA procedure？time, temperature? As the same as the kits told you?
It is normal that the OD values may not consistent with kit documentation shows, you need just a good standard curve and normal sample OD values.
Hi, Zorro
I am following the times and temperatures recommended on the kits. The issue is that even when my standard curve is good, the sample OD values are too low. I was wondering if it could be that these kits don't have enough sensibility. Based on the concentrations reported on the literature (using different kits), I should be having higher values.
Briefly, the whole procedure is made at 20-25°C, and the incubations for APRIL are of 2h with biotin, then 1h with streptavidin-HRP, while for BAFF is only 3h incubation.
Thanks!
Hi, Liana
Does the sample concentrations reported or you wished among the standard curve?
If the answer is yes and your srandard curve is normal, I think the sensibility of kits should be OK, just make focus on your samples, is it really same as reported literature? Is there something different ? for example, the protocol , materials or something else.
If the answer is no, your sample concentrations are lower than the lowest point in the curve, you may try new kits with higher sensibility.
I hope this can be helpful.
Hi Catriona
Sorry for the delay in the answer, but, anyway, I didn't solve my problem. Just gave up on that test... Sorry!
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