I want to detect BRCA1 protein in goat milk with ELISA. But I can't be sure that I use milk or milk whey... most of the kits user manual say there is no problem in milk samples. Producers and firms say there is no problem with milk but they aren't sure. I couldn't find any study about BRCA1 detection with ELISA. The kits fee is very much so I must be sure. Thanks for helps...
Hi Huseyin,
I think it will be OK to detect your protein in goat milk by ELISA. In my experience, serum protein will not affect the detected efficacy in ELISA even protect your protein by increasing its stability. But make a series dilution of your sample maybe a good choice to avoid the interference effect which you are concern.
Good luck.
Weber
Hi Weber,
Thank you for your helps and suggestions. I see your skills and expertise and want to learn more about ELISA. May I ask a few question to you?
1. I decide to look some protein which translated from its gene. I detect the genes in milk from somatic cells. So, normally I wait to see the proteins in the milk. But this proteins molecular weights are not much and it is possible to say there is no much these proteins in the milk. I hope the proteins can be detect in whey with trying some dilutions. Some scientists study milk for protein detection with ultra high speed centrifuge like 44.000 g. When I look the ELISA kits user manual, I see 2000 g or 3000 rpm. I think I do it with kit suggestion but I am not sure. Do you think it is a problem or not?
Thank you again...
Huseyin
Hi Huseyin,
First, it's not represented that the protein you are interested can must be expressed despite you detect genes from somatic cells. In some cases, there were low protein level with high mRNA level.
Maybe you can dilute your standard sample with the same buffer as your sample (e.g. milk...) to rule out the interference effect of diluted buffer.
I'm not sure about the purpose of the centrifuge step, is it trying to increase the collision frequency between protein and capture Ab? If that is correct, you can just follow the kit.
Best,
Weber
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