Antibody coating

[Guest Emma]

Hello,

I am using two of the MabTech ELISA development kits (IL-6 and IFNa HRP), following the instructions provided which have been working well. I'm tight on time this week and want to know if it's possible to change the overnight antibody coating step to save time - I've read that with some assays you can incubate at 37C for 2 hours instead. Please can someone let me know as soon as possible if this would be suitable to do with the kits? Or what the minimum time for coating at 4C is?

Thanks! Emma

[Christian@mabtech.com]

Hi Emma,

It certainly works doing a quicker coating protocol of 2h in 37C. However, we have never done proper studies at Mabtech comparing our standard protocol (4C overnight) with that of 2h in an incubator. It is possible that the sensitivity of the assay would go down slighly, but that is a pure speculation. My gut feeling is that the difference is very very small. 

But this you will have to validate yourself. 

[Guest Emma]

Hi Christian, that’s very helpful, thanks for your quick response! Emma

[Lena@mabtech.com]

Hi Emma,
Yes, I agree with Christian and it is possible to adjust the protocol a bit (but as Christian writes we have not done comparison studies). I sometimes coat in RT (for 3hrs) and block overnight instead, if that suits my experiment schedule better.

If you are tight on time, you could instead go for the ELISAPRO kits, with precoated plates. That will save you both time and decrease variability. 

Link to ELISAPRO IL6: https://www.mabtech.com/products/human-il-6-elisa-pro-kit_3460-1hp-2 
Link to ELISAPRO IFNalpha: https://www.mabtech.com/products/human-ifn-alpha-elisa-pro-kit_3425-1hp-2

Good luck!

Lena