HRP conjugation - Ag-HRP conjugation (Sep/07/2005 )

Hello
I am trying to couple p24 antigen of HIV with HRP.

I have tried Periodate coupling but am not getting significant results.I have done the dialysis as mentioned by the periodate coupling protocol but

Could anyone tell me any other suitable conjugation technique/protocol for Ag & HRP coupling so that I am able to proceed with my expt.I do not have Sephadex G25 column in my lab so am unable to use this technique to purify my protein.

Thanks and regards,
VVI

what about biotinylation of your Ag followed by incubation with streptavidin-Hrp?

mike

[quote name='jadefalcon' date='Sep 14 2005, 03:57 PM' post='24713']
what about biotinylation of your Ag followed by incubation with streptavidin-Hrp?

mike


hi mike
thanks for replying.I am sorry to get back to this so late.
I have been trying the biotinylating of my Ag and incubating with SA-HRP.It still gives background and also the OD is not very much significant.
I used known positive & negative samples whose OD using Ab-HRP conjugate was significantly high.
What could be the reason for low insignificant OD?
Any idea how to otherwise go about HRP conjugation?

thanx
vvi