ELISA and BSA - Non-ELISA grade BSA (Jun/28/2006 )

Hi, everyone, i also have a question in my ELISA protocol. can i use BSA as the blocking agent when the coating protein is OVA (ovalbumin), thanks.




thumblina

Try to use 10%FCS/PBS, or you could try to block the plate O.N. at 4 degree.

we use 2-5% normal goat serum to block (we use goat secondary antibodies).

Hi all,
I've tried with different blocking agents and with my ELISA.. Caesin is not good enough.. as mine was dealing with amyloid beta..

in my experience its better if you go for a commerical blocking agent.. I found superblock in TBS from pierce works fine..

good day!!
Saint

hi,I need help from you!!! i'm working with competitve ELISA (detection of antibodies) in human brucellosis. The principal protocl assay don't use BSA or another blocking agent. For that, i tested differents concentrations of BSA (1, 2 and 3%) but my results were identically for the positive and negative controls (no differentation between them) and their optical densities were 0,2-0,4. The protocol also use EDTA/EGTA in PBS/tween solution as sample diluent buffer and i read that this chelators agents are used to reduce interaction non specific with another serum proteins.

Do you think that this could be interfering with BSA? how could it be?
The use of EDTA/EGTA in the diluent buffer is replaced by the used of the BSA?

thanks.

Pilar

question about BSA in ELISA

We use sigma - non-elisa grade BSA for one ELISA and it works fine. The other two ELISA's we do utilise the BD opteia commercial elisa blocking media - works great.