Untested Antibody for IHC - (Feb/02/2011 )
So I'm interested in BRM immunohistochemistry in mice. However, the only antibody that I come across is the one from BD, and it's rated for IHC (wb/icc/and other junk). However, I tried it on one of my western blot and nothing came up! The same extract showed up with another antibody (abcam/santa cruz).There's a possibility that the BD BRM antibody I have at the moment went to hell.
I can't find literature on BRM mouse IHC, I keep running into BRM human IHC though. And looks like I'll have to test several kinds of antibody (saw a paper that claimed to have used Santa Cruz BRM for IHC, but IHC is not rated on Santa Cruz's site).
So my question is, can I use the same protocol for different antibodies. If not, how much modifications are we talking about? Sorry for such a broad, generic question. But without literature, I don't even know where to begin 
Thanks!
if you are asking if you have to change your method then the answer is yes, you can use the same protocol.
if you are asking if you can use them for different purposes (ihc, wb, icc, elisa, etc) then maybe, maybe not, depends on epitope specificity.
Here's a list of a couple of other suppliers. Some have publications too:
http://bit.ly/eUF2bg
mdfenko on Wed Feb 2 19:53:31 2011 said:
if you are asking if you can use them for different purposes (ihc, wb, icc, elisa, etc) then maybe, maybe not, depends on epitope specificity.
Well here's the thing. The BD antibody is rated for IHC, WB, etc. But it didn't show up on my western. That's why I'm hesitant to try IHC with this version.
Then I came across a paper (with very little detail on their IHC), and they claim to have used the Santa Cruz antibody, even though the SC antibody isn't rated for.
That's why I want to test various versions of antibodies, seems like manufacturers aren't very consistent with their testing/certification.
Most antibody companies only list applications that they have tested in house. So it's possible that the products will work for unlisted applications. Also bear in mind, that re-packaging and re-branding is common practice amongst antibody suppliers, so the same antibodies will appear in multiple catalogues with different product numbers.
Antibodymania is correct many suppliers rebrand and repackage one anothers products. You should go ahead and try the ab in your application. I would suggest you screen them using the same protocol and using the abs at the same concentrations.
Say Chi Sin Lo on Wed Feb 2 21:17:27 2011 said:
Well here's the thing. The BD antibody is rated for IHC, WB, etc. But it didn't show up on my western. That's why I'm hesitant to try IHC with this version.
Then I came across a paper (with very little detail on their IHC), and they claim to have used the Santa Cruz antibody, even though the SC antibody isn't rated for.
That's why I want to test various versions of antibodies, seems like manufacturers aren't very consistent with their testing/certification.
does the data sheet give you a recommended dilution for wb? maybe you were too dilute.
how old is the antibody (not just how long you have it)? maybe it is not as active as fresh.
you can find proper dilution by running dot blots or by using a "surf blot" apparatus on a preparative gel transfer.