[Histonet] MCP-1
gayle callis
gayle.callis <@t> bresnan.net
Wed Jun 24 13:42:56 CDT 2009
We never mixed Armenian Hamster and Golden Syrian. Do not use the antiGolden Syrian secondary since the Armenian hamster is a different (and difficult!) little critter than Golden Syrian (I learned this from the head scientist at Jackson Immunoresearch).
If you need an anti-Armenian hamster secondary antibody, buy them from Jackson ImmunoResearch. I ran into some major trouble trying to use an antiGolden Syrian secondary with an Armenian hamster monoclonal primary and also had the wrong host IgG (hamster was the word that led to confusion in my early days of IHC). When the anti-Armenian hamster secondary was used, the staining was excellent. You also need to use an Armenian hamster isotype control - these are available from BD Biosciences, do not use Golden Syrian IgG as the negative control.
We frequently buy our Armenian hamster monoclonal primaries conjugated to biotin but you have to use biotinylated Armenian hamster isotype negative control. This eliminates a secondary altogether and we have excellent staining results, both IHC or IFA.
Ray is correct there may be or not be cross reaction between Armenian and Golden Syrian but with antiArmenian Hamster secondaries available, you can eliminate Golden Syrian from the mix entirely.
Good luck
Gayle M. Callis
HTL(ASCP)HT,MT
Bozeman MT
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of koellingr <@t> comcast.net
Sent: Wednesday, June 24, 2009 12:14 PM
To: Annette Featherstone
Cc: histonet <@t> lists.utsouthwestern.edu
Subject: Re: [Histonet] MCP-1
The hamster isotypes are tricky and ill-defined. Some people (and me formerly) use secondary cocktails. Sometimes Armenian and Syrian are interchangeable. Sometimes they are completely not. BD biosciences has a great Hamster Ig chart that you can use for flow (or IHC) reference.
Ray Koelling
PhenoPath Labs
Seattle, WA
----- Original Message -----
From: "Annette Featherstone" <af46 <@t> buffalo.edu>
To: histonet <@t> lists.utsouthwestern.edu
Sent: Wednesday, June 24, 2009 10:53:33 AM GMT -08:00 US/Canada Pacific
Subject: [Histonet] MCP-1
Is anyone working with MCP-1 Armenian hamster, from Biolegend? This is for
FFPE tissue. The secondary is anti-Syrian hamster. Any help would be
appreciated.
Annette Featherstone
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
More information about the Histonet
mailing list