Sarah,
I am curious; have you tried contacting the BioCare research tech
support reps? They are very helpful regarding their products and
trouble-shooting any issues that may arise.
Akemi Allison BS, HT (ASCP) HTL
Director
Phoenix Lab Consulting
Tele: 408.335.9994
E-Mail: akemiat3...@yahoo.com
On Apr 28, 2011, at 1:16 PM, <sgoe...@mirnarx.com>
<sgoe...@mirnarx.com> wrote:
So I am staining tumors that were implanted as cells subdermally into
mice. The cells are human. I am trying to do Caspase staining on
these
tumors. The primary is an anti-human rabbit polyclonal, and I am
using
a polymer (Biocare Mach3) in lieu of the secondary antibody. The
background is through the roof!! Could the reason be that the
tumor was
grown in a mouse and is having cross reactivity somehow? What species
antibody should I be using instead? All my mouse monoclonal
antibodies
work perfect on the tissue, it's this stupid rabbit polyclonal!! I am
blocking endogenous enzymes (peroxidase etc., DAKO), avidin and biotin
(just to see if that would help...it didn't), and protein block (it's
literally an hour worth of blocking!!), developing with DAB (Dako) and
hematoxylin counterstain. I am so confused as how to get this to
work!
Also, it isn't just this particular antibody it is any rabbit
polyclonal
I have tried. Could it be the polymer? It is the one that Biocare
suggested? HELP!!
Thanks in advance =)
Sarah Goebel, BA, HT(ASCP)
Histotechnologist
Mirna Therapeutics
2150 Woodward Street
Suite 100
Austin, Texas 78744
(512)901-0900 ext. 6912
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