[Histonet] Mouse CD markers, Histosearch, apoptosis and San Diego

[Donna Harclerode]

Hi Liz

I have been trying to search for a positive control for rat apoptosis
and have not been able to get on the archives either.  It is such a
valuable resource it really hurts when it is not there. 

I never have been able to get my frozen rat hearts to work with either
the Chemicon or Roche kit for TUNEL or many of the apoptosis markers
(Caspase 3, PARP and DFF45).  I have taken the same frozen tissue (after
sectioning it for frozen), thawed it and processed into paraffin and get
Fantastic TUNEL so I am going to try the other markers in paraffin.  Is
anyone doing apoptosis in frozen rat tissues? Is there some tick I
cannot figure out?  The only reason we need frozen is for CD31, which
hopefully the only ab that works may available from Santa Cruz soon.  I
got the available lots of all their available abs. and tried them awhile
back on paraffin and nothing worked.  I have read on the list that they
are expecting a new batch soon, but not yet. 

Mouse CD markers
PharMingen mouse CD11a clone M17/4 works in frozen sections (1ug/ml)
with polyclonal secondary.  The other CD 11a clone 2D7 works at 10ug/ml,
but only with the isotype specific secondary and not the polyclonal.  
The CD54 3E2 works well at 5ug/ml BUT you must use the hamster cocktail
from PharMingen.  (Jackson Immunoresearch may have come up with Syrian
hamster secondaries by now, but I have not used them)

I worked for PharMingen setting up the QC testing for IHC applications
for their mouse, rat and human antibodies back in 1997 so I have
recommended concentrations, tissues used for testing the antibodies
available then and know the ones that need special secondaries.
PharMingen has dropped some recommended applications (like mouse CD31,
MEC13.3 for paraffin), but it worked pretty well if you used trypsin
pretreatment and not microwave. 

The San Diego chapter of the California society has been dark for a
couple years.  After the Christmas holidays I am going to try and get
our group up and running again.  If you are in San Diego area and
interested in for educational meetings, drop me and email and I will be
sure to let you know what is going on.  If you are willing to help get
the group going again please lat me know that too.  I appreciate all
volunteers!

Donna Harclerode, HT, (ASCP), HTL, QIHC
Immunohistochemist
Cytori Therapeutics
6740 Top Gun St.
San Diego, CA 92121
dharclerode <@t> cytoritx.com



Is anyone else having problems searching the archives?  I have not been
able to get the search page in days.  I wanted to check to see if CD11a
and CD54 for mouse tissue will only work on acetone fixed frozen
sections.  I'm sure that this has been brought up before but
unfortunately I can't get to the search page.
 
Thanks in advance.
 
Liz
 
Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Manager
Premier Laboratory, LLC
P.O. Box 18592
Boulder, Colorado 80308
Office: (303) 735-5001
Fax: (303) 735-3540
liz <@t> premierlab.com
www.premierlab.com
 aries etc.

Is anyone else having problems searching the archives?  I have not been
able to get the search page in days.  I wanted to check to see if CD11a
and CD54 for mouse tissue will only work on acetone fixed frozen
sections.  I'm sure that this has been brought up before but
unfortunately I can't get to the search page.
 
Thanks in advance.
 
Liz
 
Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Manager
Premier Laboratory, LLC
P.O. Box 18592
Boulder, Colorado 80308
Office: (303) 735-5001
Fax: (303) 735-3540
liz <@t> premierlab.com
www.premierlab.com