Hi Jakub,
Purify your B cells first then use PMA (phorbol
ester) 10 ug/ml and ionomycin 1 ug/ml.
LPS should have worked but its not specific to B
cells.
As for monkey anti-IgM ( anti-mu should be better),
try the antibody resource pages on the web there
are many usually as part of a company site.
You might need to crosslink the anti-mu to get a
mitogenic signal. Even better if you stimulate your
cells with IL-4 and anti-IgM.
cheers
david
In article <382A22EC.D1A9E24F at stanford.edu>,
Jakub Regieli <jregieli at stanford.edu> wrote:
> I'm trying to achieve polyclonal activation(and proliferation) of
> B-lymphocytes in vitro.
> I'm having difficulty finding the right stimulus (mitogen or MAb) to add
> to culture, to stimulate specifically B lymphocytes.
>> I tried Lipopolysaccharide (Sigma E.Coli 055:B5), and Staphylococcus
> Aureus Cowan I (Calbiochem, product name Pansorbin).
> None of these worked, as they should according to literature. I might be
> using the wrong clone or origin of the stimulating agents. I'm working
> with peripheral blood of cynomolgus monkeys.
> I'm not sure where to obtain anti-IgM antibody that would crossreact.
>> Does someone have a suggest where to search for the correct mitogen?
>> I'd appreciate any help/directions you could give me.
>> ------
> Jakub J. Regieli
> Transplantation Immunology
> dept of Cardiothoracic Surgery
> Stanford University
> lab 650 - 498 - 6414
>>
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