src family ptk inhibitor pp2  (Biomol GmbH)

Journal:

Article Title: Epitope-specific crosslinking of CD45 down-regulates membrane-associated tyrosine phosphatase activity and triggers early signalling events in human activated T cells

doi: 10.1111/j.1365-2567.2004.01986.x

Figure Lengend Snippet: TNF-α gene expression requires PTK and PKC activity. Primary activated T cells were preincubated for 30 min with PTK inhibitors genistein (a), tyrphostin (b), herbimycin (c), or PP2 (d), or with the PKC inhibitor calphostin C (e), then simulated for 2 hr with anti-CD45 mAb 4B2 crosslinked on plastic. Similar results were obtained with cells stimulated with mAb 9.4. Alternatively, T cells preincubated with tyrphostin were stimulated with PMA (20 ng/ml) and Ionomycin (1 µm) (f). Results are expressed as TNF-α mRNA fold induction over unstimulated T cells (representative sample in (a)). [32P]-labelled PCR products quantified using an Instant Imager™ are presented in boxes (TNF-α transcripts: upper signal, internal standard transcripts: lower band).

Article Snippet: The Src family PTK inhibitor PP2, PMA and ionomycin were from Calbiochem (La Jolla, CA).

Techniques: Expressing, Activity Assay

Journal:

Article Title: Epitope-specific crosslinking of CD45 down-regulates membrane-associated tyrosine phosphatase activity and triggers early signalling events in human activated T cells

doi: 10.1111/j.1365-2567.2004.01986.x

Figure Lengend Snippet: Tyrosine phosphorylation of PTK ZAP-70 and p72syk, adaptor protein LAT and p23-ζ induced by CD45 engagement is dependent on p56lck kinase activity. Jurkat T cells were preincubated or not with PP2 (10 or 100 µm) for 30 min, and were then stimulated with soluble (S) or crosslinked (XL) anti-CD45 mAbs 9.4 (20 µg/ml) (or 4B2, data not shown) for 15 min, or left unstimulated. Whole cell lysates were immunoprecipitated with specific anti-p72syk, anti-ZAP-70, anti-p56lck, anti-LAT and anti-CD3-ζ antibodies. (a) Following separation on 6·5–10% polyacrylamide gradient gels in SDS and transfer to PVDF membranes, blots were incubated with antiphosphotyrosine mAb and revealed by chemiluminescence. (b) Blots were stripped prior to incubation with antip72syk, anti-ZAP-70, anti-p56lck, anti-LAT and anti-CD3-ζ antibodies.

Article Snippet: The Src family PTK inhibitor PP2, PMA and ionomycin were from Calbiochem (La Jolla, CA).

Techniques: Activity Assay, Immunoprecipitation, Incubation