Western blot analysis of extracts from WEHI-231 cells, untreated (-) or treated (+) with combinations of the following treatments as indicated: H2O2 (100 mM, 20 min), Ibrutinib pretreatment (1 μM, 1 hr), using Phospho-Btk (Tyr223) (D9T6H) Rabbit mAb #87141, Btk (D3H5) Rabbit mAb #8547 and β-Actin (D6A8) Rabbit mAb #8457.
Chemical structure of Ibrutinib.
Ibrutinib is supplied as a lyophilized powder. For a 10mM stock, reconstitute the 5 mg of powder in 1.14 ml DMSO. Working concentrations and length of treatment can vary depending on the desired effect, but it is typically used as a pretreatment at 1-10 μM for 1 hour prior to treating with a stimulator.
Store lyophilized or in solution at -20C, dessicated. Protect from light. In lyophilized form, the chemical is stable for 24 months. Once in solution, use within 3 months to prevent loss of potency. Aliquot to avoid mulitple freeze/thaw cycles.
Ibrutinib, also known as PCI-32765, is a small molecule inhibitor of Bruton's tyrosine kinase (Btk) (1,2). Research studies demonstrate that ibrutinib inhibits autophosphorylation of Btk as well as downstream targets such as PLCγ2 and ERK by binding to Cys481 in the active site (1,2). The inhibition of Btk also prevents NFκB DNA binding, reduces cell migration, proliferation and survival, inhibits DNA synthesis and induces apopotosis (1).
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