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Western blot analysis of BLNK (pY84) in human Burkitt's lymphoma. Lysates from control (lanes 1-3) and hydrogen peroxide-activated (lanes 4-6) Ramos cells were probed with purified mouse anti-BLNK (pY84) monoclonal antibody) at concentrations of 0.125 (lanes 1 and 4), 0.0625 (lanes 2 and 5), and 0.032 µg/ml (lanes 3 and 6). BLNK (pY84) is identified as a band of about 68 kDa in the treated cells.
BD Pharmingen™ Purified Mouse anti-BLNK (pY84)
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- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
- Sodium azide is a reversible inhibitor of oxidative metabolism; therefore, antibody preparations containing this preservative agent must not be used in cell cultures nor injected into animals. Sodium azide may be removed by washing stained cells or plate-bound antibody or dialyzing soluble antibody in sodium azide-free buffer. Since endotoxin may also affect the results of functional studies, we recommend the NA/LE (No Azide/Low Endotoxin) antibody format, if available, for in vitro and in vivo use.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
B cell activation is initiated by crosslinking the B cell receptor, which leads to activation of non-receptor protein tyrosine kinases (PTK), including Btk, Syk, and three Src kinases, Fyn, Lyn, and Blk. Activated PTKs then phosphorylate multiple cellular proteins involved in B lymphocyte signaling. Syk is responsible for the tyrosine phosphorylation of B cell linker protein (BLNK), a member of the SLP-76 family of adapter proteins. Phosphorylation of human BLNK at tyrosines 84, 178, and 189 (Y84, Y178, and Y189) creates docking sites for PLCγ2, leading to the activation of downstream signaling pathways.
The J117-1278 monoclonal antibody recognizes the phosphorylated Y84 of BLNK.
研发参考 (5)
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Chiu CW, Dalton M, Ishiai M, Kurosaki T, Chan AC. BLNK: molecular scaffolding through 'cis'-mediated organization of signaling proteins. EMBO J. 2002; 21:6461-6472. (Biology).
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Espeli M, Mancini S, Breton C, Poirier F, Schiff C . Impaired B-cell development at the pre-Bll-cell stage in galectin-1-deficient mice due to inefficient pre-Bll/stromal cell interactions. Blood. 2009; 113(23):5878-5886. (Clone-specific: Flow cytometry). 查看参考
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Janssen E, Zhang W. Adaptor proteins in lymphocyte activation. Curr Opin Immunol. 2003; 15:269-276. (Biology). 查看参考
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Pighi C, Gu T, Dalai I, et al. Phospho-proteomic analysis of mantle cell lymphoma cells suggests a pro-survival role of B-cell receptor signaling. Cell Oncol. 2011; 34(2):141-153. (Clone-specific: Flow cytometry, Western blot). 查看参考
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Wu JN, Koretzky GA. The SLP-76 family of adapter proteins. Semin Immunol. 2004; 16:379-393. (Biology). 查看参考
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