Antibodies to B-cell Linker Protein (BLNK)

Introduction

Linker or adapter proteins provide mechanisms by which receptors can amplify and regulate downstream effector proteins. BLNK (also referred to as BASH or SLP-65) is an adaptor molecule that plays a key role in B cell activation initiated by cross-linking the B cell receptor (BCR).

This activation leads to an increase in the tyrosine phosphorylation of a number of proteins due to activation of non-receptor protein tyrosine kinases (PTK) including Syk, which is responsible for the tyrosine phosphorylation of BLNK. BLNK is a cytosolic protein identified as two phosphoproteins migrating at 68 and 70 kDa in SDS/PAGE (alternatively spliced forms of human BLNK). Mapping of the tyrosine residues in BLNK reveal three tyrosines that bind PLC-gamma2, and two tyrosines that bind the Tec family kinase Btk, the Vav nucleotide exchange factor and the adaptor Nck. BLNK regulates B cell proliferation and differentiation; the recently reported complete loss or drastic reduction of BLNK expression in ~50% of human childhood pre-B acute lympho¬blastic leukaemias (ALL) may be one example for its crucial function.

Antibody Panel to B-cell Linker Protein (BLNK)

Acris Antibodies offers a panel of monoclonal and polyclonal antibodies for the detection of BLNK in human, rat or mouse tissues. Antibodies can be used for Immunohistochemistry, Western blotting or other Immunological methods.

Fig.2: Peptide competition experiment: EB1 cells were serum starved and left unstimulated (1) or stimulated (2-5) with 3 mM H2O2, and cell lysates were resolved by SDS-PAGE on a 10% Tris-glycine gel and transferred to PVDF. Membranes were blocked with a 5% BSA-TBST buffer overnight at 4oC, then were incubated with 0.50 µg/mL BLNK [pTyr84] antibody for two hours at room temperature in a 3% BSA-TBST buffer, following prior incubation with: no peptide (1, 2), the non-phosphopeptide corresponding to the immunogen (3), a generic phosphotyrosine containing peptide (4), or, the phosphopeptide immunogen (5). After washing, membranes were incubated with goat F(ab’)2 anti-rabbit IgG alkaline phosphatase and signals were detected using the Tropix WesternStar‘ method. The data show that only the peptide corresponding to BLNK [pTyr84] blocks the antibody signal, thereby demonstrating the specificity of the antibody, and H2O2 induced tyrosine phosphorylation of BLNK [pTyr84].

Antibody Panel to BLNK

B-cell Linker Protein (BLNK), BASH, SLP-65, antibody.

Literature

Xu S, Huo J, Chew WK, Hikida M, Kurosaki T, Lam KP. Phospholipase Cgamma2 dosage is critical for B cell development in the absence of adaptor protein BLNK. J Immunol. 2006 Apr 15;176(8):4690-8.

Gupta N, Delrow J, Drawid A, Sengupta AM, Fan G, Gélinas C. Repression of B-cell linker (BLNK) and B-cell adaptor for phosphoinositide 3-kinase (BCAP) is important for lymphocyte transformation by rel proteins. Cancer Res. 2008 Feb 1;68(3):808-14.

Chui, C.W., et al. (2002) BLNK: molecular scaffolding through ‘cis’-mediated organization of signaling proteins. EMBO J. 23:6461-6472 (cites the use of this antibody in Western blotting).

Kabak, S., et al. (2002) The direct recruitment of BLNK to immunoglobulin alpha couples the B-cell antigen receptor to distal signaling pathways. Mol. Cell. Biol. 22(8):2524-2535.

Taguchi, T. et al. (2004) Deficiency of BLNK hampers PLC-gamma 2 phosphorylation and Ca2+ influx induced by the pre-B cell receptor in human pre-B cells.Immunology 112(4): 575-582.

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