Detection of Human BLNK by Western Blot.
Western blot shows lysates of Daudi human Burkitt's lymphoma cell line, Raji human Burkitt's lymphoma cell line, Ramos human Burkitt's lymphoma cell line, BJAB human Burkitt's lymphoma cell line, and Jurkat human acute T cell leukemia cell line. PVDF membrane was probed with 1 µg/mL of Human BLNK Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4966) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). A specific band was detected for BLNK at approximately 70 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
BLNK in Human Spleen.
BLNK was detected in immersion fixed paraffin-embedded sections of human spleen using Goat Anti-Human BLNK Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4966) at 10 µg/mL overnight at 4 °C. Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to splenocytes. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Detection of Human BLNK by Simple WesternTM.
Simple Western lane view shows lysates of Ramos human Burkitt's lymphoma cell line and Raji human Burkitt's lymphoma cell line, loaded at 0.2 mg/mL. A specific band was detected for BLNK at approximately 81 kDa (as indicated) using 10 µg/mL of Goat Anti-Human BLNK Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4966) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
Reconstitution Buffer Available
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
BLNK (B cell linker protein; also SLP-65 and BASH) is a 50 kDa (predicted) cytoplasmic protein that runs anomalously at 65‑70 kDa in SDS‑PAGE. It is a B cell adapter molecule that, following phosphorylation by Syk, serves as a scaffold for assembly of macromolecular complexes that involve Grb2, Vav and PLC gamma. Tyrosine phosphorylation occurs at Tyr72/84/96/178/189. Human BLNK is 456 amino acids (aa) in length and it contains a Pro-rich region (aa 98‑260) and one SH2 domain (aa 346‑453). More than 10 potential splice variants have been detected and among these are four which show single in-frame deletions between aa 203‑225, 366‑417, 146‑417 and 211‑456, respectively. Over aa 226‑456, human BLNK shares 87% and 83% aa identity with canine and mouse BLNK, respectively.
B-cell Linker Protein
Entrez Gene IDs:
29760 (Human); 17060 (Mouse); 499356 (Rat)
AGM4; B cell linker protein; BASH; B-cell adapter containing a SH2 domain protein; B-cell adapter containing a Src homology 2 domain protein; B-cell linker protein; B-cell linker; BLNK; BLNK-s; Cytoplasmic adapter protein; Ly57; MGC111051; SLP65; SLP-65BLNK-S; SLP65Ly57; Src homology 2 domain-containing leukocyte protein of 65 kDa
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