Human BLNK Antibody

Catalog # Availability Size / Price Qty
AF4966
AF4966-SP

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Detection of Human BLNK by Western Blot.
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Human BLNK Antibody Summary

Species Reactivity
Human
Specificity
Detects endogenous human BLNK in Western blots.
Source
Polyclonal Goat IgG
Purification
Antigen Affinity-purified
Immunogen
E. coli-derived recombinant human BLNK
Gly226-Ser456
Accession # Q8WV28
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
1 µg/mL
See below
Simple Western
10 µg/mL
See below
Immunohistochemistry
5-15 µg/mL
See below

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Western Blot Detection of Human BLNK antibody by Western Blot. View Larger

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.

Immunohistochemistry BLNK antibody in Human Spleen by Immunohistochemistry (IHC-P). View Larger

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.

Simple Western Detection of Human BLNK antibody by Simple Western<sup>TM</sup>. View Larger

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.

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
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Shipping
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.

Background: BLNK

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.

Long Name
B-cell Linker Protein
Entrez Gene IDs
29760 (Human); 17060 (Mouse); 499356 (Rat)
Alternate Names
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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