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Purified anti-BLNK Phospho (Tyr96) Antibody

Pricing & Availability
Clone
A19010A (See other available formats)
Regulatory Status
RUO
Other Names
B Cell Linker (BLNK), B-Cell Adapter Containing ASH2 Domain Protein, SLP65, BASH
Isotype
Mouse IgG1, κ
Ave. Rating
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Product Citations
publications
A_A19010A_PURE_BLNK_Antibody_WB_112222
Whole cell extracts (15 µg total protein) from serum-starved Ramos cells untreated (-) or treated (+) with anti-IgM (12 µg/mL,15 min) and H2O2 (5 mM, 3 min), were resolved on a 4-12% Bis-Tris gel, transferred to a PVDF membrane, and probed with 0.1 µg/mL of purified anti-BLNK Phospho (Tyr96) (clone A19010A) for 2 hours at 4°C. Proteins were visualized by chemiluminescence detection using HRP goat anti-mouse IgG (Cat. No. 405306). Equal BLNK loading was confirmed by probing membranes with anti-pan BLNK antibody. Direct-Blot™ HRP anti-β-Actin (Cat. No. 643807) was used as a loading control. Western-Ready™ ECL Substrate Premium Kit (Cat. No. 426319) was used as a detection reagent. Lane M: Molecular weight marker
  • A_A19010A_PURE_BLNK_Antibody_WB_112222
    Whole cell extracts (15 µg total protein) from serum-starved Ramos cells untreated (-) or treated (+) with anti-IgM (12 µg/mL,15 min) and H2O2 (5 mM, 3 min), were resolved on a 4-12% Bis-Tris gel, transferred to a PVDF membrane, and probed with 0.1 µg/mL of purified anti-BLNK Phospho (Tyr96) (clone A19010A) for 2 hours at 4°C. Proteins were visualized by chemiluminescence detection using HRP goat anti-mouse IgG (Cat. No. 405306). Equal BLNK loading was confirmed by probing membranes with anti-pan BLNK antibody. Direct-Blot™ HRP anti-β-Actin (Cat. No. 643807) was used as a loading control. Western-Ready™ ECL Substrate Premium Kit (Cat. No. 426319) was used as a detection reagent. Lane M: Molecular weight marker
  • B_A19010A_PURE_BLNK_Antibody_ICC_112222
    Serum-starved Ramos cells were treated with (right panels) or without (left panels) anti-IgM (12 µg/mL, 15 min) and H2O2 (5 mM, 3 min) fixed with 4% paraformaldehyde for 10 minutes, permeabilized with methanol for 10 minutes, and blocked with 5% FBS for 60 minutes. The cells were then overnight incubated with 5 µg/mL of purified anti-BLNK Phospho (Tyr96) (clone A19010A) (bottom panel) or purified mouse IgG1, κ isotype control (Cat. No. 400102) (top panel) at 4°C, followed by incubation with 2 µg/mL of Alexa Fluor® 594 goat anti-mouse IgG (Cat. No. 405326) for one hour at room temperature. Nuclei were counterstained with DAPI, and the slides were mounted with ProLong™ Gold Antifade Mountant. The image was captured with a 60X objective. Scale bar: 50 µm
  • C_A19010A_PURE_BLNK_Antibody_ICFC_120722
    Human peripheral blood lymphocytes were treated with (left panel), or without (right panel) anti-IgM (12 µg/mL, 15 min) and H2O2 (5 mM, 3 min), fixed with Fixation Buffer (Cat. No. 420801), permeabilized with True-Phos™ Perm Buffer (Cat. No. 425401), then stained intracellularly with purified anti-BLNK Phospho (Tyr96) (clone A19010A) followed by anti-mouse IgG PE. Cells were co-stained with anti-human CD20 APC.
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608351 25 µg 134 CHF
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608352 100 µg 400 CHF
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Description

B cell Linker protein (BLNK) is a cytoplasmic adaptor protein that is involved in B cell antigen receptor (BCR) signaling, a mechanism that is essential for the development of B lymphocytes in the bone marrow and periphery. Following BCR activation, BLNK undergoes tyrosine phosphorylation by Syk which induces interaction with effector molecules such as Grb2, Vav, and PLCγ to facilitate downstream signaling pathways. Phosphorylation sites include Tyr72, Tyr84, Tyr96, Tyr178, Tyr189, and Tyr394. Phosphorylation at Tyr96 residue facilitates BTK binding and plays a critical role in B cell maturation. BLNK mutations can block pro- to pre-B cell transitioning and result in gammaglobulinemia and absent B cells.

Product Details
Technical Data Sheet (pdf)

Product Details

Verified Reactivity
Human
Antibody Type
Monoclonal
Host Species
Mouse
Immunogen
Synthetic peptide corresponding to human BLNK phosphorylated at Tyrosine 96
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide
Preparation
The antibody was purified by affinity chromatography.
Concentration
0.5 mg/mL
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C.
Application

WB - Quality tested
ICC, ICFC - Verified
Recommended Usage

Each lot of this antibody is quality control tested by western blotting. For western blotting, the suggested use of this reagent is 0.1 µg/mL. For immunocytochemistry, a concentration range of 1.25 - 5.0 μg/mL is recommended. For flow cytometric staining, the suggested use of this reagent is ≤ 0.25 µg per million cells in 100 µL volume. It is recommended that the reagent be titrated for optimal performance for each application.

Application Notes

This product has high background in WB if used at concentrations above 0.2 μg/mL.

This product is not suitable for ICC if cells are fixed with methanol only.

RRID
AB_2936688 (BioLegend Cat. No. 608351)
AB_2936688 (BioLegend Cat. No. 608352)

Antigen Details

Structure
BLNK is a 456 amino-acid protein with a predicted molecular weight of 50 kD.
Distribution

BLNK is expressed in cytoplasm of B cells.

Function
Activation of BLNK plays a critical role in B cell proliferation, differentiation, and apoptosis.
Interaction
BLNK interacts with Grb2, Vav, PLCγ2, and Nck1.
Cell Type
B cells
Biology Area
Cell Biology
Molecular Family
Adaptor Proteins, Phospho-Proteins
Antigen References
  1. Fu C, et al. 1998. Immunity. 9:93-103.
  2. Kurosaki T & Tsukada S. 2000. Immunity. 12:1-5.
  3. Gauld SB, et al. 2002. Science. 296:1641-2.
  4. Minegishi Y, et al. 1999. Science. 286:1954-7.
Gene ID
29760 View all products for this Gene ID
UniProt
View information about BLNK Phospho Tyr96 on UniProt.org
Go To Top Version: 2    Revision Date: 04.21.2023

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

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This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
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