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Purified anti-SLP76 Phospho (Tyr128) Antibody

Pricing & Availability
Clone
A20007B (See other available formats)
Regulatory Status
RUO
Other Names
Lymphocyte cytosolic protein 2 (LCP2), SH2 domain-containing leukocyte protein of 76 kDa (SLP76), IMD81
Isotype
Mouse IgG1, κ
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Product Citations
publications
A. A20007B_PURE_LP76_ICC_061223
Jurkat cells left untreated (panels A and B) or treated with H2O2 (panels C and D) were fixed with 4% PFA Fixation Buffer (Cat. No. 420801) for 10 minutes and permeabilized with 100% ice-cold methanol for 10 minutes, and blocked with 5% FBS for 1 hour at room temperature. Cells were then stained with 2.5 µg/mL of purified anti-SLP76 Phospho (Tyr128) (clone A20007B), followed by incubation with Alexa Fluor® 594 goat anti-mouse IgG (Cat. No. 405326) for 1 hour at room temperature. Nuclei were counterstained with DAPI (panels A and C)(Cat. No. 422801). The images were captured in a Revvity Operetta CLS™ High Content Analysis System with a 40X objective. Scale bar: 25 µm
  • A. A20007B_PURE_LP76_ICC_061223
    Jurkat cells left untreated (panels A and B) or treated with H2O2 (panels C and D) were fixed with 4% PFA Fixation Buffer (Cat. No. 420801) for 10 minutes and permeabilized with 100% ice-cold methanol for 10 minutes, and blocked with 5% FBS for 1 hour at room temperature. Cells were then stained with 2.5 µg/mL of purified anti-SLP76 Phospho (Tyr128) (clone A20007B), followed by incubation with Alexa Fluor® 594 goat anti-mouse IgG (Cat. No. 405326) for 1 hour at room temperature. Nuclei were counterstained with DAPI (panels A and C)(Cat. No. 422801). The images were captured in a Revvity Operetta CLS™ High Content Analysis System with a 40X objective. Scale bar: 25 µm
  • B.A20007B_PURE_LP76_ICFC_061223-
    Jurkat cells untreated (low expression control, open histogram), or treated with H2O2 for 5 minutes (positive control, filled histogram) were fixed and permeabilized using the Cyto-Fast™ Fix/Perm Buffer Set (Cat. No. 426803) and intracellularly stained with purified anti-SLP76 Phospho (Tyr128) (clone A20007B) or mouse IgG1, κ isotype control (open histogram, dashed line) (representative histogram for untreated or treated cells) (Cat. No. 401401) followed by PE goat anti-mouse IgG (Cat. No. 405307).
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618851 25 µg £133
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618852 100 µg £264
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Description

SLP76 is a cytosolic adaptor protein that functions as a substrate in the T cell receptor (TCR) activation pathway. Upon TCR engagement, SLP76 is phosphorylated on multiple residues by ZAP70, resulting in the protein being translocated to the plasma membrane and facilitating the assembly of a multi-subunit complex comprised of Vav1, Nck, ITK, and PLCγ1 that activates multiple signaling pathways. Mutation and inactivation of the phosphorylation sites results in abrogation of SLP76 activity. Tyr128 is one of three residues on the N-terminus of SLP76 which becomes phosphorylated upon TCR ligation, which initiates immune cell synapse formation. SLP76 also plays an essential role in normal T cell development, and mast cell and platelet function. 

Product Details
Technical Data Sheet (pdf)

Product Details

Verified Reactivity
Human
Antibody Type
Monoclonal
Host Species
Mouse
Immunogen
Synthetic human SLP76 peptide phosphorylated at Tyr128
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

ICC - Quality tested
ICFC - Verified
Recommended Usage

Each lot of this antibody is quality control tested by immunocytochemistry. For immunocytochemistry, a concentration range of 2.5 - 5.0 μg/mL is recommended. For flow cytometric staining, the suggested use of this reagent is ≤ 0.125 µg per million cells in 100 µL volume. It is recommended that the reagent be titrated for optimal performance for each application.

Application Notes

Clone is not suitable for Western Blot application. Although the antibody does detect phospho-SLP76, there is a prominent non-specific band around 35 kDa.

For immunocytochemistry, successful staining was accomplished using fixation with 4% PFA for 10 minutes followed by permeabilization with either 100% ice-cold methanol or 0.5% Triton-X for 10 minutes, or with fixation/permeabilization with 100% ice-cold methanol for 10 minutes. 

For flow cytometry, we recommend treatment with Cyto-Fast™ Fix/Perm Buffer Set (Cat. No. 426803).

RRID
AB_3083403 (BioLegend Cat. No. 618851)
AB_3083403 (BioLegend Cat. No. 618852)

Antigen Details

Structure
SLP76 is a 533 amino acid protein with a predicted molecular weight of ~60 kD.
Distribution

Hematopoietic cells/cytosol

Function
Adaptor protein/TCR Signaling
Interaction
ZAP70, ITK, LAT, GRB2, PLCγ1, NCK, VAV, ITK, LCK, 14-3-3ε
Cell Type
B cells, Monocytes, NK cells, T cells
Biology Area
Cell Biology, Immunology, Signal Transduction
Molecular Family
Phospho-Proteins, Protein Kinases/Phosphatase
Antigen References
  1. Sauer K, et al. 2001. J Biol Chem. 276:45207-16.
  2. Koretzky GA, et al. 2006. Nat Rev Immunol. 6:67-78. 
  3. Di Bartolo V, et al. 2007. J Exp Med. 204:681-91.
  4. Lasserre, et al. 2011. J Cell Biol. 195:839-53.
  5. Wardenberg, et al. 1996. J Biol Chem. 271:33.
Gene ID
3937 View all products for this Gene ID
UniProt
View information about SLP76 Phospho (Tyr128) on UniProt.org

Related FAQs

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Go To Top Version: 1    Revision Date: 06/12/2023

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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