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Purified anti-LAT Phospho (Tyr171) Antibody

Pricing & Availability
Clone
A20005D (See other available formats)
Regulatory Status
RUO
Other Names
Linker For Activation Of T Cells; P36-38
Isotype
Mouse IgG2b, κ
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Product Citations
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A20005D_PURE_LAT_Antibody_1_111621
Whole cell extracts (15 μg protein) from serum-starved Jurkat and EL4 cells untreated (-) or treated (+) with 5 mM H2O2 for 3 minutes, were resolved on a 4-12% Bis-Tris gel, transferred to a PVDF membrane, and probed with 0.0125 μg/mL (1:1000 dilution) of purified anti-LAT Phospho (Tyr171) (clone A20005D) for 2 hours at room temperature. Proteins were visualized by chemiluminescence detection using HRP goat anti-mouse IgG antibody (Cat. No. 405306) at a 1:3000 dilution. Equal LAT loading was confirmed by probing membranes with anti-LAT antibody at a 1:1000 dilution. Western-Ready™ ECL Substrate Premium Kit (Cat. No. 426319) was used as a detection agent. Direct-Blot™ HRP anti-β-actin antibody (Cat. No. 643807) was used as a loading control at a 1:10000 dilution (lower). Lane M: Molecular weight marker
  • A20005D_PURE_LAT_Antibody_1_111621
    Whole cell extracts (15 μg protein) from serum-starved Jurkat and EL4 cells untreated (-) or treated (+) with 5 mM H2O2 for 3 minutes, were resolved on a 4-12% Bis-Tris gel, transferred to a PVDF membrane, and probed with 0.0125 μg/mL (1:1000 dilution) of purified anti-LAT Phospho (Tyr171) (clone A20005D) for 2 hours at room temperature. Proteins were visualized by chemiluminescence detection using HRP goat anti-mouse IgG antibody (Cat. No. 405306) at a 1:3000 dilution. Equal LAT loading was confirmed by probing membranes with anti-LAT antibody at a 1:1000 dilution. Western-Ready™ ECL Substrate Premium Kit (Cat. No. 426319) was used as a detection agent. Direct-Blot™ HRP anti-β-actin antibody (Cat. No. 643807) was used as a loading control at a 1:10000 dilution (lower). Lane M: Molecular weight marker
  • A20005D_PURE_LAT_Antibody_2_111621
    Serum-starved Jurkat cells untreated (lower) and treated (upper) with 5 mM H2O2 for 3 minutes, were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with Triton X-100 for 10 minutes, and blocked with 5% FBS for 60 minutes. Cells were then intracellularly stained with 5.0 µg/mL of either purified mouse IgG2b, κ isotype control (Cat. No. 400302) (panel A and C) or purified anti-LAT Phospho (Tyr171) (clone A20005D) (panel B and D) (1:10 dilution) for 2 hours at room temperature, followed by incubation with Alexa Fluor® 594 goat anti-mouse IgG (Cat. No. 405326) at a 1:200 dilution. Nuclei were counterstained with DAPI and the image was captured with a 60X objective.
  • A20005D_PURE_LAT_Antibody_3_111621
    Jurkat cells treated (filled histogram, positive control) with 5mM of H2O2 for 3 min or untreated (open histogram, negative control) were fixed and permeabilized using the True-Phos™ Perm Buffer set (Cat. No. 425401), and intracellularly stained with 0.125 µg per test (1:20 dilution) purified anti-LAT Phospho (Tyr171) (clone A20005D) followed by PE goat anti-mouse IgG antibody (Cat. No. 405307).
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946601 25 µL 128€
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946602 100 µL 296€
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Description

LAT, or Linker for Activation of T cells, is a transmembrane protein expressed mainly in T cells and a limited number of other immune cells such as natural killer cells, mast cells, and immature B cells. As an adaptor protein, phosphorylated LAT facilitates the recruitment of other signaling proteins and functions as a nucleating site for multiprotein signaling complexes. These signaling complexes can propagate TCR, or T Cell antigen Receptors, and activate downstream effectors, which trigger T cell proliferation or cytokine expression.

LAT (Tyr171) is one of the nine tyrosines in LAT conserved between human, mice, and rat. LAT (Tyr171) is necessary for the activation of PI 3-kinase, a protein that regulates the recruitment of other proteins that facilitate in T-cell receptor signal transduction such as PLC-gamma1 and Sos1. When LAT (Tyr171) and (Tyr191) is phosphorylated by ZAP-70, Gads can bind directly to LAT (Tyr171) and (Tyr191) and mediate the recruitment of SLP-76. 

Product Details
Technical Data Sheet (pdf)

Product Details

Verified Reactivity
Human, Mouse
Antibody Type
Monoclonal
Host Species
Mouse
Immunogen
Synthetic peptide corresponding to human LAT phosphorylated at Tyr 171
Formulation
Phosphate Buffer, 0.5% BSA, 0.09% NaN3
Preparation
The antibody was purified by affinity chromatography.
Concentration
0.05 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 dilution of this reagent is 1000 - 1:5000. For immunocytochemistry, a dilution of 1:10 is recommended. For intracellular flow cytometric staining, the suggested dilution of this reagent is 1:20 per million cells in 100 µL volume. It is recommended that the reagent be titrated for optimal performance for each application.

Application Notes

When this clone is used in WB at a dilution lower than 1:1000 it may show non-specific bands at 50 kD and above.

This clone has been tested by ICC on H2O2 treated and untreated Jurkat, with three fix and permeabilization methods (100% methanol, 4% PFA plus methanol and 4% PFA plus Triton X-100). All three methods can be used for ICC staining, but the PFA plus Triton X-100 fixation/permeabilization method produced the best staining.

This clone has only been tested on True-PhosTM perm buffer (Cat. No. 425401) by ICFC

RRID
AB_2904456 (BioLegend Cat. No. 946601)
AB_2904456 (BioLegend Cat. No. 946602)

Antigen Details

Structure
LAT is a 262 amino acid protein with a predicted molecular weight of 36-38 kD.
Distribution

Plasma membrane/T Cells

Function
Plasma membrane/T Cells
Cell Type
Mast cells, NK cells, T cells
Biology Area
Cell Biology, Immunology, Signal Transduction
Molecular Family
Phospho-Proteins
Antigen References
  1. Balagopalan L. et al., 2010, Cold Spring Harb Perspect Biol., 2(8): a005512
  2. Balagopalan L. et al., 2020, PLoS One, 15(2): e0229036
  3. Lin J. and Weiss A., 2001, J Biol Chem., 276(31): 29588-95
  4. Paz et. al., 2001, Biochem J., 356, 461-71
  5. Zhang W. et al., 2000, J Biol Chem., 275(30): 23355-61
Gene ID
27040 View all products for this Gene ID
UniProt
View information about LAT Phospho Tyr171 on UniProt.org

Related FAQs

There are no FAQs for this product.
Go To Top Version: 2    Revision Date: 06.15.2022

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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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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