Antibody data

Product number
50-9076-42 - Provider product page
Provider
Invitrogen Antibodies
Product name
Phospho-LCK (Tyr505) Monoclonal Antibody (SRRCHA), eFluor™ 660, eBioscience™
Antibody type
Monoclonal
Antigen
Synthetic peptide
Description
Description: This SRRCHA monoclonal antibody recognizes human and mouse LCK when phosphorylated on tyrosine 505 (Y505). Y505 is a negative-regulatory site that is phosphorylated by the kinase CSK. LCK is expressed in T cells where it associates with the cytoplasmic tails of the CD4 and CD8 co-receptors on T helper cells and cytotoxic T cells, respectively. When the TCR is engaged, LCK acts to phosphorylate the intracellular chains of CD3 and zeta chains of the TCR complex, allowing the cytoplasmic tyrosine kinase, ZAP-70, to bind to them. LCK then phosphorylates and activates ZAP-70, which then phosphorylates the adaptor protein, LAT, that serves as a docking site for a number of other proteins, including SLP-76, Grb2, and phospholipase C gamma1. Mice lacking expression of LCK show defects in T cell development and function. Applications Reported:This SRRCHA antibody has been reported for use in intracellular staining followed by flow cytometric analysis. Applications Tested: This SRRCHA antibody has been pre-titrated and tested by intracellular staining followed by flow cytometric analysis of stimulated normal human peripheral blood cells. This can be used at 5 µL (0.125 µg) per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. eFluor® 660 is a replacement for Alexa Fluor® 647. eFluor® 660 emits at 659 nm and is excited with the red laser (633 nm). Please make sure that your instrument is capable of detecting this fluorochrome. Staining Protocol: All protocols work well for this monoclonal antibody. Use of Protocol A: Two-step protocol: intracellular (cytoplasmic) proteins allows for the greatest flexibility for detection of surface and intracellular (cytoplasmic) proteins. Use of Protocol B: One-step protocol: intracellular (nuclear) proteins is recommended for staining of transcription factors in conjunction with surface and phosphorylated intracellular (cytoplasmic) proteins. Protocol C: Two-step protocol: Fixation/Methanol allows for the greatest discrimination of phospho-specific signaling between unstimulated and stimulated samples, but with limitations on the ability to stain specific surface proteins (refer to "Clone Performance Following Fixation/Permeabilization" located in the BestProtocols Section under the Resources tab online). All Protocols can be found in the Flow Cytometry Protocols: "Staining Intracellular Antigens for Flow Cytometry Protocol" located in the BestProtocols® Section under the Resources tab online. Excitation: 633-647 nm; Emission: 668 nm; Laser: Red Laser. Filtration: 0.2 µm post-manufacturing filtered.
Reactivity
Human, Mouse
Host
Mouse
Isotype
IgG
Antibody clone number
SRRCHA
Vial size
100 Tests
Concentration
5 μL/Test
Storage
4°C, store in dark, DO NOT FREEZE!
LCK protein structure - 50-9076-42 shown in red.