Thermo Fisher Scientific Phospho-LCK (Tyr505) Monoclonal Antibody (SRRCHA), eFluor 660, eBioscience

Thermo Fisher Scientific Phospho-LCK (Tyr505) Monoclonal Antibody (SRRCHA), eFluor 660, eBioscience

Applications

• Flow Cytometry (Intracellular staining)

Tested Dilution: 5 µL (0.125 µg)/test

Product Specifications

Product Specific Information

This SRRCHA monoclonal antibody recognizes human and mouse LCK when phosphorylated on tyrosine 505 (Y505). Y505 is a negative-regulatory site phosphorylated by CSK. LCK is expressed in T cells, associating with CD4 and CD8 co-receptors. Upon TCR engagement, LCK phosphorylates CD3 and zeta chains, enabling ZAP-70 binding and activation. Activated ZAP-70 phosphorylates LAT, recruiting signaling proteins including SLP-76, Grb2, and PLCγ1. LCK deficiency leads to impaired T cell development and function.

Applications Reported: Intracellular staining followed by flow cytometric analysis.
Applications Tested: Pre-titrated for intracellular staining of stimulated human peripheral blood cells. Recommended use: 5 µL (0.125 µg) per 100 µL test volume.

Fluorochrome Information:
eFluor® 660 replaces Alexa Fluor® 647. Excitation: 633–647 nm, Emission: 668 nm, compatible with red laser detection.

Staining Protocols:

• Protocol A: Two-step for cytoplasmic proteins (surface + intracellular detection)
• Protocol B: One-step for nuclear proteins (transcription factor staining)
• Protocol C: Two-step fixation/methanol for phospho-specific signaling (limited surface staining)

Refer to BestProtocols®: Staining Intracellular Antigens for Flow Cytometry.

Filtration: 0.2 µm post-manufacturing filtered.

Target Information

LCK is a Src-family tyrosine kinase essential for T-cell receptor (TCR) signaling. Upon TCR activation, LCK phosphorylates ITAM motifs, enabling ZAP70 binding and activation, which leads to LAT phosphorylation and downstream signaling. LCK localizes mainly at the plasma membrane and partially at the Golgi apparatus. It contains SH2/SH3 domains for protein-protein interactions and regulates apoptosis in non-death receptor pathways. Associated diseases include Immunodeficiency 22 and CD45 deficiency. Several splice variants of LCK exist.

For Research Use Only. Not for use in diagnostic procedures or resale without authorization.

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