|Detection of Human Lck by Western Blot. Western blot shows lysates of Jurkat human acute T cell leukemia cell line and COLO 205 human colorectal adenocarcinoma cell line. PVDF membrane was probed with 0.2 µg/mL of Mouse Anti-Human Lck Monoclonal Antibody (Catalog # MAB37041) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for Lck at approximately 56 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.|
Intracellular Staining by Flow Cytometry
|Detection of Lck in Human Blood Lymphocytes by Flow Cytometry. Human peripheral blood lymphocytes were stained with Mouse Anti-Human Lck Monoclonal Antibody (Catalog # MAB37041) followed by Phycoerythrin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0102B) and Mouse Anti-Human CD3 epsilon APC‑conjugated Monoclonal Antibody (Catalog # FAB100A). Quadrant markers were set based on control antibody staining (Catalog # MAB003).|
|Lck in Human PBMCs. Lck was detected in immersion fixed human peripheral blood mononuclear cells (PBMCs) using Mouse Anti-Human Lck Monoclonal Antibody (Catalog # MAB37041) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm and cell surfaces. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.|
Lck (p56lck) is a 56 kDa cytosolic phosphoprotein within the Src family of non-receptor tyrosine kinases. The 509 amino acid (aa) human Lck contains an SH3 domain (aa 61-121), an SH2 domain (aa 127-224) and a protein kinase domain (aa 251-259). Within aa 2-66, human Lck shares 89% aa sequence identity with mouse and rat Lck. A short (363 aa) isoform contains alternate sequence starting at aa 348, while a 539 aa isoform contains inserted sequence after aa 321. Lck interacts with T cell CD4 and CD8 molecules and plays a pivotal role in regulating T cell activation. It can be overexpressed in cancers and functions as an oncogene.