Mouse Monoclonal LCK Antibody

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     Anti-LCK Antibody at 1:2000 dilution + Jurkat whole cell lysate Lysates/proteins at 20 μg per lane. Secondary Goat Anti-mouse IgG,  (H+L), Peroxidase conjugated at 1/10000 dilution. Predicted band size : 58 kDa Blocking/Dilution buffer: 5% NFDM/TBST.    

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     Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human cervical epithelial adenocarcinoma cell line) cells labeling LCK with P33312 at 1/25 dilution, followed by Dylight® 488-conjugated goat anti-mouse IgG   secondary antibody at 1/200 dilution (green). Immunofluorescence image showing cytoplasm staining on HeLa cell line. Cytoplasmic actin is detected with Dylight® 554 Phalloidin  at 1/100 dilution (red).The nuclear counter stain is DAPI (blue).    

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     Overlay histogram showing Jurkat cells stained with P33312 (green line).  The cells were fixed with 2% paraformaldehyde (10 min) and then permeabilized with 90% methanol for 10 min.  The cells were then icubated in 2% bovine serum albumin to block non-specific protein-protein interactions followed by the antibody (P33312,  1:25 dilution) for 60 min at 37ºC.  The secondary antibody used was Goat-Anti-Mouse IgG, DyLight® 488 Conjugated Highly Cross-Adsorbed(NA168821) at 1/400 dilution for 40 min at 37ºC.  Isotype control antibody (blue line) was mouse IgG (1μg/1x10^6 cells) used under the same conditions.  Acquisition of >10, 000 events was performed.    

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     P33312 staining LCK in human thymus sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed,  paraffin-embedded sections).  Tissue was fixed with formaldehyde and blocked with 3% BSA for 0. 5 hour at room temperature; antigen retrieval was by heat mediation with a citrate buffer (pH6).  Samples were incubated with primary antibody (1/25) for 1 hours at 37°C.  A undiluted biotinylated goat polyvalent antibody was used as the secondary antibody.    

LCK (lymphocyte-specific protein tyrosine kinase) is a member of the Src family of tyrosine kinases, primarily expressed in T cells and natural killer (NK) cells. It plays a critical role in T-cell receptor (TCR) signaling, facilitating early activation events by phosphorylating downstream targets like ZAP-70 and adaptor proteins. LCK is anchored to the plasma membrane via its N-terminal lipid modification and interacts with TCR-CD3 complexes through co-receptors like CD4 or CD8. This proximity allows LCK to initiate signaling cascades upon antigen recognition, regulating T-cell development, activation, and differentiation.

Antibodies targeting LCK are essential tools in immunology research. They are used to study protein expression, localization, and function in T-cell signaling pathways. Applications include Western blotting, flow cytometry, and immunohistochemistry to explore LCK's role in immune responses, autoimmune diseases, and cancers. Dysregulation of LCK activity has been implicated in T-cell malignancies and immunodeficiency disorders, making it a potential therapeutic target. Researchers also use LCK antibodies to investigate mechanisms of immunosuppressive drugs or novel immunotherapies. Commercial LCK antibodies are typically validated for specificity using knockout cell lines or functional assays, with monoclonal and polyclonal variants available for different experimental needs.