Invitrogen

LCK Monoclonal Antibody (3A5)

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This Antibody was verified by Relative expression to ensure that the antibody binds to the antigen stated.

1 Reference

View all (44) LCK antibodies

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Datasheet

Protocols

Tech Support

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LCK Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescence analysis of LCK was performed using 70% confluent log phase Jurkat and THP-1 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with LCK Monoclonal Antibody (3A5) (Product # MA5-12303) at 1:100 in 0.1% BSA, incubated at 4-degree Celsius overnight and then labeled with Donkey anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32766), (1:2000), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b: Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing Plasma membrane localization. Panel e represents THP-1 cells which are reported to be low to negative for LCK expression. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

LCK Antibody in Western Blot (WB) — Western blot was performed using Anti-LCK Monoclonal Antibody (3A5) (Product # MA5-12303) and a 55 kDa band corresponding to LCK was observed across was observed in all cells and tissues tested except A549, THP-1, Mouse Brain, Rat Brain and Mouse testis which are reported to be low to negative for LCK expression. An additional uncharacterized band (*) around 110 kDa was observed in Brain tissues, while IgG was observed in Mouse Testis and Mouse Thymus. Membrane enriched extracts (30 µg lysate) of Jurkat (Lane 1), MOLT-4 (Lane 2), A549 (Lane 3), THP-1 (Lane 4), Mouse Brain (Lane 5), Rat Brain (Lane 6), Mouse Testis (Lane 7), Mouse Thymus (Lane 8) and Rat Thymus (Lane 9) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23002) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1ug/ml) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177,1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

LCK Antibody in Flow Cytometry (Flow) — Flow cytometry analysis of LCK (p56lck) in Jurkat cells (green) compared to an isotype control (blue). Cells were harvested, adjusted to a concentration of 1-5x10^6 cells/mL, fixed with 2% paraformaldehyde and washed with PBS. Cells were blocked with a 2% solution of BSA-PBS for 30 min at room temperature and incubated with a LCK (p56lck) monoclonal antibody (Product # MA5-12303) at a dilution of 1 µg/test for 40 min at room temperature. Cells were then incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated secondary antibody and re-suspended in PBS for FACS analysis.

LCK Antibody in Immunoprecipitation (IP) — Immunoprecipitation of LCK (P56lck) using LCK (P56lck) Monoclonal Antibody (Product # MA5-12303) on denatured Human Lck Cells.

LCK Antibody — Lck Protein Schematic-Lck is a 56 kDa, membrane-associated, non-receptor tyrosine kinase belonging to the Src family that plays a critical role in T-cell antigen receptor (TCR) signaling, inducing T-lymphocyte development, activation, proliferation and survival. Lck has been implicated in various diseases including cancer development and metastasis, viral infections relating to HIV, heart disease, and osteoclast-mediated bone resorption. Lck is regulated by phosphorylation on multiple residues including Ser-194 in the SH2 domain, Tyr-394 (autophosphorylation site) within the activation loop of the catalytic domain, and Tyr-505 (inhibitory site) near the C-terminus. TCR requires Lck for thymocyte differentiation and activation. Additionally, Lck is implicated in JAK and STAT signaling, which are constitutively activated in cells overexpressing Lck.

Product Details

MA5-12303

Applications	Tested Dilution	Publications
Western Blot (WB)	1-2 µg/mL	-
Immunocytochemistry (ICC/IF)	1:20-1:100	-
Flow Cytometry (Flow)	1 µg/test	-
Immunoprecipitation (IP)	2 µg/mL	-
Miscellaneous PubMed (Misc)	-	View 1 publication 1 publication

Product Specifications
Species Reactivity	Human, Mouse, Rat
Published species	Not Applicable
Host/Isotype	Mouse / IgG2b, kappa
Class	Monoclonal
Type	Antibody
Clone	3A5
Immunogen	Recombinant protein corresponding to aa 1-225 of murine p56lck protein View immunogen
Conjugate	Unconjugated
Form	Liquid
Concentration	0.2 mg/mL
Purification	Protein A
Storage buffer	PBS, pH 7.4, with 0.2% BSA
Contains	0.09% sodium azide
Storage conditions	4° C
RRID	AB_10986446

Product Specific Information

MA5-12303 targets LCK (p56lck) in IP, ICC/IF, FACS and WB applications and shows reactivity with Human, mouse, and Rat samples.

The MA5-12303 immunogen is recombinant protein corresponding to aa 1-225 of mouse p56lck protein.

Target Information

LCK is a member of the Src-family tyrosine kinase, which are essential for signaling through the T-cell receptor (TCR) complex. Upon TCR triggering, LCK phosphorylates the ITAM motives in its zeta subunits establishing binding sites for the SH2 domains of the tyrosine kinase ZAP70, which is also phosphorylated by LCK and activated to generate subsequent signaling platforms by phosphorylation of adaptor LAT. The majority of LCK is localized to the plasma membrane, however, there is also a significant fraction associated with the Golgi apparatus which may contribute to Raf activation under conditions of weak stimulation through the TCR. LCK contains N-terminal sites for myristylation and palmitylation, a PTK domain, and SH2 and SH3 domains which are involved in mediating protein-protein interactions with phosphotyrosine-containing and proline-rich motifs, respectively. LCK is also involved in the regulation of apoptosis induced by various stimuli, but not by the death receptors. Diseases associated with LCK include Immunodeficiency 22 and CD45 deficiency. Alternatively splice variants of the LCK gene encoding different isoforms of the protein have been found.

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

Product References

Robust fluorescent detection of protein fatty-acylation with chemical reporters.

Journal of the American Chemical Society

Charron G,Zhang MM,Yount JS,Wilson J,Raghavan AS,Shamir E,Hang HC

MA5-12303 was used in immunoprecipitation and western blot to introduce a new method of fluorescent detection of protein fatty-acylation

Wed Apr 08 00:00:00 EDT 2009

Bioinformatics

Protein Aliases: EC 2.7.10.2; kinase Lck; Leukocyte C-terminal Src kinase; LSK; Lymphocyte cell-specific protein-tyrosine kinase; lymphocyte protein tyrosine kinase; lymphocyte-specific protein tyrosine kinase; OTTHUMP00000008640; OTTHUMP00000008740; OTTHUMP00000008741; p56(LSTRA) protein-tyrosine kinase; p56-LCK; Protein YT16; Proto-oncogene Lck; Proto-oncogene tyrosine-protein kinase LCK; RP4-675E8.4; T cell-specific protein-tyrosine kinase; T-lymphocyte specific protein tyrosine kinase p56lck; tyr; Tyrosine-protein kinase Lck

Gene Aliases: Hck-3; IMD22; LCK; Lck1; Lcktkr; LSK; Lsk-t; Lskt; p56; p56Lck; pp58lck; YT16

UniProt ID: (Human) Q5TDH9, (Rat) Q01621, (Mouse) P06240

Entrez Gene ID: (Human) 3932, (Rat) 313050, (Mouse) 16818

Function(s)

protein phosphorylation cellular zinc ion homeostasis apoptotic process activation of cysteine-type endopeptidase activity involved in apoptotic process cell surface receptor signaling pathway transmembrane receptor protein tyrosine kinase signaling pathway aging response to mechanical stimulus response to metal ion response to zinc ion positive regulation of gene expression dephosphorylation peptidyl-tyrosine phosphorylation T cell differentiation peptidyl-tyrosine autophosphorylation regulation of cell proliferation response to drug positive regulation of tyrosine phosphorylation of Stat5 protein response to hydrogen peroxide regulation of apoptotic process innate immune response positive regulation of gamma-delta T cell differentiation protein autophosphorylation B cell receptor signaling pathway regulation of T cell receptor signaling pathway positive regulation of T cell receptor signaling pathway positive regulation of T cell activation release of sequestered calcium ion into cytosol positive regulation of uterine smooth muscle contraction cellular response to peptide hormone stimulus positive regulation of intrinsic apoptotic signaling pathway phosphorylation protein dephosphorylation regulation of phosphatidylinositol 3-kinase signaling viral process hemopoiesis platelet activation T cell costimulation phosphatidylinositol phosphorylation phosphatidylinositol-mediated signaling regulation of defense response to virus by virus T cell receptor signaling pathway leukocyte migration regulation of lymphocyte activation

Process(es)

glycoprotein binding protein tyrosine kinase activity non-membrane spanning protein tyrosine kinase activity protein serine/threonine phosphatase activity protein binding ATP binding protein C-terminus binding kinase activity protein kinase binding protein phosphatase binding SH2 domain binding CD4 receptor binding CD8 receptor binding identical protein binding phosphatidylinositol 3-kinase binding phosphatidylinositol-4,5-bisphosphate 3-kinase activity ATPase binding antigen binding receptor binding protein complex binding nucleotide binding protein kinase activity transferase activity transferase activity, transferring phosphorus-containing groups

LCK Monoclonal Antibody (3A5)

Product Details

MA5-12303

Western Blot (WB)

Immunocytochemistry (ICC/IF)

Flow Cytometry (Flow)

Immunoprecipitation (IP)

Miscellaneous PubMed (Misc)

Species Reactivity

Published species

Host/Isotype

Class

Type

Clone

Immunogen

Conjugate

Form

Concentration

Purification

Storage buffer

Contains

Storage conditions

RRID

Product Specific Information

Target Information

Bioinformatics

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