Human Phospho-Lck (Y505) Antibody

Catalog #: MAB7369 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
MAB7369
MAB7369-SP

Detection of Human Phospho-Lck (Y505) by Western Blot.

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Human Phospho-Lck (Y505) Antibody Summary

Species Reactivity

Human

Specificity

Detects human Phospho-Lck (Y505) in direct ELISA and Western Blot.

Source

Monoclonal Mouse IgG_2A Clone # 755216

Purification

Protein A or G purified from hybridoma culture supernatant

Immunogen

Phosphopeptide containing the human Lck (Y505) site.
Accession # P06239

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Applications

Recommended Concentration

Sample

Western Blot

0.5 µg/mL

See below

Immunocytochemistry

8-25 µg/mL

See below

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Western Blot

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Detection of Human Phospho-Lck (Y505) by Western Blot. Western blot shows lysates of Jurkat human acute T cell leukemia cell line untreated (-) or treated (+) with 1 mM Pervanadate (PV) for 10 minutes or 2mM H₂O₂for 2 minutes. PVDF membrane was probed with 0.5 µg/mL of Mouse Anti-Human Phospho-Lck (Y505) Monoclonal Antibody (Catalog # MAB7369) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for Phospho-Lck (Y505) at approximately 62 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Immunocytochemistry

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Phospho-Lck (Y505) in Jurkat Human Cell Line. Lck phosphorylated at Y505 was detected in immersion fixed Jurkat human acute T cell leukemia cell line stimulated (upper panel) or not (lower panel) with pervanadate using Mouse Anti-Human Phospho-Lck (Y505) Monoclonal Antibody (Catalog # MAB7369) 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 cell surfaces and cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Sterile PBS to a final concentration of 0.5 mg/mL.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: Lck

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.

Long Name

Lymphocyte Protein Tyrosine Kinase

Entrez Gene IDs

3932 (Human); 16818 (Mouse); 313050 (Rat)

Alternate Names

EC 2.7.10; EC 2.7.10.2; Hck-3; Lck; Lcktkr; Leukocyte C-terminal Src kinase; LSK; LSTRA; Lymphocyte cell-specific protein-tyrosine kinase; lymphocyte-specific protein tyrosine kinase; p56(LSTRA) protein-tyrosine kinase; p56lck; p56-LCK; pp58lck; Protein YT16; Proto-oncogene Lck; proto-oncogene tyrosine-protein kinase LCK; T cell-specific protein-tyrosine kinase; T-lymphocyte specific protein tyrosine kinase p56lck; tyrosine-protein kinase Lck; YT16