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Human Phospho-Lck (Y394) Antibody

R&D Systems, part of Bio-Techne | Catalog # MAB7500

R&D Systems, part of Bio-Techne
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MAB7500
MAB7500-SP

Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human

Cited:

Human, Mouse

Applications

Validated:

Immunocytochemistry, Western Blot

Cited:

Immunocytochemistry, Western Blot

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG2B Clone # 755103

Product Specifications

Immunogen

Phosphopeptide containing the human Lck Y394 site
Accession # P06239

Specificity

Detects human Lck when phosphorylated at Y394. Due to high sequence homology, this antibody may cross-react with phosphorylated members of the Src family.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2B

Scientific Data Images for Human Phospho-Lck (Y394) Antibody

Detection of Human Phospho-Lck (Y394) antibody by Western Blot.

Detection of Human Phospho-Lck (Y394) by Western Blot.

Western blot shows lysates of Jurkat human acute T cell leukemia cell line and Ramos human Burkitt's lymphoma cell line untreated (-) or treated (+) with 1 mM Pervanadate (PV) for 5 minutes. PVDF membrane was probed with 0.1 µg/mL of Mouse Anti-Human Phospho-Lck (Y394) Monoclonal Antibody (Catalog # MAB7500) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for Phospho-Lck (Y394) at approximately 56 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Phospho-Lck (Y394) antibody in Jurkat Human Cell Line by Immunocytochemistry (ICC).

Phospho‑Lck (Y394) in Jurkat Human Cell Line.

Lck phosphorylated at Y394 was detected in immersion fixed Jurkat human acute T cell leukemia cells treated with (upper panel) or without (lower panel) Pervanadate using Mouse Anti-Human Phospho-Lck (Y394) Monoclonal Antibody (Catalog # MAB7500) 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.

Applications for Human Phospho-Lck (Y394) Antibody

Application
Recommended Usage

Immunocytochemistry

8-25 µg/mL
Sample: Immersion fixed Jurkat human acute T cell leukemia cell line treated with Pervanadate

Western Blot

0.1 µg/mL
Sample: Jurkat human acute T cell leukemia cell line and Ramos human Burkitt's lymphoma cell line treated with Pervanadate
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Sterile PBS to a final concentration of 0.5 mg/mL. For liquid material, refer to CoA for concentration.

Reconstitution Buffer Available:
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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.

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). 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. Y394 is the major site for autophosphorylation that enhances kinase activity, while Y505 is the major phosphorylation site for negative regulation.

Long Name

Lymphocyte Protein Tyrosine Kinase

Alternate Names

Hck-3, Lcktkr, LSTRA, p56lck, pp58lck, YT16

Entrez Gene IDs

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

Gene Symbol

LCK

UniProt

Additional Lck Products

Product Documents for Human Phospho-Lck (Y394) Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human Phospho-Lck (Y394) Antibody

For research use only

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