LCK antibody (pTyr505) (Alexa Fluor 647)

Catalog No. ABIN1177084

This Mouse Monoclonal antibody specifically detects LCK in ICS. It exhibits reactivity toward Human, Mouse and Rat.

Quick Overview for LCK antibody (pTyr505) (Alexa Fluor 647) (ABIN1177084)

Target: LCK (Lymphocyte-Specific Protein tyrosine Kinase (LCK))

Reactivity: Human, Mouse, Rat

Host: Mouse

Clonality: Monoclonal

Conjugate: This LCK antibody is conjugated to Alexa Fluor 647

Application: Intracellular Staining (ICS)

Clone: 4-LCK

Product Details anti-LCK Antibody (Alexa Fluor 647)

Binding Specificity: pTyr505

Brand: BD Phosflow™

Purification: The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.

Immunogen: Phosphorylated Human Lck Peptide

Isotype: IgG1

Application Details

Application Notes: This antibody conjugate is suitable for intracellular staining of human whole blood (using BD Phosflow™ Lyse/Fix Buffer) and peripheral blood mononuclear cells (using BD Cytofix™ Fixation Buffer or BD Phosflow™ Fix Buffer I). Any of the three BD Phosflow™ permeabilization buffers may be used.

Sample Volume: 20 μL

Restrictions: For Research Use only

Handling

Format: Liquid

Buffer: Aqueous buffered solution containing BSA and ≤0.09 % sodium azide.

Preservative: Sodium azide

Precaution of Use: This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Storage: 4 °C

Storage Comment: Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The antibody was conjugated to Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was removed.

Target Details for LCK

Target: LCK (Lymphocyte-Specific Protein tyrosine Kinase (LCK))

Alternative Name: Lck

Background: Lck is a member of the Src family of cytoplasmic protein-tyrosine kinases (PTKs) that is normally expressed exclusively in lymphoid cells, primarily T lymphocytes and NK cells. Members of this family have several common features: 1) unique N-terminal domains, 2) attachment to cellular membranes through a myristylated N-terminus, and 3) homologous SH2, SH3, and catalytic domains. The unique N-terminal domain of Lck interacts with the cytoplasmic tails of the CD4 and CD8 cell-surface glycoproteins of T lymphocytes, which recognize antigen presenting cells via their surface MHC class II and class I molecules, respectively. The catalytic activity of Lck is regulated by both kinases and phosphatases that control the phosphorylation states of two tyrosine residues that have opposing effects. Repression of Lck's catalytic activity occurs via phosphorylation at tyrosine 505 (Y505), located near the carboxy terminus. Phosphorylation of this tyrosine site is mediated by the Csk family of PTKs, and its dephosphorylation is mediated by the protein tyrosine phosphatase, CD45. When Lck is phosphorylated at this site, it assumes a folded tertiary structure which is enzymatically inactive. When CD45 dephosphorylates it at Y505, Lck is able to autophosphorylate its Y394, which leads to conformational changes in the catalytic domain that induce kinase activity. However, it has been observed that the inhibitory effect of the phosphorylated Y505 can be overcome by direct engagement of Lck's SH3 domain and that both Y394 and Y505 are phosphorylated together in cells activated by hydrogen peroxide. Activated Lck phosphorylates the ITAMs (Immunoreceptor-based Tyrosine Activation Motifs) of the T cell receptor (TCR) and thus is critical for activation and development of T lymphocytes. The interactions of Lck, Csk, CD45, CD4 or CD8, and TCR are only a small part of a complex immunoregulatory cascade that involves additional substrates for Csk and CD45, other enzymes, adhesion molecules, adaptor proteins, and specialized membrane microdomains. The 4/LCK-Y505 monoclonal antibody recognizes the phosphorylated Y505 of the catalytic domain of Lck. The Alexa Fluor® 488- conjugated format has been evaluated by flow using a human model system. However, the unconjugated form of this antibody (Cat. No. 612390) has been shown to react with human, mouse, and rat in western blot. A phosphorylated peptide corresponding to residues around Tyrosine-505 from human Lck was used as the immunogen.

Pathways: TCR Signaling, Fc-epsilon Receptor Signaling Pathway, EGFR Signaling Pathway, Neurotrophin Signaling Pathway, Transition Metal Ion Homeostasis, Positive Regulation of Endopeptidase Activity, CXCR4-mediated Signaling Events, Thromboxane A2 Receptor Signaling