Phospho-LYN (Y397)/LCK (Y394)/HCK (Y411)/BLK (Y389) Antibody [G2C4]

Application: Reactivity: Human, Mouse

Lane 1: K562, Lane 2: K562 (CIP treated)

Usage Information

Dilution
WB1:1000 IP1:100 FCM1:200-1:800

Application
WB, IP, FCM

Reactivity
Human, Mouse

Source
Rabbit Monoclonal Antibody

Storage Buffer
PBS, pH 7.2+50% Glycerol+0.05% BSA+0.01% NaN3

Storage (from the date of receipt)
-20°C (avoid freeze-thaw cycles), 2 years

Predicted MW
60 kDa

Datasheet & SDS

Biological Description

Specificity
Phospho-LYN (Y397)/LCK (Y394)/HCK (Y411)/BLK (Y389) Antibody [G2C4] detects endogenous levels of total LYN, LCK, HCK, and BLK protein only when it is phosphorylated at Y397, Y394, Y411, and Y389 respectively.

Clone
G2C4

Synonym(s)
Tyrosine-protein kinase Lyn/Lck/Hck/Blk; LYN; LCK; HCK; BLK

Background
Phospho-LYN (Y397)/LCK (Y394)/HCK (Y411)/BLK (Y389) represent the activated forms of Src family kinases (SFKs) that transmit signals from immune receptors through autophosphorylation in their kinase activation loops. These kinases share a conserved SH3-SH2-kinase domain architecture where phosphorylation at homologous tyrosine residues, corresponding to Y416 in c-Src, displaces the activation loop from the catalytic cleft, enabling ATP binding and substrate access while relieving autoinhibitory SH2 clamping by C-terminal tyrosine phosphorylation. Receptor ligation triggers trans-autophosphorylation: LCK Y394 activation upon TCR/CD3 clustering recruits ZAP-70 via tandem ITAM motifs, propagating PLCγ-IP3-Ca2+ flux and NFAT nuclear translocation for T cell effector differentiation; LYN Y397 engages BCR/CD79a/b in B cells to phosphorylate SYK and activate PI3K-AKT-mTOR for survival while balancing ITIM-mediated inhibition through CD22/SHP-1 recruitment; HCK Y411 drives FcγR and integrin signaling in myeloid phagocytes via SYK-Vav-Rac cascades that reorganize actin for podosome formation, NADPH oxidase assembly, and cytokine release; BLK Y389 supports pre-BCR checkpoints through BLNK-SYK coupling that enforces light chain selection during B lymphopoiesis. Dual phosphorylation dynamics, activation loop versus C-terminal tail, integrate positive (CD28, CD40L) and negative (CD45, Csk) regulation to calibrate immune synapse maturation and prevent autoimmunity. These phospho-SFKs govern thymic selection, marginal zone B cell positioning, and macrophage polarization with tissue-specific expression patterns reflecting lineage commitment.

Background

Phospho-LYN (Y397)/LCK (Y394)/HCK (Y411)/BLK (Y389) represent the activated forms of Src family kinases (SFKs) that transmit signals from immune receptors through autophosphorylation in their kinase activation loops. These kinases share a conserved SH3-SH2-kinase domain architecture where phosphorylation at homologous tyrosine residues, corresponding to Y416 in c-Src, displaces the activation loop from the catalytic cleft, enabling ATP binding and substrate access while relieving autoinhibitory SH2 clamping by C-terminal tyrosine phosphorylation. Receptor ligation triggers trans-autophosphorylation: LCK Y394 activation upon TCR/CD3 clustering recruits ZAP-70 via tandem ITAM motifs, propagating PLCγ-IP3-Ca2+ flux and NFAT nuclear translocation for T cell effector differentiation; LYN Y397 engages BCR/CD79a/b in B cells to phosphorylate SYK and activate PI3K-AKT-mTOR for survival while balancing ITIM-mediated inhibition through CD22/SHP-1 recruitment; HCK Y411 drives FcγR and integrin signaling in myeloid phagocytes via SYK-Vav-Rac cascades that reorganize actin for podosome formation, NADPH oxidase assembly, and cytokine release; BLK Y389 supports pre-BCR checkpoints through BLNK-SYK coupling that enforces light chain selection during B lymphopoiesis. Dual phosphorylation dynamics, activation loop versus C-terminal tail, integrate positive (CD28, CD40L) and negative (CD45, Csk) regulation to calibrate immune synapse maturation and prevent autoimmunity. These phospho-SFKs govern thymic selection, marginal zone B cell positioning, and macrophage polarization with tissue-specific expression patterns reflecting lineage commitment.

References
https://pubmed.ncbi.nlm.nih.gov/22805580/ https://pubmed.ncbi.nlm.nih.gov/28096507/

Reference Table for Selecting PVDF Membrane Pore Size Specifications

Protein Size (kDa)	PVDF Transfer Membrane Pore Size (μm)
< 10	0.22
10-20	0.22
20-30	0.45
30-45	0.45
45-70	0.45
80-100	0.45
100-140	0.45
> 140	0.45

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Protein Size (kDa)	Separating Gel Percentage
4-40	20%
12-45	15%
10-70	12.5%
15-100	10%
25-100	8%
60-210	5%