JAK3 Antibody [M5G12] | Primary Antibodies

Application: Reactivity: Human

Lane 1: HEL

Usage Information

Dilution
WB1:2000 IP1:60 IHC1:2000 IF1:100 FCM1:100

Application
WB, IP, IHC, IF, FCM

Reactivity
Human

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 Observed MW
125 kDa 125 kDa
^*Why do the predicted and actual molecular weights differ? The following reasons may explain differences between the predicted and actual protein molecular weight.

Positive Control	Human NK cell lymphoma tissue; Human large B cell lymphoma tissue; TF-1 cells; KARPAS-299 cells; HEL cells; Jurkat cells
Negative Control	HaCaT cells

Datasheet & SDS

Biological Description

Specificity
JAK3 Antibody [M5G12] detects endogenous levels of total JAK3 protein.

Clone
M5G12

Synonym(s)
Tyrosine-protein kinase JAK3; Janus kinase 3; Leukocyte janus kinase; JAK-3; L-JAK; JAK3

Background
JAK3 is a hematopoietic-specific Janus kinase that plays a vital role in cytokine signaling mediated by γc-chain receptors such as those for IL-2, IL-4, IL-7, IL-9, IL-15, and IL-21. It is constitutively associated with the intracellular domains of these receptors and, upon cytokine stimulation, undergoes transphosphorylation that activates its C-terminal kinase domain (JH1), while its N-terminal pseudokinase domain (JH2) provides autoinhibitory regulation through control of the activation loop. Ligand binding triggers JAK3 to autophosphorylate receptor tyrosines, creating SH2-docking sites for STAT proteins (STAT5, STAT3, STAT1), which JAK3 phosphorylates to initiate their dimerization, nuclear entry, and transcriptional activation of genes governing T, B, and NK cell development, survival, and differentiation, including Th1 and Th17 lineages. Mutations in the JH2 domain, such as V722I and G825A, can disrupt autoinhibition and result in gain-of-function activity leading to acute lymphoblastic leukemia, while deletions in the SH2-JH1 linker region also increase basal kinase activity. JAK3 loss-of-function mutations cause severe combined immunodeficiency (SCID) due to failed lymphopoiesis, whereas somatic gain-of-function mutations drive malignancies such as T-cell acute lymphoblastic leukemia, acute myeloid leukemia, and large granular lymphocytic leukemia via persistent STAT activation.

Background

JAK3 is a hematopoietic-specific Janus kinase that plays a vital role in cytokine signaling mediated by γc-chain receptors such as those for IL-2, IL-4, IL-7, IL-9, IL-15, and IL-21. It is constitutively associated with the intracellular domains of these receptors and, upon cytokine stimulation, undergoes transphosphorylation that activates its C-terminal kinase domain (JH1), while its N-terminal pseudokinase domain (JH2) provides autoinhibitory regulation through control of the activation loop. Ligand binding triggers JAK3 to autophosphorylate receptor tyrosines, creating SH2-docking sites for STAT proteins (STAT5, STAT3, STAT1), which JAK3 phosphorylates to initiate their dimerization, nuclear entry, and transcriptional activation of genes governing T, B, and NK cell development, survival, and differentiation, including Th1 and Th17 lineages. Mutations in the JH2 domain, such as V722I and G825A, can disrupt autoinhibition and result in gain-of-function activity leading to acute lymphoblastic leukemia, while deletions in the SH2-JH1 linker region also increase basal kinase activity. JAK3 loss-of-function mutations cause severe combined immunodeficiency (SCID) due to failed lymphopoiesis, whereas somatic gain-of-function mutations drive malignancies such as T-cell acute lymphoblastic leukemia, acute myeloid leukemia, and large granular lymphocytic leukemia via persistent STAT activation.

References
https://pubmed.ncbi.nlm.nih.gov/38474223/ https://pubmed.ncbi.nlm.nih.gov/22130498/

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%