Anti-JAK3 Polyclonal Antibody is a Rabbit antibody targeting JAK3. Anti-JAK3 Polyclonal Antibody can be used in WB,IHC-P,IHC-Fr,IF,FCM.

IgG

Human,Mouse,Rat (predicted:Dog,Pig,Cow,Horse)

1. Tissue/cell: human lung carcinoma; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer (0.01M, pH6.0), Boiling bathing for 15 min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min; Blocking buffer (normal goat serum) at 37°C for 20 min;
Incubation: Anti-Jak3 Polyclonal Antibody, Unconjugated (TMAB-01005) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAb staining.
2. Sample:
SP2/0 Cell (Mouse) Lysate at 40 μg
Jurkat Cell (Human) Lysate at 40 μg
K562 Cell (Human) Lysate at 40 μg
Primary: Anti-Jak3 (TMAB-01005) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 125 kDa
Observed band size: 125 kDa
3. Blank control: Molt-4. Primary Antibody (green line): Rabbit Anti-Jak3 antibody (TMAB-01005)
Dilution: 0.2 μg/10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody: Goat anti-rabbit IgG-PE
Dilution: 0.2 μg/test.
Protocol
The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 20% PBST for 20 min at-20°C. The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature.
4. Sample:
Lane 1: Mouse Thymus tissue lysates
Lane 2: Rat LympHnode tissue lysates
Lane 3: Human Jurkat cell lysates
Lane 4: Human Hela cell lysates
Lane 5: Human Molt-4 cell lysates
Primary: Anti-Jak3 (TMAB-01005) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 125 kDa
Observed band size: 115 kDa
5. Blank control (black line): Molt4. Primary Antibody (green line): Rabbit Anti-Jak3 antibody (TMAB-01005)
Dilution: 1 μg/Test;
Secondary Antibody (white blue line): Goat anti-rabbit IgG-AF488
Dilution: 0.5 μg/Test.
Isotype control (orange line): Normal Rabbit IgG
Protocol
The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 90% ice-cold methanol for 20 min at-20°C, The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature.

WB: 1:500-2000; IHC-P: 1:100-500; IHC-Fr: 1:100-500; IF: 1:100-500; FCM: 1ug/test

Polyclonal

KLH conjugated synthetic peptide: human Jak3

Human

JAK3

Tyrosine-protein kinase JAK3

Signal transducers,Tyrosine kinases,STATs,Transcription Factors,Tyrosine Kinases,STATs

Non-receptor tyrosine kinase involved in various processes such as cell growth, development, or differentiation. Mediates essential signaling events in both innate and adaptive immunity and plays a crucial role in hematopoiesis during T-cells development. In the cytoplasm, plays a pivotal role in signal transduction via its association with type I receptors sharing the common subunit gamma such as IL2R, IL4R, IL7R, IL9R, IL15R and IL21R. Following ligand binding to cell surface receptors, phosphorylates specific tyrosine residues on the cytoplasmic tails of the receptor, creating docking sites for STATs proteins. Subsequently, phosphorylates the STATs proteins once they are recruited to the receptor. Phosphorylated STATs then form homodimer or heterodimers and translocate to the nucleus to activate gene transcription. For example, upon IL2R activation by IL2, JAK1 and JAK3 molecules bind to IL2R beta (IL2RB) and gamma chain (IL2RG) subunits inducing the tyrosine phosphorylation of both receptor subunits on their cytoplasmic domain. Then, STAT5A AND STAT5B are recruited, phosphorylated and activated by JAK1 and JAK3. Once activated, dimerized STAT5 translocates to the nucleus and promotes the transcription of specific target genes in a cytokine-specific fashion.