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,IHC-P,IHC-Fr,IF,FCM

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

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