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

IgG

Human,Mouse,Rat (predicted:Chicken,Dog,Pig,Cow,Sheep)

1. Tissue/cell: rat brain tissue; 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-C-Jun Polyclonal Antibody, Unconjugated (TMAB-01011) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAb staining.
2. Sample:
Liver (Rat) lysate at 30 μg;
Brain (Rat) lysates at 30 μg;
Primary: Anti-C-jun/AP-1 (TMAB-01011) at 1:200;
Secondary: HRP conjugated Goat-Anti-Rabbit Igg (secondary antibody) at 1: 3000;
Predicted band size: 36 kDa
Observed band size: 36 kDa
3. Paraformaldehyde-fixed, paraffin embedded (Mouse brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (C-jun) Polyclonal Antibody, Unconjugated (TMAB-01011) at 1:500 overnight at 4°C, followed by a conjugated secondary for 20 min and DAB staining.
4. Paraformaldehyde-fixed, paraffin embedded (Rat testis); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (C-jun) Polyclonal Antibody, Unconjugated (TMAB-01011) at 1:400 overnight at 4°C, followed by a conjugated secondary for 20 min and DAB staining.
5. Blank control (blue line): HepG2 (blue).
Primary Antibody (green line): Rabbit Anti-C-jun antibody
Dilution: 1 μg/10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody (white blue line): Goat anti-rabbit IgG-PE
Dilution: 1 μg/test.
Protocol
The cells were fixed with 70% methanol (Overnight at 4°C) and then permeabilized with 90% ice-cold methanol for 20 min at-20°C. Cells stained with Primary Antibody for 30 min at room temperature. The cells were then incubated in 1 X PBS/2% BSA/10% goat serum to block non-specific protein-protein interactions followed by the antibody for 15 min at room temperature. The secondary antibody used for 40 min at room temperature.
6. Paraformaldehyde-fixed, paraffin embedded (Mouse brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (C-jun) Polyclonal Antibody, Unconjugated (TMAB-01011) at 1:400 overnight at 4°C, followed by a conjugated secondary antibody for 20 min and DAB staining.
7. Sample:
Lane 1: Kidney (Mouse) Lysate at 40 μg
Lane 2: Lung (Mouse) Lysate at 40 μg
Lane 3: NIH/3T3 (Mouse) Cell Lysate at 30 μg
Lane 4: Hela (Human) Cell Lysate at 30 μg
Primary: Anti-C-jun (TMAB-01011) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 43/36 kDa
Observed band size: 45 kDa
8. Blank control: Hela.
Primary Antibody (green line): Rabbit Anti-C-jun antibody (TMAB-01011)
Dilution: 1 μg/10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody: Goat anti-rabbit IgG-AF647
Dilution: 1 μg/test.
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: 1μg/Test

Polyclonal

KLH conjugated synthetic peptide: human Transcription factor AP-1

Human

Transcription factors,SMADs,Leucine Zipper,Transcription Factors,TLR Signaling,Phosphoprotein ELISA kits,SMADs