Anti-GJA1 Polyclonal Antibody is a Rabbit antibody targeting GJA1. Anti-GJA1 Polyclonal Antibody can be used in WB,IHC-P,IHC-Fr,ICC/IF,IF,FCM.

IgG

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

1. Paraformaldehyde-fixed, paraffin embedded (Human stomach cancer); 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 (Connexin 43) Polyclonal Antibody, Unconjugated (TMAB-00772) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.
2. Tissue/cell: rat heart tissue;4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer (0.01M, pH6.0), Boiling bathing for 15 min; Blocking buffer (normal goat serum) at 37°C for 20 min;
Incubation: Anti-Connexin 43 Polyclonal Antibody, Unconjugated (TMAB-00772) 1:200, overnight at 4°C; The secondary antibody was Goat Anti-Rabbit IgG, PE conjugated used at 1:200 dilution for 40 minutes at 37°C.
3. Sample: Mcf-7 Cell Lysate at 40 μg
Primary: Anti-Connexin (TMAB-00772) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/10000 dilution
Predicted band size: 42 kDa
Observed band size: 43 kDa
4. Tissue/cell: MCF7 cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum) at 37°C for 20 min; Antibody incubation with (Connexin 43) polyclonal Antibody, Unconjugated (TMAB-00772) 1:100, 90 minutes at 37°C; followed by a FITC conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nucleus.
5. Tissue/cell: U-251 cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum) at 37°C for 20 min; Antibody incubation with (Connexin 43) polyclonal Antibody, Unconjugated (TMAB-00772) 1:100, 90 minutes at 37°C; followed by a FITC conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nucleus.
6. Sample:
Lane 1: Heart (Mouse) Lysate at 40 μg
Lane 2: Heart (Rat) Lysate at 40 μg
Lane 3: Cerebrum (Mouse) Lysate at 40 μg
Lane 4: Cerebrum (Rat) Lysate at 40 μg
Primary: Anti-Connexin 43 (TMAB-00772) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 42 kDa
Observed band size: 45 kDa
7. Blank control (blue line): Hela (blue).
Primary Antibody (green line): Rabbit Anti-Connexin 43 antibody (TMAB-00772)
Dilution: 1 μg/10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody (white blue line): f (ab/)2 fragment goat anti-rabbit IgG-FITC.
Dilution: 1 μg/test.
Protocol
The cells were fixed with 2% paraformaldehyde (10 min), then permeabilized with 90% ice-cold methanol for 30 min on ice. 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.
8. 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 (Connexin 43) Polyclonal Antibody, Unconjugated (TMAB-00772) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

WB: 1:500-2000; IHC-P: 1:100-500; IHC-Fr: 1:100-500; ICC/IF: 1:100; IF: 1:100-500; FCM: 1μg/Test

Polyclonal

KLH conjugated synthetic peptide: human Connexin-43

Human

Cell junction molecules,Cardiac arrhythmias,Gap Junctions