Connexin 43 Rabbit Polyclonal Antibody

• Catalog Number: orb10485
• Category: Antibodies
• Description: Connexin 43 Rabbit Polyclonal Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: FC, ICC, IF, IHC-Fr, IHC-P, WB
• Predicted Reactivity: Bovine, Canine, Gallus
• Reactivity: Human, Mouse, Rat
• Isotype: IgG
• Immunogen: KLH conjugated synthetic peptide derived from human Connexin-43 (211-260/382 humanaa)
• Antibody Type: Primary Antibody
• Concentration: 1mg/ml
• Dilution range: WB=1:500-2000, IHC-P=1:100-500, IHC-F=1:100-500, ICC/IF=1:100, IF=1:100-500, Flow-Cyt=1μg/Test
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: 42 kDa
• Target: GJA1
• UniProt ID: P17302
• RRID: AB_10751264
• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Buffer/Preservatives: 0.01M TBS (pH7.4) with 1% rAlbumin, 0.02% Proclin300 and 50% Glycerol.
• Alternative names: Connexin 43; connexin43; Connexin43v; Cx 43v; CX43
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

Blank control (blue line): Hela (blue). Primary Antibody (green line): Rabbit Anti-Connexin 43 antibody (orb10485), 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. Acquisition of 20000 events was performed.

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 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (Connexin 43) Polyclonal Antibody, Unconjugated (orb10485) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.

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 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (Connexin 43) Polyclonal Antibody, Unconjugated (orb10485) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Sample: Lane 1: Heart (Mouse) Lysate at 40 ug, Lane 2: Heart (Rat) Lysate at 40 ug, Lane 3: Cerebrum (Mouse) Lysate at 40 ug, Lane 4: Cerebrum (Rat) Lysate at 40 ug, Primary: Anti-Connexin 43 (orb10485) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 42 kD, Observed band size: 45 kD.

Sample: Mcf-7 Cell Lysate at 40 ug, Primary: Anti-Connexin (orb10485) at 1/300 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/10000 dilution, Predicted band size: 42 kD, Observed band size: 43 kD.

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 (orb10485) 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 nuclei.

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 (orb10485) 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 nuclei.