Alpha smooth muscle Actin Recombinant Rabbit Monoclonal Antibody

• Catalog Number: orb704355
• Category: Antibodies
• Description: Alpha smooth muscle Actin Recombinant Rabbit Monoclonal Antibody
• Species/Host: Rabbit
• Clonality: Recombinant
• Clone Number: 2B10
• Tested applications: FC, ICC, IF, IHC-Fr, IHC-P, WB
• Predicted Reactivity: Rat
• Reactivity: Human, Mouse, Rat
• Isotype: IgG
• Immunogen: KLH conjugated synthetic peptide derived from human Actin alpha
• Concentration: 1mg/ml
• Dilution range: WB=1:1000-2000, IHC-P=1:50-200, IHC-F=1:50-200, ICC/IF=1:50, IF=1:50-200, Flow-Cyt=1:50
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: 42 kDa
• Target: ACTA2
• UniProt ID: P62736
• 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: alpha sarcomeric Actin; alpha smooth muscle Actin;
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

A431 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 (alpha smooth muscle Actin) Monoclonal Antibody, Unconjugated (orb704355) 1:50, 90 minutes at 37°C, followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nuclei.

A549 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 (alpha smooth muscle Actin) Monoclonal Antibody, Unconjugated (orb704355) 1:50, 90 minutes at 37°C, followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nuclei.

Blank control: Jurkat. Primary Antibody (green line): Rabbit Anti-alpha smooth muscle Actin antibody (orb704355), dilution: 1:50, Isotype Control Antibody (orange line): Rabbit IgG. Secondary Antibody: Goat anti-rabbit IgG-AF488, dilution: 1:1000. Protocol, The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 0.1% PBST for 20 min at room temperature. 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. Acquisition of 20000 events was performed.

HepG2 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 (alpha smooth muscle Actin) Monoclonal Antibody, Unconjugated (orb704355) 1:50, 90 minutes at 37°C, followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nuclei.

Paraformaldehyde-fixed, paraffin embedded (human liver tissue), 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 (alpha smooth muscle Actin) Monoclonal Antibody, Unconjugated (orb704355) at 1:50 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (human stomach carcinoma tissue), 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 (alpha smooth muscle Actin) Monoclonal Antibody, Unconjugated (orb704355) at 1:50 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (human tonsil tissue), 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 (alpha smooth muscle Actin) Monoclonal Antibody, Unconjugated (orb704355) at 1:50 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

RH-35 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 (alpha smooth muscle Actin) Monoclonal Antibody, Unconjugated (orb704355) 1:50, 90 minutes at 37°C, followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nuclei.

Sample: Lane 1: A431 cell lysate, Lane 2: Hela cell lysate, Lane 3: NIH/3T3 cell lysate, Lane 4: mouse heart tissue lysate, Lane 5: mouse smooth muscle tissue lysate, Primary: Anti-alpha smooth muscle Actin (orb704355) at 1:500 dilution, Secondary: Goat Anti-Rabbit IgG - HRP at 1:5000 dilution, Predicted band size: 42 kD, Observed band size: 42 kD.