Alpha smooth muscle Actin Rabbit Polyclonal Antibody

• Catalog Number: orb155585
• Category: Antibodies
• Description: Alpha smooth muscle Actin Rabbit Polyclonal Antibody
• Target: ACTA2
• Clonality: Polyclonal
• Species/Host: Rabbit
• Isotype: IgG
• Conjugation: Unconjugated
• Reactivity: Human, Mouse, Rat
• Predicted Reactivity: Rabbit
• Form/Appearance: Liquid
• Concentration: 1mg/ml
• Buffer/Preservatives: 0.01M TBS (pH7.4) with 1% rAlbumin, 0.02% Proclin300 and 50% Glycerol.
• Immunogen: KLH conjugated synthetic peptide derived from human Actin alpha (165-260/377aa)
• UniProt ID: P62736
• MW: 42 kDa
• Tested applications: FC, IF, IHC-Fr, IHC-P, WB
• Dilution range: WB=1:1000-5000, IHC-P=1:100-500, IHC-F=1:100-500, IF=1:100-500, Flow-Cyt=1μg/Test
• Antibody Type: Primary Antibody
• 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.
• Alternative names: alpha sarcomeric Actin; alpha smooth muscle Actin;
• Note: For research use only

Blank control (blue line): Hela (fixed with 70% ethanol (Overnight at 4°C) and then permeabilized with 90% ice-cold methanol for 30 min on ice). Primary Antibody (green line): Rabbit Anti-alpha smooth muscle Actin antibody (orb155585), Dilution: 1 µg/10^6 cells, Isotype Control Antibody (orange line): Rabbit IgG. Secondary Antibody (white blue line): Goat anti-rabbit IgG-FITC, Dilution: 1 µg/Test.

Blank control: NIH/3T3. Primary Antibody (green line): Rabbit Anti-alpha smooth muscle Actin antibody (orb155585), Dilution: 1 µg/10^6 cells, Isotype Control Antibody (orange line): Rabbit IgG. Secondary Antibody: Goat anti-rabbit IgG-AF488, 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. Acquisition of 20000 events was performed.

Paraformaldehyde-fixed, paraffin embedded (human cervical carcinoma), 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) Polyclonal Antibody, Unconjugated (orb155585) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand.

Paraformaldehyde-fixed, paraffin embedded (human colon carcinoma), 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) Polyclonal Antibody, Unconjugated (orb155585) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand.

Paraformaldehyde-fixed, paraffin embedded (Human duodenum), 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) Polyclonal Antibody, Unconjugated (orb155585) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (Human stomach), 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) Polyclonal Antibody, Unconjugated (orb155585) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (mouse stomach), 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) Polyclonal Antibody, Unconjugated (orb155585) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (rat colon), 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) Polyclonal Antibody, Unconjugated (orb155585) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (rat stomach), 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) Polyclonal Antibody, Unconjugated (orb155585) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Sample: A549 (Human) Cell Lysate at 30 ug, NIH/3T3 (Mouse) Cell Lysate at 30 ug, Hela (Human) Cell Lysate at 30 ug, A431 (Human) Cell Lysate at 30 ug, HepG2 (Human) Cell Lysate at 30 ug, Stomach (Mouse) Lysate at 40 ug, Lung (Mouse) Lysate at 40 ug, Skeletal muscle (-) (Mouse) Lysate at 40 ug, Primary: Anti-alpha smooth muscle Actin (orb155585) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 42 kD, Observed band size: 42 kD.

Sample: Liver (Rat) Lysate at 40 ug, Urinary bladder (Rat) Lysate at 40 ug, Stomach (Mouse) Lysate at 40 ug, A549 (Cell) Lysate at 30 ug, Primary: Anti-alpha smooth muscle Actin (orb155585) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 42 kD, Observed band size: 46 kD.