Alpha smooth muscle Actin Mouse Monoclonal Antibody

• Catalog Number: orb500841
• Category: Antibodies
• Description: Alpha smooth muscle Actin Mouse Monoclonal Antibody
• Species/Host: Mouse
• Clonality: Monoclonal
• Clone Number: 3F9
• Tested applications: FC, IF, IHC-Fr, IHC-P, WB
• Predicted Reactivity: Gallus
• Reactivity: Human, Mouse, Rat
• Isotype: IgG
• Immunogen: KLH conjugated synthetic peptide derived from human Actin alpha
• Concentration: 1mg/ml
• Dilution range: WB=1:500-5000, IHC-P=1:100-500, IHC-F=1:100-500, IF=1:100-500, Flow-Cyt=1ug/Test
• 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.

Blank control: NIH/3T3. Primary Antibody (green line): Mouse Anti-alpha smooth muscle Actin antibody (orb500841), dilution: 1 ug/Test, Secondary Antibody: Goat anti-mouse IgG-FITC, dilution: 0.5 ug/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 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) Monoclonal Antibody, Unconjugated (ascites of orb500841 3F9) at 1:2000 overnight at 4°C, followed by operating according to SP Kit (Mouse) instructions and DAB staining.

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) Monoclonal Antibody, Unconjugated (orb500841) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Mouse) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (Human uterus), 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 (orb500841) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Mouse) 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) Monoclonal Antibody, Unconjugated (orb500841) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Mouse) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (mouse uterus), 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 (orb500841) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Mouse) 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) Monoclonal Antibody, Unconjugated (ascites of orb500841 3F9) at 1:2000 overnight at 4°C, followed by operating according to SP Kit (Mouse) instructions and 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) Monoclonal Antibody, Unconjugated (orb500841) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Mouse) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (rat uterus), 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 (orb500841) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Mouse) instructionsand DAB staining.

Sample: Lane 1: Large intestine (Mouse) Lysate at 40 ug, Lane 2: Stomach (Mouse) Lysate at 40 ug, Lane 3: Uterus (Mouse) Lysate at 40 ug, Lane 4: Kidney (Mouse) Lysate at 40 ug, Lane 5: Small intestine (Mouse) Lysate at 40 ug, Lane 6: NIH/3T3 (Mouse) Cell Lysate at 30 ug, Lane 7: Stomach (Rat) Lysate at 40 ug, Lane 8: Uterus (Rat) Lysate at 40 ug, Lane 9: A549 (Human) Cell Lysate at 30 ug, Lane 10: Hela (Human) Cell Lysate at 30 ug, Lane 11: A431 (Human) Cell Lysate at 30 ug, Lane 12: HepG2 (Human) Cell Lysate at 30 ug, Primary: Anti-alpha smooth muscle Actin (orb500841) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Mouse IgG at 1/20000 dilution, Predicted band size: 42 kD, Observed band size: 42 kD.