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    products/primary-antibodies/alpha-smooth-muscle-actin-antibody-sp171-ab150301.pdf

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Signal Transduction Cytoskeleton / ECM Cytoskeleton Microfilaments Actin etc Actin
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-alpha smooth muscle Actin antibody [SP171] (ab150301)

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  • SDS
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Immunocytochemistry/ Immunofluorescence - Anti-alpha smooth muscle Actin antibody [SP171] (ab150301)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody [SP171] (ab150301)
  • Immunocytochemistry/ Immunofluorescence - Anti-alpha smooth muscle Actin antibody [SP171] (ab150301)
  • Western blot - Anti-alpha smooth muscle Actin antibody [SP171] (ab150301)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody [SP171] (ab150301)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody [SP171] (ab150301)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody [SP171] (ab150301)
  • Immunocytochemistry/ Immunofluorescence - Anti-alpha smooth muscle Actin antibody [SP171] (ab150301)
  • Western blot - Anti-alpha smooth muscle Actin antibody [SP171] (ab150301)
  • Flow Cytometry (Intracellular) - Anti-alpha smooth muscle Actin antibody [SP171] (ab150301)
  • Flow Cytometry (Intracellular) - Anti-alpha smooth muscle Actin antibody [SP171] (ab150301)
  • Flow Cytometry - Anti-alpha smooth muscle Actin antibody [SP171] (ab150301)
  • Anti-alpha smooth muscle Actin antibody [SP171] (ab150301)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [SP171] to alpha smooth muscle Actin
  • Suitable for: Flow Cyt, ICC/IF, WB, IHC-P
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 647 Carrier Free

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Overview

  • Product name

    Anti-alpha smooth muscle Actin antibody [SP171]
    See all alpha smooth muscle Actin primary antibodies
  • Description

    Rabbit monoclonal [SP171] to alpha smooth muscle Actin
  • Host species

    Rabbit
  • Tested applications

    Suitable for: Flow Cyt, ICC/IF, WB, IHC-Pmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
    Predicted to work with: Rabbit, Chicken, Cow, Pig
  • Immunogen

    Synthetic peptide within Human alpha smooth muscle Actin aa 1-100 (N terminal). The exact sequence is proprietary.
    Database link: P62736

  • Positive control

    • IHC-P: Human colon, Mouse colon, and Rat colon tissue. WB: Recombinant Human alpha smooth muscle Actin protein (ab114148); HeLa cell lysate. Flow Cyt: HeLa, NIH/3T3 and C6 cells. ICC/IF: NIH/3T3, SV40LT-SMC, and HeLa cells.
  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    This product is FOR RESEARCH USE ONLY. For commercial use, please contact partnerships@abcam.com.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C. Do Not Freeze.
  • Storage buffer

    pH: 7.60
    Preservative: 0.1% Sodium azide
    Constituents: PBS, 1% BSA
  • Concentration information loading...
  • Purity

    Protein A/G purified
  • Purification notes

    Purified from TCS by protein A/G.
  • Clonality

    Monoclonal
  • Clone number

    SP171
  • Isotype

    IgG
  • Research areas

    • Signal Transduction
    • Cytoskeleton / ECM
    • Cytoskeleton
    • Microfilaments
    • Actin etc
    • Actin
    • Stem Cells
    • Lineage Markers
    • Mesoderm
    • Stem Cells
    • Mesenchymal Stem Cells
    • Myogenesis
    • Cardiovascular
    • Heart
    • Contractility
    • Contractile Proteins
    • Actins
    • Cardiovascular
    • Vasculature
    • Vasculature Markers
    • Arterial
    • Cardiovascular
    • Vasculature
    • Smooth muscle cell (SMC)
    • Cytoskeleton
    • Developmental Biology
    • Lineage specification
    • Mesoderm
    • Cardiovascular
    • Cardiovascular Markers
    • Cell Markers
    • Smooth Muscle Cells

Associated products

  • Alternative Versions

    • Anti-alpha smooth muscle Actin antibody [SP171] - BSA and Azide free (ab242395)
    • Alexa Fluor® 488 Anti-alpha smooth muscle Actin antibody [SP171] (ab267536)
    • Alexa Fluor® 647 Anti-alpha smooth muscle Actin antibody [SP171] (ab267537)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081)
    • Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human ACTA2 knockout HeLa cell line (ab264014)
  • KO cell lysates

    • Human ACTA2 knockout HeLa cell lysate (ab264499)
  • Positive Controls

    • Recombinant Human alpha smooth muscle Actin protein (ab134555)
    • HeLa whole cell lysate (ab150035)
    • HeLa whole cell lysate (ab29545)
  • Recombinant Protein

    • Recombinant Human alpha smooth muscle Actin protein (ab134555)
  • Related Products

    • Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab150301 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Flow Cyt
Use at an assay dependent concentration.
ICC/IF
Use a concentration of 1 - 5 µg/ml.
WB
1/50. Predicted molecular weight: 42 kDa.
IHC-P
1/200.
Notes
Flow Cyt
Use at an assay dependent concentration.
ICC/IF
Use a concentration of 1 - 5 µg/ml.
WB
1/50. Predicted molecular weight: 42 kDa.
IHC-P
1/200.

Target

  • Function

    Actins are highly conserved proteins that are involved in various types of cell motility and are ubiquitously expressed in all eukaryotic cells.
  • Involvement in disease

    Defects in ACTA2 are the cause of aortic aneurysm familial thoracic type 6 (AAT6) [MIM:611788]. AATs are characterized by permanent dilation of the thoracic aorta usually due to degenerative changes in the aortic wall. They are primarily associated with a characteristic histologic appearance known as 'medial necrosis' or 'Erdheim cystic medial necrosis' in which there is degeneration and fragmentation of elastic fibers, loss of smooth muscle cells, and an accumulation of basophilic ground substance.
  • Sequence similarities

    Belongs to the actin family.
  • Cellular localization

    Cytoplasm > cytoskeleton.
  • Target information above from: UniProt accession P62736 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 423787 Chicken
    • Entrez Gene: 515610 Cow
    • Entrez Gene: 59 Human
    • Entrez Gene: 11475 Mouse
    • Entrez Gene: 733615 Pig
    • Entrez Gene: 100009271 Rabbit
    • Entrez Gene: 81633 Rat
    • Omim: 102620 Human
    • SwissProt: P08023 Chicken
    • SwissProt: P62739 Cow
    • SwissProt: P62736 Human
    • SwissProt: P62737 Mouse
    • SwissProt: P62740 Rabbit
    • SwissProt: P62738 Rat
    • Unigene: 500483 Human
    • Unigene: 213025 Mouse
    • Unigene: 195319 Rat
    • Unigene: 3114 Rat
    see all
  • Alternative names

    • a actin antibody
    • AAT6 antibody
    • ACTA_HUMAN antibody
    • ACTA2 antibody
    • Actin alpha 2 smooth muscle aorta antibody
    • Actin aortic smooth muscle antibody
    • Actin, aortic smooth muscle antibody
    • ACTSA antibody
    • ACTVS antibody
    • Alpha 2 actin antibody
    • Alpha actin 2 antibody
    • Alpha cardiac actin antibody
    • alpha sma antibody
    • Alpha-actin-2 antibody
    • Cell growth inhibiting gene 46 protein antibody
    • Cell growth-inhibiting gene 46 protein antibody
    • GIG46 antibody
    • Growth inhibiting gene 46 antibody
    • MYMY5 antibody
    see all

Images

  • Immunocytochemistry/ Immunofluorescence - Anti-alpha smooth muscle Actin antibody [SP171] (ab150301)
    Immunocytochemistry/ Immunofluorescence - Anti-alpha smooth muscle Actin antibody [SP171] (ab150301)
    ab150301 staining alpha smooth muscle Actin in SV40LT-SMC cells (positive control, top panel) and A431 cells (negative control, bottom panel). The cells were fixed with 100% methanol (5 min) then permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab150301 at 1μg/ml concentration and ab7291 (Mouse monoclonal to alpha Tubulin) at 1/1000 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor® 594) (ab150120) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
    Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody [SP171] (ab150301)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody [SP171] (ab150301)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Rat colon tissue sections labeling alpha smooth muscle Actin with ab150301 at 1/200 dilution (0.26 µg/ml). Heat mediated antigen retrieval was performed Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 10 mins. Goat Anti-Rabbit & Mouse IgG (HRP) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
  • Immunocytochemistry/ Immunofluorescence - Anti-alpha smooth muscle Actin antibody [SP171] (ab150301)
    Immunocytochemistry/ Immunofluorescence - Anti-alpha smooth muscle Actin antibody [SP171] (ab150301)
    ab150301 staining alpha smooth muscle Actin in wild-type HeLa cells (top panel) and ACTA2 knockout HeLa cells (bottom panel). The cells were fixed with 100% methanol (5 min) then permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab150301 at 5μg/ml concentration and ab7291 (Mouse monoclonal to alpha Tubulin) at 1/1000 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor® 594) (ab150120) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
    Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).
  • Western blot - Anti-alpha smooth muscle Actin antibody [SP171] (ab150301)
    Western blot - Anti-alpha smooth muscle Actin antibody [SP171] (ab150301)
    All lanes : Anti-alpha smooth muscle Actin antibody [SP171] (ab150301) at 1/130 dilution

    Lane 1 : Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 2 : ACTA2 knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 42 kDa
    Observed band size: 42 kDa



    Lanes 1 - 2: Merged signal (red and green). Green - ab150301 observed at 42 kDa. Red - loading control, ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.

    ab150301 was shown to react with alpha smooth muscle Actin in wild-type HeLa cells in western blot. Loss of signal was observed when ACTA2 knockout sample was used. Wild-type HeLa and ACTA2 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab150301 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4°C at a 1 in 130 Dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody [SP171] (ab150301)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody [SP171] (ab150301)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Mouse colon tissue sections labeling alpha smooth muscle Actin with ab150301 at 1/200 dilution (0.26 µg/ml). Heat mediated antigen retrieval was performed Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 10 mins. Goat Anti-Rabbit & Mouse IgG (HRP) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody [SP171] (ab150301)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody [SP171] (ab150301)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human colon tissue sections labeling alpha smooth muscle Actin with ab150301 at 1/200 dilution (0.26 µg/ml). Heat mediated antigen retrieval was performed Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 10 mins. Goat Anti-Rabbit & Mouse IgG (HRP) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody [SP171] (ab150301)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody [SP171] (ab150301)
    Immunohistochemical analysis of formalin fixed, paraffin embedded Human colon tissue labelling alpha smooth muscle Actin with ab150301 at 1/200 dilution.
  • Immunocytochemistry/ Immunofluorescence - Anti-alpha smooth muscle Actin antibody [SP171] (ab150301)
    Immunocytochemistry/ Immunofluorescence - Anti-alpha smooth muscle Actin antibody [SP171] (ab150301)

    Immunocytochemistry/ Immunofluorescence analysis of NIH/3T3 (mouse embryonic fibroblast) cells labeling alpha smooth muscle Actin with purified ab150301. Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

  • Western blot - Anti-alpha smooth muscle Actin antibody [SP171] (ab150301)
    Western blot - Anti-alpha smooth muscle Actin antibody [SP171] (ab150301)
    Anti-alpha smooth muscle Actin antibody [SP171] (ab150301) at 1/50 dilution + HeLa cell lysate

    Predicted band size: 42 kDa

  • Flow Cytometry (Intracellular) - Anti-alpha smooth muscle Actin antibody [SP171] (ab150301)
    Flow Cytometry (Intracellular) - Anti-alpha smooth muscle Actin antibody [SP171] (ab150301)

    Flow Cytometry analysis of HeLa (human cervix adenocarcinoma epithelial cell) cells labeling alpha smooth muscle Actin with purified ab150301 (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody. Isotype control - Rabbit monoclonal IgG (ab172730) / Black. Unlabeled control - Unlabelled cells / blue.

  • Flow Cytometry (Intracellular) - Anti-alpha smooth muscle Actin antibody [SP171] (ab150301)
    Flow Cytometry (Intracellular) - Anti-alpha smooth muscle Actin antibody [SP171] (ab150301)

    Flow Cytometry analysis of C6 (Rat glial tumor glial cell) cells labeling alpha smooth muscle Actin with purified ab150301 (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used. Isotype control - Rabbit monoclonal IgG (ab172730) / Black. Unlabeled control - Unlabelled cells / blue.

  • Flow Cytometry - Anti-alpha smooth muscle Actin antibody [SP171] (ab150301)
    Flow Cytometry - Anti-alpha smooth muscle Actin antibody [SP171] (ab150301)

    Flow Cytometry analysis of NIH/3T3 (Mouse embryonic fibroblast) cells labeling alpha smooth muscle Actin with purified ab150301 at 1/200 dilution (0.825 µg/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (ab172730) / Black. Unlabeled control - Unlabelled cells / blue.

  • Anti-alpha smooth muscle Actin antibody [SP171] (ab150301)
    Anti-alpha smooth muscle Actin antibody [SP171] (ab150301)

Protocols

  • Western blot protocols
  • Immunohistochemistry protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (6)

Publishing research using ab150301? Please let us know so that we can cite the reference in this datasheet.

ab150301 has been referenced in 6 publications.

  • Zhou Y  et al. The Role of miR-31-5p in the Development of Intervertebral Disc Degeneration and Its Therapeutic Potential. Front Cell Dev Biol 9:633974 (2021). PubMed: 33816484
  • Marcher AB  et al. Transcriptional regulation of Hepatic Stellate Cell activation in NASH. Sci Rep 9:2324 (2019). PubMed: 30787418
  • Keren L  et al. MIBI-TOF: A multiplexed imaging platform relates cellular phenotypes and tissue structure. Sci Adv 5:eaax5851 (2019). Mass Cytometry . PubMed: 31633026
  • Keren L  et al. A Structured Tumor-Immune Microenvironment in Triple Negative Breast Cancer Revealed by Multiplexed Ion Beam Imaging. Cell 174:1373-1387.e19 (2018). Mass Cytometry . PubMed: 30193111
  • Chu H  et al. Effects of transplanted adipose derived stem cells on the expressions of a-SMA and DCN in fibroblasts of hypertrophic scar tissues in rabbit ears. Exp Ther Med 16:1729-1734 (2018). PubMed: 30186394
  • Wang X  et al. MicroRNA-155 targets myosin light chain kinase to inhibit the migration of human bone marrow-derived mesenchymal stem cells. Int J Mol Med 42:1585-1592 (2018). PubMed: 29901087

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