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Validated using a knockout cell lineRecombinant

Recombinant Anti-alpha smooth muscle Actin antibody [1A4] - BSA and Azide free (ab240654)

Reviews (1) Submit a question References (1)
Immunocytochemistry/ Immunofluorescence - Anti-alpha smooth muscle Actin antibody [1A4] - BSA and Azide free (ab240654)
  • Immunocytochemistry/ Immunofluorescence - Anti-alpha smooth muscle Actin antibody [1A4] - BSA and Azide free (ab240654)
  • Immunocytochemistry/ Immunofluorescence - Anti-alpha smooth muscle Actin antibody [1A4] - BSA and Azide free (ab240654)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody [1A4] - BSA and Azide free (ab240654)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody [1A4] - BSA and Azide free (ab240654)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody [1A4] - BSA and Azide free (ab240654)
  • Western blot - Anti-alpha smooth muscle Actin antibody [1A4] - BSA and Azide free (ab240654)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Mouse monoclonal [1A4] to alpha smooth muscle Actin - BSA and Azide free
  • Suitable for: WB, IHC-P, ICC/IF
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 594 Biotin FITC HRP

Overview

  • Product name

    Anti-alpha smooth muscle Actin antibody [1A4] - BSA and Azide free
    See all alpha smooth muscle Actin primary antibodies

  • Description

    Mouse monoclonal [1A4] to alpha smooth muscle Actin - BSA and Azide free

  • Host species

    Mouse

  • Tested applications

    Suitable for: WB, IHC-P, ICC/IFmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • ICC/IF: NIH-3T3 (mouse embryonic fibroblast) cells IHC: Human colon, Mouse colon, Rat colon WB: NIH-3T3 (mouse embryonic fibroblast) whole cell lysate, IMR-90 (human lung fibroblast) whole cell lysate

  • General notes

    This product has switched to a recombinant production method on 15th September 2021.

    ab240654 is the carrier-free version of ab7817.

    This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab240654 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
WB
1/1000. Detects a band of approximately 42 kDa (predicted molecular weight: 42 kDa).
IHC-P (1)
1/50000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF
1/50.

Not recommended for Human.
Notes
WB
1/1000. Detects a band of approximately 42 kDa (predicted molecular weight: 42 kDa).
IHC-P
1/50000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF
1/50.

Not recommended for Human.

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.
  • Information by UniProt

  • Database links

    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 [1A4] - BSA and Azide free (ab240654)
    Immunocytochemistry/ Immunofluorescence - Anti-alpha smooth muscle Actin antibody [1A4] - BSA and Azide free (ab240654)
    This data was developed using the same antibody clone in a different buffer formulation (ab7817). ab7817 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 ab7817 at 1μg/ml concentration and ab6046 (Rabbit polyclonal to beta 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 mouse IgG (Alexa Fluor® 488) (ab150117) at 2 μg/ml (shown in green) and a goat secondary antibody to rabbit IgG (Alexa Fluor® 594) (ab150080) 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).
  • Immunocytochemistry/ Immunofluorescence - Anti-alpha smooth muscle Actin antibody [1A4] - BSA and Azide free (ab240654)
    Immunocytochemistry/ Immunofluorescence - Anti-alpha smooth muscle Actin antibody [1A4] - BSA and Azide free (ab240654)

    This data was developed using ab7817, the same antibody clone in a different buffer formulation.

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NIH 3T3 cells labelling alpha smooth muscle Actin with ab7817 at 1/50 dilution (Green). Confocal image showing cytoskeleton staining in NIH 3T3 cells. ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed was used as a secondary at 1/10 dilution. ab179513 Anti-beta Tubulin rabbit monoclonal antibody was used to counterstain tubulin at 1/20 dilution, followed by ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) at 1/50 dilution (Red). The Nuclear counterstain was DAPI (Blue).


    Negative control: secondary counterstain antibody only control. 

     

     

     

  • Immunocytochemistry/ Immunofluorescence - Anti-alpha smooth muscle Actin antibody [1A4] - BSA and Azide free (ab240654)
    Immunocytochemistry/ Immunofluorescence - Anti-alpha smooth muscle Actin antibody [1A4] - BSA and Azide free (ab240654)

    This data was developed using ab7817, the same antibody clone in a different buffer formulation.

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NIH 3T3 cells labelling alpha smooth muscle Actin with ab7817 at 1/50 dilution (Green). Confocal image showing cytoskeleton staining in NIH 3T3 cells. ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed was used as a secondary at 1/10 dilution. ab179513 Anti-beta Tubulin rabbit monoclonal antibody was used to counterstain tubulin at 1/20 dilution, followed by ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) at 1/50 dilution (Red). The Nuclear counterstain was DAPI (Blue).


    Negative control: secondary counterstain antibody only control. 

     

     

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody [1A4] - BSA and Azide free (ab240654)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody [1A4] - BSA and Azide free (ab240654)

    Immunohistochemical analysis of Rat colon tissue labelling alpha smooth muscle Actin with ab7817 at 1/50000 dilution and ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) Secondary antibody. Positive cytoplasmic staining on Rat colon tissue is observed. Counterstained with Hematoxylin. 

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody was ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). 

    This image is produced using the same clone but in a different formulation ab7817.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody [1A4] - BSA and Azide free (ab240654)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody [1A4] - BSA and Azide free (ab240654)

    Immunohistochemical analysis of Mouse colon tissue labelling alpha smooth muscle Actin with ab7817 at 1/50000 dilution and ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) Secondary antibody. Positive cytoplasmic staining on Mouse colon tissue is observed. Counterstained with Hematoxylin. 

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody was ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). 

    This image is produced using the same clone but in a different formulation ab7817.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody [1A4] - BSA and Azide free (ab240654)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody [1A4] - BSA and Azide free (ab240654)

    Immunohistochemical analysis of Human colon tissue labelling alpha smooth muscle Actin with ab7817 at 1/50000 dilution and ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) Secondary antibody. Positive cytoplasmic staining on Human colon tissue is observed. Counterstained with Hematoxylin. 

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody was ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). 

    This image is produced using the same clone but in a different formulation ab7817.

  • Western blot - Anti-alpha smooth muscle Actin antibody [1A4] - BSA and Azide free (ab240654)
    Western blot - Anti-alpha smooth muscle Actin antibody [1A4] - BSA and Azide free (ab240654)
    All lanes : Anti-alpha smooth muscle Actin antibody [1A4] (ab7817) at 1/1000 dilution

    Lane 1 : NIH 3T3 (mouse embryonic fibroblast) whole cell lysate with NFDM/TBST
    Lane 2 : IMR-90 (human lung fibroblast), whole cell lysate with NFDM/TBST

    Lysates/proteins at 20 µg per lane.

    Blocking peptides at 5 % per lane.

    Secondary
    All lanes : Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution

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



    This image is produced using the same clone but in a different formulation ab7817.

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

References (1)

Publishing research using ab240654? Please let us know so that we can cite the reference in this datasheet.

ab240654 has been referenced in 1 publication.

  • Choi B  et al. Evogliptin Suppresses Calcific Aortic Valve Disease by Attenuating Inflammation, Fibrosis, and Calcification. Cells 10:N/A (2021). PubMed: 33401457

Customer reviews and Q&As

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-alpha smooth muscle Actin antibody [1A4] - BSA and Azide free

 Excellent
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Colorectal Carcinoma (CRC))
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: pH6 IHC Antigen Retrieval Solution (Invitrogen low pH 00-4955-58)
Permeabilization
No
Specification
Colorectal Carcinoma (CRC)
Blocking step
protein block - serum free 0.25% casein in PBS, containing stabilizing protein and 0.015 mol/L sodium azide as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 0.25% · Temperature: 21°C
Fixative
Formaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Miss. Amy Burley

Verified customer

Submitted Dec 22 2022

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