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  1. Link

    fitc-alpha-smooth-muscle-actin-antibody-1a4-ab8211.pdf

  1. Send me a copy of this email
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Signal Transduction Cytoskeleton / ECM Cytoskeleton Microfilaments Actin etc Actin
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Validated using a knockout cell line

FITC Anti-alpha smooth muscle Actin antibody [1A4] (ab8211)

  • Datasheet
  • SDS
Reviews (9)Q&A (8)References (41)

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Western blot - FITC Anti-alpha smooth muscle Actin antibody [1A4] (ab8211)
  • Immunocytochemistry - FITC Anti-alpha smooth muscle Actin antibody [1A4] (ab8211)
  • Immunocytochemistry/ Immunofluorescence - FITC Anti-alpha smooth muscle Actin antibody [1A4] (ab8211)

Key features and details

  • FITC Mouse monoclonal [1A4] to alpha smooth muscle Actin
  • Suitable for: ICC/IF, WB
  • Knockout validated
  • Reacts with: Rat
  • Conjugation: FITC. Ex: 493nm, Em: 528nm
  • Isotype: IgG2a

Overview

  • Product name

    FITC Anti-alpha smooth muscle Actin antibody [1A4]
    See all alpha smooth muscle Actin primary antibodies
  • Description

    FITC Mouse monoclonal [1A4] to alpha smooth muscle Actin
  • Host species

    Mouse
  • Conjugation

    FITC. Ex: 493nm, Em: 528nm
  • Tested applications

    Suitable for: ICC/IF, WBmore details
  • Species reactivity

    Reacts with: Rat
    Predicted to work with: Mouse, Sheep, Goat, Chicken, Guinea pig, Cat, Dog, Human, Snake, Zebrafish
  • Immunogen

    corresponding to alpha smooth muscle Actin.

  • Positive control

    • ICC: HeLa and SV40LT-SMC cells. WB: HeLa cell lysate.
  • General notes

    This antibody reacts with smooth muscle cells of blood vessels and parenchymal tissue of intestine, testis and ovary. This antibody strongly crossreacts with alphasmooth muscle actin in a variety of mammalian and non-mammalian species.

    The antibody is conjugated with FITC at a ratio 6 Moles of Fluorescein to 1 Mole of antibody. Maximum excitation of FITC occurs at wavelength 492 nm, and maximum emission occurs at wavelenght 520 nm.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    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, as well as customer reviews and Q&As.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C. Avoid freeze / thaw cycle. Store In the Dark.
  • Storage buffer

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituents: PBS, 50% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Primary antibody notes

    This antibody reacts with smooth muscle cells of blood vessels and parenchymal tissue of intestine, testis and ovary. This antibody strongly crossreacts with alphasmooth muscle actin in a variety of mammalian and non-mammalian species.
  • Clonality

    Monoclonal
  • Clone number

    1A4
  • Isotype

    IgG2a
  • Light chain type

    kappa
  • 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

    • Biotin Anti-alpha smooth muscle Actin antibody [1A4] (ab125057)
    • Alexa Fluor® 488 Anti-alpha smooth muscle Actin antibody [1A4] (ab184675)
    • Alexa Fluor® 594 Anti-alpha smooth muscle Actin antibody [1A4] (ab202368)
    • HRP Anti-alpha smooth muscle Actin antibody [1A4] (ab203696)
    • Anti-alpha smooth muscle Actin antibody [1A4] - BSA and Azide free (ab240654)
    • Anti-alpha smooth muscle Actin antibody [1A4] (ab7817)
  • Isotype control

    • FITC Mouse IgG2a, kappa monoclonal [MOPC-173] - isotype control (ab18455)
  • Recombinant Protein

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

    • Phalloidin-iFluor 488 Reagent (ab176753)
    • Phalloidin-iFluor 647 Reagent (ab176759)
    • Alexa Fluor® 647 Anti-Tubulin antibody [YOL1/34] - Microtubule Marker (ab195884)

Applications

Our Abpromise guarantee covers the use of ab8211 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
ICC/IF Use a concentration of 5 µg/ml.
WB 1/131.58. Predicted molecular weight: 42 kDa.

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: 101093547 Cat
    • Entrez Gene: 423787 Chicken
    • Entrez Gene: 477587 Dog
    • Entrez Gene: 59 Human
    • Entrez Gene: 11475 Mouse
    • Entrez Gene: 81633 Rat
    • Entrez Gene: 101121051 Sheep
    • Entrez Gene: 322509 Zebrafish
    • Omim: 102620 Human
    • SwissProt: P08023 Chicken
    • SwissProt: P62736 Human
    • SwissProt: P62737 Mouse
    • 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

  • Western blot - FITC Anti-alpha smooth muscle Actin antibody [1A4] (ab8211)
    Western blot - FITC Anti-alpha smooth muscle Actin antibody [1A4] (ab8211)
    All lanes : Anti-alpha smooth muscle Actin antibody [1A4] (ab7817) at 1/131.58 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : ACTA2 knockout HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

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



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

    Lanes 1 - 2: Merged signal (red and green). Green - ab7817 observed at 42 kDa. Red - loading control, ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) observed at 55kDa.

    ab7817 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 ab7817 and ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) overnight at 4°C at a 1 in 131.58 Dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • Immunocytochemistry - FITC Anti-alpha smooth muscle Actin antibody [1A4] (ab8211)
    Immunocytochemistry - FITC Anti-alpha smooth muscle Actin antibody [1A4] (ab8211)
    ab8211 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 ab8211 at 5μg/ml concentration and ab195884 (Rat monoclonal to Tubulin - Alexa Fluor® 647) at 1/100 dilution overnight at 4°C. Nuclear DNA was labelled in blue with DAPI.
    Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).
  • Immunocytochemistry/ Immunofluorescence - FITC Anti-alpha smooth muscle Actin antibody [1A4] (ab8211)
    Immunocytochemistry/ Immunofluorescence - FITC Anti-alpha smooth muscle Actin antibody [1A4] (ab8211)

    ab8211 staining alpha smooth muscle Actin in SV40LT-SMC cells. The cells were fixed with 100% methanol (5 min), 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 overnight at +4°C with ab8211 at a 1/100 dilution (shown in green) and ab195889, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 594), at a 1/250 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

    This product also gave a positive signal under the same testing conditions in SV40LT-SMC cells fixed with 4% formaldehyde (10 min).

Protocols

  • Immunocytochemistry & immunofluorescence protocols

Click here to view the general protocols

Datasheets and documents

    • Datasheet
    • SDS
  • References (41)

    Publishing research using ab8211? Please let us know so that we can cite the reference in this datasheet.

    ab8211 has been referenced in 41 publications.

    • Yu J  et al. PLK1 Inhibition alleviates transplant-associated obliterative bronchiolitis by suppressing myofibroblast differentiation. Aging (Albany NY) 12:11636-11652 (2020). PubMed: 32541091
    • El-Kehdy H  et al. Inflammation Differentially Modulates the Biological Features of Adult Derived Human Liver Stem/Progenitor Cells. Cells 9:N/A (2020). PubMed: 32650454
    • Bogdanova M  et al. Interstitial cells in calcified aortic valves have reduced differentiation potential and stem cell-like properties. Sci Rep 9:12934 (2019). PubMed: 31506459
    • Zhou X  et al. CDKN1A-interacting zinc finger protein 1 is a novel biomarker for lung squamous cell carcinoma. Oncol Lett 15:183-188 (2018). IF ; Human . PubMed: 29285191
    • Lai YL  et al. Loss of heme oxygenase-1 accelerates mesodermal gene expressions during embryoid body development from mouse embryonic stem cells. Redox Biol 15:51-61 (2018). Flow Cyt . PubMed: 29216542
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-10 of 17 Abreviews or Q&A

    Question

     Would you be able to provide our customer with a protocol for Flow Cytometry staining (which is best suited for your Smooth Muscle Actin Antibody, AB8211)? There are a number of protocols on the suggested website (https://www.abcam.com/index.html?pageconfig=resource&rid=11448) and would appreciate if you are able to advise which would best work with this antibody, AB8211.   With many thanks and best regards,,

    Read More

    Abcam community

    Verified customer

    Asked on Sep 23 2011

    Answer

    Thank you very much for your question. Indeed, the use of ab8211 in flow cytometry has been documented in the publication "Delayed Goblet Cell Hyperplasia, Acetylcholine Receptor Expression, and Worm Expulsion in SMC-Specific IL-4Rα–Deficient Mice " PMID 17222057 http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1769405/?tool=pubmed "For the intracellular stain, cells in single cell suspension were incubated with 1% normal rat serum and stained with rat anti-mouse IL-4Rα–PE (mIL-4RM-1; BD Biosciences, http://www.bdbiosciences.com). Stained cells were then washed, fixed in 2% paraformaldehyde, permeabilized with saponin, preblocked with 2% NRS and Fc block (2.4G2), and stained with anti–α-actin FITC (Abcam, https://www.abcam.com) or isotype control IgG2a-FITC (BD Biosciences). " I hope this helps the customer. In conjunction with the protocol on intracellular flow cytometry https://www.abcam.com/index.html?pageconfig=resource&rid=11448, this should allow the reproduction of good results. Please let me know if the customer needs more information. Please let me also know if the customer has any problems. I would be pleased to investigate this for him. As you know, we do cover all our products to work as stated on the datasheet. I wish you a great week.

    Read More

    Abcam Scientific Support

    Answered on Sep 23 2011

    Immunohistochemistry (PFA perfusion fixed frozen sections) abreview for Anti-alpha smooth muscle Actin antibody [1A4] (FITC)

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (PFA perfusion fixed frozen sections)
    Sample
    Rat Tissue sections (Kidney)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citrate Buffer pH 6.0
    Permeabilization
    Yes - 0.05% Tween 20
    Specification
    Kidney
    Blocking step
    Sea Block as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 22°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Feb 26 2020

    Immunocytochemistry/ Immunofluorescence abreview for Anti-alpha smooth muscle Actin antibody [1A4] (FITC)

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (Kidney H9 Organoid)
    Permeabilization
    Yes - 0.05% Tween 20
    Specification
    Kidney H9 Organoid
    Blocking step
    Sea Block as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 22°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Feb 08 2020

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

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Pig Tissue sections (Kidney)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citrate Buffer pH 6.0
    Permeabilization
    Yes - 0.05% Tween 20
    Specification
    Kidney
    Blocking step
    Sea Block as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 22°C
    Fixative
    Formaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Feb 08 2020

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

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (Kidney)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: ab64236 100x Citrate Buffer pH 6.0
    Permeabilization
    Yes - 0.05% Tween 20
    Specification
    Kidney
    Blocking step
    Sea Block as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 22°C
    Fixative
    Formaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Jan 15 2020

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

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Dog Tissue sections (Liver)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: ab64236 100x Citrate Buffer pH 6.0
    Permeabilization
    Yes - 0.05% Tween 20
    Specification
    Liver
    Blocking step
    sea block as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 22°C
    Fixative
    Formaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Dec 24 2019

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

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Rat Tissue sections (Kidney)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citrate Buffer pH 6.0
    Permeabilization
    Yes - 0.05% Tween 20
    Specification
    Kidney
    Blocking step
    sea block as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 22°C
    Fixative
    Formaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Dec 24 2019

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

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (Breast)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: ab64236 100x Citrate Buffer pH 6.0
    Permeabilization
    Yes - 0.05% Tween 20
    Specification
    Breast
    Blocking step
    sea block as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 4°C
    Fixative
    Formaldehyde
    Read More

    Mr. David Ivancic

    Verified customer

    Submitted Jul 18 2019

    Immunohistochemistry (Frozen sections) abreview for Anti-alpha smooth muscle Actin antibody [1A4] (FITC)

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Sheep Tissue sections (Artery)
    Permeabilization
    Yes - 0.1% Triton X-100, 10 min
    Specification
    Artery
    Blocking step
    Serum as blocking agent for 4 hour(s) and 30 minute(s) · Concentration: 5% · Temperature: 20°C
    Fixative
    Paraformaldehyde
    Read More

    Dr. Chelsea Stowell

    Verified customer

    Submitted Mar 07 2018

    Question

     Thank you once again for your help extended. Our customer has reverted with the following question regarding the protocol referenced in your earlier email. It was mentioned:“Stained cells were then washed, fixed in 2% paraformaldehyde, permeabilized with saponin, preblocked with 2% NRS and Fc block (2.4G2), and stained with anti–α-actin FITC (Abcam, https://www.abcam.com) or isotype control IgG2a-FITC (BD Biosciences). " Would you be able to advise the purpose of pre-blocking with 2% NRS and Fc block? Is it a necessary step when using Ab8211?   With many thanks and best regards, 

    Read More

    Abcam community

    Verified customer

    Asked on Oct 07 2011

    Answer

    Thank you for your email. Indeed, I can confirm that the protocol is in reference to ab8211. The use of blocking is to decrease unspecific signal and background. The Fc block with the monoclonal antibody 2.4G2 is to block the Fc receptors expressed on certain cells, such as macrophages. The Fc receptors have a high affinity for immunoglobulins and might therefore bind the primary antibody via its Fc chain- resulting in an unspecific staining. The 2.4G2 is directed against Fc receptors and is shown to block their ability to bind immunoglobulins. The additional use of NRS (normal rat serum) is to block unspecific protein-protein interaction. It is therefore dependent on the cell type the customer is analysing whether these two blocking methods are necessary. It is however very good practice to include them and I can recommend to do so. I hope this information is helpful. Please do not hesitate to contact me again should the customer have other questions.

    Read More

    Abcam Scientific Support

    Answered on Oct 07 2011

    1-10 of 17 Abreviews or Q&A

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