Overview

  • Product name
    Anti-alpha smooth muscle Actin antibody [1A4] (FITC)
    See all alpha smooth muscle Actin primary antibodies
  • Description
    Mouse monoclonal [1A4] to alpha smooth muscle Actin (FITC)
  • Host species
    Mouse
  • Conjugation
    FITC. Ex: 493nm, Em: 528nm
  • Tested applications
    Suitable for: ICC/IF, Flow Cyt, IHC-FoFr, IHC-P, IHC-Fr, WBmore details
  • Species reactivity
    Reacts with: Rat, Human
    Predicted to work with: Mouse, Sheep, Goat, Chicken, Guinea pig, Cat, Dog, Snake, Zebrafish
  • Immunogen

    The N-terminal decapeptide of alpha-smooth muscle actin.

  • Positive control
    • In IF, this antibody gave a positive signal in SV40LT-SMC cells.
  • 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.

Properties

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.
Flow Cyt Use at an assay dependent concentration. PubMed: 17222057

ab18455 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.

IHC-FoFr Use at an assay dependent concentration.
IHC-P 1/50 - 1/75. (as well as on cell smears).
IHC-Fr Use at an assay dependent concentration.
WB Use at an assay dependent concentration. PubMed: 17237513Use at an assay dependent dilution (from Pubmed:17237513).

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
  • 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

  • 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).

References

This product has been referenced in:
  • 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 . Read more (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 . Read more (PubMed: 29216542) »
See all 35 Publications for this product

Customer reviews and Q&As

1-10 of 10 Abreviews or Q&A

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
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

Ms. Chelsea Stowell

Verified customer

Submitted Mar 07 2018

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

Answer

Thank you for contacting us. Unfortunately we do not have a permeabilizing kit for flow cytometry in our catalogue for the moment. This antibody should however work with the general classical permeabilization methods. If there should be a problem, please provide us with the details and we will investigate the case as usual. There are several companies offering commercial permeabilization reagents for flow cytometry. We do also have an online protocol for intracellular flow cytometry staining. Please feel free to forward the link to the customer. https://www.abcam.com/index.html?pageconfig=resource&rid=11448 I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Read More
Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (Aortic smooth muscle cells)
Specification
Aortic smooth muscle cells
Fixative
Paraformaldehyde
Permeabilization
Yes - Triton-X/methanol
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C

Abcam user community

Verified customer

Submitted May 29 2008

Question
Answer

Thank you for your enquiry. The isotype listed on the data sheet is IgG2A. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

Read More

Answer

Thank you for your enquiry. Unfortunately this antiserum, alpha smooth muscle Actin ab8211 does not contain an IgG2a kappa isotype control. This antiserum is IgG sera. We do stock a mouse IgG2a, kappa isotype control (ab18454). I hope this information helps, please do not hesitate to contact me should you require further assistance.

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Answer

Dear Angela As actin is highly conserved, I would expect ab8211 to work on rat tissue. I would be willing to say this with a strong degree of confidence. I am afraid that we don't have a polyclonal to alpha actin at the moment. I hope this helps.

Read More

Answer

Dear Susan This antibody should react with mouse a-smooth muscle actin. I suggest starting with a range of dilutions at 1:50, 1:100 and 1:500. This should give you a good idea of how to optimise it. Obviously the dilution required depends on a number of factors. I hope this helps. Regards Michael

Read More

Answer

This antibody has not yet been tested for cross reactivity with chicken.

Read More

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