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Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human alpha smooth muscle Actin (N terminal). The exact sequence is proprietary.
This product is a recombinant rabbit monoclonal antibody.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated ‘PUR’ on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Alternative versions available:
Anti-alpha smooth muscle Actin antibody (Alexa Fluor® 488) [E184] (ab197240)
Anti-alpha smooth muscle Actin antibody (Alexa Fluor® 647) [E184] (ab196919)
Anti-alpha smooth muscle Actin antibody (HRP) [E184] (ab196920)
Anti-alpha smooth muscle Actin antibody (Phycoerythrin) [E184] (ab209435)
Anti-alpha smooth muscle Actin antibody (Alexa Fluor® 405) [E184] (ab210128)
Produced using Abcam's RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5, 675, 063 and/or 7, 429, 487.
Our Abpromise guarantee covers the use of ab32575 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-FrFl||Use at an assay dependent concentration. PubMed: 24647450|
|WB||1/1000 - 1/5000. Detects a band of approximately 42 kDa (predicted molecular weight: 42 kDa).Can be blocked with Human alpha smooth muscle Actin peptide (ab211918).|
|IHC-P||1/200 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
Immunocytochemistry/Immunofluorescence analysis of A431 (human epidermoid carcinoma) cells labelling alpha smooth muscle Actin (green) with purified ab32575 at 1/500. Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. Cells were counter-stained with ab7291, anti-Tubulin (mouse mAb) at 1/1000 followed by ab150120 AlexaFluor®594 goat anti-mouse secondary (1/1000). Nuclei were counterstained with DAPI (blue).
For negative control 1, rabbit primary antibody and anti-mouse secondary antibody (ab150120) were used. For negative control 2, ab7291 (mouse primary antibody) was used followed by anti-rabbit secondary antibody (ab150077).
Flow Cytometry analysis of HeLa cells labelling alpha smooth muscle Actin with purified ab32575 at a dilution of 1/20 (red). Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human smooth muscle tissue labelling alpha smooth muscle Actin with purified ab32575 at a dilution of 1/200. Heat mediated antigen retrieval was performed using EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse smooth muscle tissue labelling alpha smooth muscle Actin with purified ab32575 at a dilution of 1/200. Heat mediated antigen retrieval was performed using EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
Overlay histogram showing HeLa cells stained with unpurified ab32575 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (unpurified ab32575, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was a goat anti-rabbit DyLight® 488 (IgG; H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
Unpurified ab32575 staining alpha smooth muscle actin in human kidney tumour sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with acetone and blocked with 10% serum for 20 minutes at 21°C. Samples were incubated with primary antibody (1/100 in TBS + 2% BSA + 0.02% sodium azide) for 1 hour at 21°C. An undiluted HRP-conjugated Goat anti-rabbit polyclonal was used as the secondary antibody.