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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.
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents
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.
This product is a recombinant rabbit monoclonal antibody.
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|
|IF||Use at an assay dependent concentration. PubMed: 28138565|
|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). Internal test showed non-specific staining on mouse kidney, mouse stomach, rat kidney and rat stomach.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
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.
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 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.
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.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"