Recombinant Anti-alpha smooth muscle Actin (acetyl E3) + ACTG2 (acetyl E3) antibody [E184] (ab32575)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [E184] to alpha smooth muscle Actin (acetyl E3) + ACTG2 (acetyl E3)
- Suitable for: ELISA, Flow Cyt (Intra), IHC-Fr, WB, IHC-P, ICC/IF
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-alpha smooth muscle Actin (acetyl E3) + ACTG2 (acetyl E3) antibody [E184] -
Description
Rabbit monoclonal [E184] to alpha smooth muscle Actin (acetyl E3) + ACTG2 (acetyl E3) -
Host species
Rabbit -
Specificity
For immunohistochemistry, this antibody only detects actin in smooth muscle and not cardiac muscle.
The antibody has been shown to detect P62736-ACTA (gene ACTA2): (Ac)-EEEDSTALVC and P63267-ACTH (gene ACTG2): (Ac)-EEETTALVC in indirect ELISA.
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Tested applications
Suitable for: ELISA, Flow Cyt (Intra), IHC-Fr, WB, IHC-P, ICC/IFmore details
Unsuitable for: IP -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: A431, HeLa, C6, RAW264.7, PC-12, NIH/3T3 and MCF7 cell lysates. IHC-P: Human uterus, human smooth muscle and mouse smooth muscle tissues. Flow Cyt (intra): HeLa cells. ICC/IF: A431, C6, NIH/3T3 cells.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
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.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
E184 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
- Alexa Fluor® 647 Anti-alpha smooth muscle Actin (acetyl E3) + ACTG2 (acetyl E3) antibody [E184] (ab196919)
- HRP Anti-alpha smooth muscle Actin (acetyl E3) + ACTG2 (acetyl E3) antibody [E184] (ab196920)
- PE Anti-alpha smooth muscle Actin (acetyl E3) + ACTG2 (acetyl E3) antibody [E184] (ab209435)
- Alexa Fluor® 405 Anti-alpha smooth muscle Actin (acetyl E3) + ACTG2 (acetyl E3) antibody [E184] (ab210128)
- Anti-alpha smooth muscle Actin (acetyl E3) + ACTG2 (acetyl E3) antibody [E184] - BSA and Azide free (ab215368)
- FITC Anti-alpha smooth muscle Actin (acetyl E3) + ACTG2 (acetyl E3) antibody [E184] (ab223920)
- APC Anti-alpha smooth muscle Actin (acetyl E3) + ACTG2 (acetyl E3) antibody [E184] (ab223921)
- Anti-Vimentin antibody [LN-6] (ab230171)
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
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.
Application | Abreviews | Notes |
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ELISA |
Use a concentration of 2e-005 - 1 µg/ml.
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Flow Cyt (Intra) |
1/20.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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IHC-Fr | (4) |
Use at an assay dependent concentration.
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WB | (3) |
1/1000 - 1/5000. Detects a band of approximately 42 kDa (predicted molecular weight: 42 kDa).
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IHC-P | (10) |
1/200 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
See IHC antigen retrieval protocols. Internal test showed non-specific staining on mouse kidney, mouse stomach, rat kidney and rat stomach. |
ICC/IF | (3) |
1/500.
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Notes |
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ELISA
Use a concentration of 2e-005 - 1 µg/ml. |
Flow Cyt (Intra)
1/20. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
IHC-Fr
Use at an assay dependent concentration. |
WB
1/1000 - 1/5000. Detects a band of approximately 42 kDa (predicted molecular weight: 42 kDa). |
IHC-P
1/200 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. See IHC antigen retrieval protocols. Internal test showed non-specific staining on mouse kidney, mouse stomach, rat kidney and rat stomach. |
ICC/IF
1/500. |
Target
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Cellular localization
alpha smooth muscle Actin: Cytoplasm > cytoskeleton. ACTG2: Cytoplasm > cytoskeleton. -
Database links
- Entrez Gene: 59 Human
- Entrez Gene: 72 Human
- Entrez Gene: 11468 Mouse
- Entrez Gene: 11475 Mouse
- Entrez Gene: 25365 Rat
- Entrez Gene: 81633 Rat
- Omim: 102545 Human
- Omim: 102620 Human
see all
Images
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin (acetyl E3) + ACTG2 (acetyl E3) antibody [E184] (ab32575)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human smooth muscle tissue labeling 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, an 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.
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Immunocytochemistry/ Immunofluorescence - Anti-alpha smooth muscle Actin (acetyl E3) + ACTG2 (acetyl E3) antibody [E184] (ab32575)
Immunocytochemistry/ Immunofluorescence analysis of NIH/3T3(Mouse embryonic fibroblast) cells labeling alpha smooth muscle Actin with purified ab32575 at 1/500 dilution (5.2 µg/ml). Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 dilution (2 µg/ml) dilution. DAPI was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
Alexa Fluor® 488 (ab197240) and Alexa Fluor® 647 (ab196919) conjugated versions are available for this clone.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin (acetyl E3) + ACTG2 (acetyl E3) antibody [E184] (ab32575)Bjerke GA. et al PLoS One. 2014 Mar 20;9(3):e92800. doi: 10.1371/journal.pone.0092800. eCollection 2014. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/
FoxA1 (red) and alpha smooth muscle actin (green) staining are shown by indirect immunofluorescence on sections of prostate from mice of the indicated genotypes.
Wild type: 21 weeks, Tgfbr2r/r: 44 weeks, Ptenr/r: 21 weeks, Ptenr/r;Tgfbr2r/r: 11 weeks, Apcr/r: 36 weeks, and Apcr/r;Tgfbr2r/r: 24 weeks old.
IF images were captured on an Olympus BX51 microscope and DP70 digital camera, or on a Nikon Eclipse NI-U and captured with a DS-QI1 camera with NIS Elements software.
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Western blot - Anti-alpha smooth muscle Actin (acetyl E3) + ACTG2 (acetyl E3) antibody [E184] (ab32575)All lanes : Anti-alpha smooth muscle Actin (acetyl E3) + ACTG2 (acetyl E3) antibody [E184] (ab32575) at 1/5000 dilution (purified)
Lane 1 : C6 (Rat glial tumor cell line) whole cell lysate
Lane 2 : RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate
Lane 3 : PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate
Lane 4 : NIH/3T3 (Mouse embryo fibroblast cell line) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 42 kDa
Observed band size: 42 kDaBlocking and dilution buffer: 5% NFDM/TBST
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Flow Cytometry (Intracellular) - Anti-alpha smooth muscle Actin (acetyl E3) + ACTG2 (acetyl E3) antibody [E184] (ab32575)
Intracellular Flow Cytometry analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling 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 - Unlabeled control, cells without incubation with primary and secondary antibodies.
Alexa Fluorr®488 (ab197240) and Alexa Fluorr®647 (ab196919) conjugated versions are available for this clone.
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Immunocytochemistry/ Immunofluorescence - Anti-alpha smooth muscle Actin (acetyl E3) + ACTG2 (acetyl E3) antibody [E184] (ab32575)Immunocytochemistry/ Immunofluorescence analysis of C6(Rat glial tumor glial cell) cells labeling alpha smooth muscle Actin with purified ab32575 at 1/100 dilution (0.71 µg/ml). Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 dilution (2 µg/ml) dilution. DAPI was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
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Western blot - Anti-alpha smooth muscle Actin (acetyl E3) + ACTG2 (acetyl E3) antibody [E184] (ab32575)Different batches of ab32575 were tested on A431 (Human epidermoid carcinoma epithelial cell) lysate at 0.005 µg/ml. 15 µg of lysate was loaded in each lane. Bands observed at 42 kDa.
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Western blot - Anti-alpha smooth muscle Actin (acetyl E3) + ACTG2 (acetyl E3) antibody [E184] (ab32575)All lanes : Anti-alpha smooth muscle Actin (acetyl E3) + ACTG2 (acetyl E3) antibody [E184] (ab32575) at 1/5000 dilution (purified)
Lane 1 : A431 (Human epidermoid carcinoma cell line) whole cell lysate
Lane 2 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 3 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 42 kDa
Observed band size: 42 kDaBlocking and dilution buffer: 5% NFDM/TBST
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Immunocytochemistry/ Immunofluorescence - Anti-alpha smooth muscle Actin (acetyl E3) + ACTG2 (acetyl E3) antibody [E184] (ab32575)
Immunocytochemistry/Immunofluorescence analysis of A431 (human epidermoid carcinoma) cells labeling 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 counterstained with ab7291, anti-Tubulin (mouse mAb) at 1/1000 followed by ab150120 Alexa Fluor®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).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin (acetyl E3) + ACTG2 (acetyl E3) antibody [E184] (ab32575)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse smooth muscle tissue labeling 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, an 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.
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Following reconstitution in PBS with 10% DMSO, peptides (1 ug per mL) were immobilised in PBS on an ELISA plate overnight. After blocking in 5% BSA, primary antibody (ab271180) was added in a concentration range of 0.017-1000 ng per mL.
Pre-adsorbed secondary antibody goat anti-rabbit IgG H&L (HRP, ab97080) was used at 1/20000 dilution.
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Following reconstitution in PBS with 10% DMSO, peptides (1 ug per mL) were immobilised in PBS on an ELISA plate overnight. After blocking in 5% BSA, primary antibody (ab271180) was added in a concentration range of 0.017-1000 ng per mL.
Pre-adsorbed secondary antibody goat anti-rabbit IgG H&L (HRP, ab97080) was used at 1/20000 dilution.
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.
Datasheets and documents
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SDS download
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Datasheet download
References (532)
ab32575 has been referenced in 532 publications.
- Wu Q et al. Twist1 regulates macrophage plasticity to promote renal fibrosis through galectin-3. Cell Mol Life Sci 79:137 (2022). PubMed: 35182235
- Zheng S et al. circCUL2 induces an inflammatory CAF phenotype in pancreatic ductal adenocarcinoma via the activation of the MyD88-dependent NF-κB signaling pathway. J Exp Clin Cancer Res 41:71 (2022). PubMed: 35189958
- Zha Y et al. ADAMTS8 Promotes Cardiac Fibrosis Partly Through Activating EGFR Dependent Pathway. Front Cardiovasc Med 9:797137 (2022). PubMed: 35224040
- Wang S et al. Obstructive Sleep Apnea Affects Lacrimal Gland Function. Invest Ophthalmol Vis Sci 63:3 (2022). PubMed: 35238868
- Wang PW et al. Irisin alleviates vascular calcification by inhibiting VSMC osteoblastic transformation and mitochondria dysfunction via AMPK/Drp1 signaling pathway in chronic kidney disease. Atherosclerosis 346:36-45 (2022). PubMed: 35255258