Recombinant Anti-alpha smooth muscle Actin antibody [SP171] - BSA and Azide free (ab242395)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [SP171] to alpha smooth muscle Actin - BSA and Azide free
- Suitable for: ICC/IF, IHC-P, WB, Flow Cyt (Intra), mIHC
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-alpha smooth muscle Actin antibody [SP171] - BSA and Azide free
See all alpha smooth muscle Actin primary antibodies -
Description
Rabbit monoclonal [SP171] to alpha smooth muscle Actin - BSA and Azide free -
Host species
Rabbit -
Tested applications
Suitable for: ICC/IF, IHC-P, WB, Flow Cyt (Intra), mIHCmore details -
Species reactivity
Reacts with: Mouse, Rat, Human
Predicted to work with: Rabbit, Chicken, Cow, Pig -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- IHC-P: Human colon, Mouse colon, and Rat colon tissue. WB: Recombinant Human alpha smooth muscle Actin protein (ab114148); HeLa cell lysate. Flow Cyt (intra): HeLa, NIH/3T3 and C6 cells. ICC: NIH/3T3 and HeLa cells.
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General notes
ab242395 is the carrier-free version of ab150301.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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
This product is FOR RESEARCH USE ONLY. For commercial use, please contact partnerships@abcam.com.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Storage buffer
pH: 7.20
Constituent: PBS -
Carrier free
Yes -
Concentration information loading...
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Purity
Affinity purified -
Purification notes
Purified from TCS by protein A/G. -
Clonality
Monoclonal -
Clone number
SP171 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
- Anti-alpha smooth muscle Actin antibody [SP171] (ab150301)
- Alexa Fluor® 488 Anti-alpha smooth muscle Actin antibody [SP171] (ab267536)
- Alexa Fluor® 647 Anti-alpha smooth muscle Actin antibody [SP171] (ab267537)
- APC Anti-alpha smooth muscle Actin antibody [SP171] (ab310827)
- PE Anti-alpha smooth muscle Actin antibody [SP171] (ab310903)
- Alexa Fluor® 594 Anti-alpha smooth muscle Actin antibody [SP171] (ab311730)
- Alexa Fluor® 568 Anti-alpha smooth muscle Actin antibody [SP171] (ab313007)
- Alexa Fluor® 555 Anti-alpha smooth muscle Actin antibody [SP171] (ab313211)
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Compatible Secondaries
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Conjugation kits
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KO cell lines
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KO cell lysates
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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 ab242395 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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ICC/IF |
Use at an assay dependent concentration.
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IHC-P | (2) |
Use at an assay dependent concentration.
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WB |
Use at an assay dependent concentration. Predicted molecular weight: 42 kDa.
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Flow Cyt (Intra) |
Use at an assay dependent concentration.
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mIHC |
Use at an assay dependent concentration.
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Notes |
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ICC/IF
Use at an assay dependent concentration. |
IHC-P
Use at an assay dependent concentration. |
WB
Use at an assay dependent concentration. Predicted molecular weight: 42 kDa. |
Flow Cyt (Intra)
Use at an assay dependent concentration. |
mIHC
Use at an assay dependent concentration. |
Target
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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
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Database links
- Entrez Gene: 423787 Chicken
- Entrez Gene: 515610 Cow
- Entrez Gene: 59 Human
- Entrez Gene: 11475 Mouse
- Entrez Gene: 733615 Pig
- Entrez Gene: 100009271 Rabbit
- Entrez Gene: 81633 Rat
- Omim: 102620 Human
see all -
Alternative names
- a actin antibody
- AAT6 antibody
- ACTA_HUMAN antibody
see all
Images
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Multiplex immunohistochemistry - Anti-alpha smooth muscle Actin antibody [SP171] - BSA and Azide free (ab242395)
Fluorescence multiplex immunohistochemical analysis of human liver tissue (formalin-fixed paraffin-embedded section). Panel A shows merged staining of ab195872 anti-Cytokeratin 19 stained on branch of bile ducts (magenta; Opal™690) at 1:8000 ( 0.127 μg/ml) [Panel B], ab275376 anti-Factor VIII stained on endothelial cells (red; Opal™570) at 1:1000 ( 0.457 μg/ml) [Panel C], and ab150301 anti-alpha smooth muscle Actin stained on smooth muscles (green; Opal™520) at 1:200 ( 0.14 μg/ml) [Panel C] on human liver. DAPI was used as a nuclear counter stain. Followed by Opal Polymer HRP Ms + Rb secondary. The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope. The section was incubated in three rounds of staining: in the order of ab195872 for 30 mins, ab275376 for 30 mins and ab150301 for 10 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins was used.
This data was developed using ab150301, the same antibody clone in a different buffer formulation.
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Immunocytochemistry/ Immunofluorescence - Anti-alpha smooth muscle Actin antibody [SP171] - BSA and Azide free (ab242395)This data was developed using the same antibody clone in a different buffer formulation (ab150301). ab150301 staining alpha smooth muscle Actin in SV40LT-SMC cells (positive control, top panel) and A431 cells (negative control, 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 ab150301 at 1μg/ml concentration and ab7291 (Mouse monoclonal to alpha Tubulin) at 1/1000 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor® 594) (ab150120) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems TCS SP8). -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody [SP171] - BSA and Azide free (ab242395)Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Rat colon tissue sections labeling alpha smooth muscle Actin with ab150301 at 1/200 dilution (0.26 µg/ml). Heat mediated antigen retrieval was performed Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 10 mins. Goat Anti-Rabbit & Mouse IgG (HRP) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab150301) -
Immunocytochemistry/ Immunofluorescence - Anti-alpha smooth muscle Actin antibody [SP171] - BSA and Azide free (ab242395)
Immunocytochemistry/ Immunofluorescence analysis of NIH/3T3 (mouse embryonic fibroblast) cells labeling alpha smooth muscle Actin with purified ab150301 at 1/100(1.65 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 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab150301). -
All lanes : Anti-alpha smooth muscle Actin antibody [SP171] (ab150301) at 1/130 dilution
Lane 1 : Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2 : ACTA2 knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 42 kDa
Observed band size: 42 kDaThis data was developed using the same antibody clone in a different buffer formulation (ab150301).
Lanes 1 - 2: Merged signal (red and green). Green - ab150301 observed at 42 kDa. Red - loading control, ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.
ab150301 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 ab150301 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4°C at a 1 in 130 Dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody [SP171] - BSA and Azide free (ab242395)Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Mouse colon tissue sections labeling alpha smooth muscle Actin with ab150301 at 1/200 dilution (0.26 µg/ml). Heat mediated antigen retrieval was performed Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 10 mins. Goat Anti-Rabbit & Mouse IgG (HRP) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab150301) -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody [SP171] - BSA and Azide free (ab242395)Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human colon tissue sections labeling alpha smooth muscle Actin with ab150301 at 1/200 dilution (0.26 µg/ml). Heat mediated antigen retrieval was performed Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 10 mins. Goat Anti-Rabbit & Mouse IgG (HRP) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab150301) -
Immunocytochemistry/ Immunofluorescence - Anti-alpha smooth muscle Actin antibody [SP171] - BSA and Azide free (ab242395)This data was developed using the same antibody clone in a different buffer formulation (ab150301). ab150301 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 ab150301 at 5μg/ml concentration and ab7291 (Mouse monoclonal to alpha Tubulin) at 1/1000 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor® 594) (ab150120) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems TCS SP8). -
Flow Cytometry (Intracellular) - Anti-alpha smooth muscle Actin antibody [SP171] - BSA and Azide free (ab242395)
Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma epithelial cell) cells labeling alpha smooth muscle Actin with purified ab150301 at 1/200 dilution (0.825µg/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (ab172730) / Black. Unlabeled control - Unlabelled cells / blue. This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab150301).
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Flow Cytometry (Intracellular) - Anti-alpha smooth muscle Actin antibody [SP171] - BSA and Azide free (ab242395)
Intracellular Flow Cytometry analysis of C6 (Rat glial tumor glial cell) cells labeling alpha smooth muscle Actin with purified ab150301 at 1/200 dilution (0.825µg/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (ab172730) / Black. Unlabeled control - Unlabelled cells / blue. This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab150301).
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Flow Cytometry (Intracellular) - Anti-alpha smooth muscle Actin antibody [SP171] - BSA and Azide free (ab242395)
Intracellular Flow Cytometry analysis of NIH/3T3 (Mouse embryonic fibroblast) cells labeling alpha smooth muscle Actin with purified ab150301 at 1/200 dilution (0.825µg/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (ab172730) / Black. Unlabeled control - Unlabelled cells / blue. This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab150301).
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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Datasheet download
References (0)
ab242395 has not yet been referenced specifically in any publications.