Recombinant Anti-beta Actin antibody [EPR21241] (ab213262)
- Datasheet
- References
- Protocols
Overview
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Product name
Anti-beta Actin antibody [EPR21241]
See all beta Actin primary antibodies -
Description
Rabbit monoclonal [EPR21241] to beta Actin -
Host species
Rabbit -
Specificity
In-house testing showed ab213262 to react with gamma actin in western blot.
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Tested applications
Suitable for: WB, ICC/IFmore details -
Species reactivity
Reacts with: Mouse, Human
Predicted to work with: Cow, Dog, Pig
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Immunogen
Synthetic peptide aa 1-100.
Database link: P60709 -
Positive control
- ICC/IF: Methanol-fixed HeLa and NIH3T3 cells. WB: HAP1, MEF1 and NIH3T3 cells
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General notes
This product was made using synthetic libraries and phage display technology.
This antibody is a recombinant chimeric antibody. Rabbit chimeric monoclonal antibody (Human Fab/ Rabbit Fc).
If this antibody is used as a loading control in WB and if the signal of the gamma actin is close to the protein of interest to be detected, we recommend cutting the membrane between the expected signals of beta actin and gamma actin and to stain the two parts containing the loading control and the protein of interest separately.
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 long term. Avoid freeze / thaw cycle. -
Concentration information loading... -
Purity
Immunogen affinity purified -
Clonality
Monoclonal -
Clone number
EPR21241 -
Isotype
IgG1 -
Research areas
Associated products
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Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 555) (ab150078)
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) (ab150079)
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) (ab150080)
- Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
- Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96899)
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Recombinant Protein
Applications
Our Abpromise guarantee covers the use of ab213262 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 |
|---|---|---|
| WB | Use a concentration of 1 µg/ml. Predicted molecular weight: 42 kDa. | |
| ICC/IF | Use a concentration of 0.05 - 0.5 µg/ml. |
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 ACTB are a cause of dystonia juvenile-onset (DYTJ) [MIM:607371]. DYTJ is a form of dystonia with juvenile onset. Dystonia is defined by the presence of sustained involuntary muscle contraction, often leading to abnormal postures. DYTJ patients manifest progressive, generalized, dopa-unresponsive dystonia, developmental malformations and sensory hearing loss. -
Sequence similarities
Belongs to the actin family. -
Post-translational
modificationsISGylated. -
Cellular localization
Cytoplasm > cytoskeleton. Localized in cytoplasmic mRNP granules containing untranslated mRNAs. - Information by UniProt
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Database links
- Entrez Gene: 280979 Cow
- Entrez Gene: 403580 Dog
- Entrez Gene: 60 Human
- Entrez Gene: 11461 Mouse
- Entrez Gene: 414396 Pig
- Omim: 102630 Human
- SwissProt: P60712 Cow
- SwissProt: O18840 Dog
see all -
Alternative names
- A26C1A antibody
- A26C1B antibody
- ACTB antibody
see all
Images
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![Immunocytochemistry/ Immunofluorescence - Anti-beta Actin antibody [EPR21241] (ab213262)](https://www.abcam.com/ps/products/213/ab213262/Images/ab213262-320188-anti-beta-actin-antibody-epr21241-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-beta Actin antibody [EPR21241] (ab213262)ab213243 staining beta-Actin in NIH3T3 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 ab213262 at 0.5ugml then detected with an Alexa Fluor® 488 goat anti-rabbit secondary antibody (ab150081) at a 1/1000 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue), and ab195889, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 594), at a 1/250 dilution (shown in red).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
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![Western blot - Anti-beta Actin antibody [EPR21241] (ab213262)](https://www.abcam.com/ps/products/213/ab213262/Images/ab213262-337175-anti-beta-actin-antibody-epr21241-western-blot.jpg)
Western blot - Anti-beta Actin antibody [EPR21241] (ab213262)All lanes :
Lane 1 : HAP1 whole cell lysate at 20 µg
Lane 2 : HAP1 ACTNB knockout whole cell lysate at 20 µg
Lane 3 : MEF1 whole cell lysate at 20 µg
Lane 4 : NIH3T3 whole cell lysate at 20 µg
Lane 5 :Recombinant Human gamma Actin protein (ab157841)
Performed under reducing conditions.
Predicted band size: 42 kDaab213262 was shown to react with ACTNB (beta actin) when tested against HAP1 wild-type and HAP1 ACTNB (beta actin) knockout cells. ACTNB wild-type and knockout samples were subjected to SDS-PAGE and incubated with ab213262. Incomplete reduction of signal was seen in the ACTNB (beta actin) knockout samples at beta actin's expected molecular weight. SDS-PAGE of gamma actin recombinant protein and incubation with ab213262 showed cross-reactivity to gamma actin.
ab213262 and ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at a 1ug/ml and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
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![Immunocytochemistry/ Immunofluorescence - Anti-beta Actin antibody [EPR21241] (ab213262)](https://www.abcam.com/ps/products/213/ab213262/Images/ab213262-320187-anti-beta-actin-antibody-epr21241-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-beta Actin antibody [EPR21241] (ab213262)ab213243 staining beta-Actin in HeLa 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 ab213262 at 0.01ugml then detected with an Alexa Fluor® 488 goat anti-rabbit secondary antibody (ab150081) at a 1/1000 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue), and ab195889, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 594), at a 1/250 dilution (shown in red).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Datasheets and documents
References
ab213262 has not yet been referenced specifically in any publications.
