Recombinant

Recombinant Anti-beta Actin antibody [EPR21241] (ab213262)

Overview

  • 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.

  • Tested applications

    Suitable for: WB, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Human
    Predicted to work with: Cow, Dog, Pig
  • 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
  • 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

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

  • 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
    modifications

    ISGylated.
  • Cellular localization

    Cytoplasm > cytoskeleton. Localized in cytoplasmic mRNP granules containing untranslated mRNAs.
  • Information by UniProt
  • Database links

  • Alternative names

    • A26C1A antibody
    • A26C1B antibody
    • ACTB antibody
    • ACTB_HUMAN antibody
    • Actin beta antibody
    • Actin cytoplasmic 1 antibody
    • Actin, cytoplasmic 1, N-terminally processed antibody
    • Actx antibody
    • b actin antibody
    • b-actin antibody
    • Beta cytoskeletal actin antibody
    • Beta-actin antibody
    • BRWS1 antibody
    • E430023M04Rik antibody
    • Melanoma X actin antibody
    • MGC128179 antibody
    • PS1TP5 binding protein 1 antibody
    • PS1TP5BP1 antibody
    see all

Images

  • 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).

  • 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 kDa



    ab213262 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.

  • 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).

References

ab213262 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

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1-2 of 2 Abreviews

Application
Western blot
Sample
Mouse Cell lysate - whole cell (Macropahges)
Gel Running Conditions
Reduced Denaturing
Loading amount
10 µg
Specification
Macropahges
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Jun 07 2019

Application
Western blot
Sample
Human Cell lysate - whole cell (Endothelial cell)
Gel Running Conditions
Reduced Denaturing
Loading amount
10 µg
Specification
Endothelial cell
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Apr 05 2019

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