Overview

  • Product nameAnti-beta Actin antibody [BA3R]
    See all beta Actin primary antibodies
  • Description
    Mouse monoclonal [BA3R] to beta Actin
  • SpecificityRecognizes native and denatured forms of ß-Actin (~42kDa)
  • Tested applicationsELISA, Dot Blot, WB, Flow Cyt, ICC/IF, IPmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Rabbit, Chicken, Human
    Predicted to work with: Sheep, Horse, Guinea pig, Hamster, Cow, Pig, Chimpanzee, Cynomolgus Monkey, Japanese ricefish, Orangutan
  • Immunogen

    Synthetic peptide corresponding to amino acids 2 - 15 (DDDIAALVVDNGSG) of Human beta Actin cytoplasmic domain (slightly modified)

  • Positive control
    • 293 cell lysate. This antibody gave a positive result when used in the following methanol fixed cell line: HeLa.

Properties

Applications

Our Abpromise guarantee covers the use of ab92436 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
ELISA Use at an assay dependent concentration.
Dot Blot Use at an assay dependent concentration.
WB 1/1000 - 1/3000. Predicted molecular weight: 42 kDa.
Flow Cyt Use 0.1-1µg for 106 cells.
ICC/IF Use a concentration of 10 µg/ml.
IP Use a concentration of 5 µg/ml.

Target

Anti-beta Actin antibody [BA3R] images

  • beta Actin was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Mouse monoclonal to beta Actin and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).

    The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.

    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab92436.

    Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/20,000 dilution.

    Band: 42kDa; beta Actin

  • ICC/IF image of ab92436 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab92436 at 10 µg/mL overnight at +4°C. The secondary antibody (green) was a goat anti-mouse DyLight® 488 (IgG; H+L) (ab96879) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • Overlay histogram showing HeLa cells stained with ab92436 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab92436, 0.1μg/1x106 cells) for 30 min at 22°C. The secondary antibody used was a goat anti-mouse Alexa Fluor® 488 (IgG; H+L) (ab150113) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in HeLa cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

  • All lanes : Anti-beta Actin antibody [BA3R] (ab92436) at 1/1000 dilution

    Lane 1 : Human at 20 µg/ml
    Lane 2 : Mouse at 20 µg/ml
    Lane 3 : Rat at 20 µg/ml
    Lane 4 : Rabbit at 20 µg/ml


    Predicted band size : 42 kDa
  • All lanes : Anti-beta Actin antibody [BA3R] (ab92436) at 1/1000 dilution

    Lane 1 : 293 cell lysate at 3 µg
    Lane 2 : 293 cell lysate at 10 µg
    Lane 3 : 293 cell lysate at 30 µg


    Predicted band size : 42 kDa

References for Anti-beta Actin antibody [BA3R] (ab92436)

ab92436 has not yet been referenced specifically in any publications.

Product Wall

There are currently no Abreviews or Questions for ab92436.
Please use the links above to contact us or submit feedback about this product.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"