Overview

  • Product name
    Anti-beta Actin antibody [mAbcam 8226] - Loading Control (HRP)
    See all beta Actin primary antibodies
  • Description
    Mouse monoclonal [mAbcam 8226] to beta Actin - Loading Control (HRP)
  • Host species
    Mouse
  • Conjugation
    HRP
  • Tested applications
    Suitable for: WBmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Rabbit, Chicken, Cow, Dog, Human, Pig, Drosophila melanogaster, African green monkey, Chinese hamster
    Predicted to work with: Sheep, Horse, Guinea pig
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human beta Actin.

    Read Abcam's proprietary immunogen policy

  • Positive control
    • HeLa whole cell lysate, 3T3 whole cell lysate, Rat brain lysate

Properties

Applications

Our Abpromise guarantee covers the use of ab20272 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 1/5000. Detects a band of approximately 42 kDa (predicted molecular weight: 42 kDa).Can be blocked with Human beta Actin peptide (ab13772). Abcam strongly recommends blocking and incubation using 5% BSA as we have found that use of 5% milk seems to drastically reduce the intensity of the signal obtained. Please refer to WB image for further details.

Target

Images

  • All lanes : Anti-beta Actin antibody [mAbcam 8226] - Loading Control (HRP) (ab20272) at 1/5000 dilution

    Lane 1 : human testis lysate
    Lane 2 : human brain lysate
    Lane 3 : human tonsil lysate
    Lane 4 : mouse brain lysate
    Lane 5 : mouse spleen lysate

    Lysates/proteins at 20 µg per lane.

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 42 kDa


    Exposure time: 2 minutes
  • All lanes : Anti-beta Actin antibody [mAbcam 8226] - Loading Control (HRP) (ab20272) at 1/5000 dilution

    Lane 1 : HeLa whole cell lysate
    Lane 2 : 3T3 whole cell lysate
    Lane 3 : Rat brain lysate

    Lysates/proteins at 20 µg per lane.

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 42 kDa
    Observed band size: 42 kDa


    Exposure time: 30 seconds
  • Lanes 1-4 : Anti-beta Actin antibody [mAbcam 8226] - Loading Control (HRP) (ab20272) at 1/5000 dilution (Blocked in 5% BSA)
    Lanes 5-8 : Anti-beta Actin antibody [mAbcam 8226] - Loading Control (HRP) (ab20272) at 1/5000 dilution (Blocked in 5% MILK)

    Lanes 1 & 5 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lanes 2 & 6 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
    Lanes 3 & 7 : NIH 3T3 whole cell lysate (ab7179)
    Lanes 4 & 8 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 42 kDa
    Observed band size: 42 kDa


    Exposure time: 1 minute


    The membrane 1-4 was blocked in 5% BSA (1 hour). The membrane 5-8 was blocked in 5% MILK (1 hour). Abcam routinely uses 5% BSA to block, however following recent customer feedback our labs investigated the effect of 5% milk blocking. We can now confirm that milk is not a suitable blocking agent for this antibody and significantly decreases the signal on the membrane.

References

This product has been referenced in:
  • Khan NA  et al. Waterpipe smoke and e-cigarette vapor differentially affect circadian molecular clock gene expression in mouse lungs. PLoS One 14:e0211645 (2019). Read more (PubMed: 30811401) »
  • Kosgodage US  et al. Cannabidiol Affects Extracellular Vesicle Release, miR21 and miR126, and Reduces Prohibitin Protein in Glioblastoma Multiforme Cells. Transl Oncol 12:513-522 (2019). Read more (PubMed: 30597288) »
See all 133 Publications for this product

Customer reviews and Q&As

11-20 of 20 Abreviews or Q&A

Application
Western blot
Sample
Mouse Tissue lysate - whole (liver)
Loading amount
40 µg
Specification
liver
Gel Running Conditions
Reduced Denaturing
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 Sep 21 2012

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

Abcam user community

Verified customer

Submitted Nov 10 2009

Application
Western blot
Sample
Human Cell lysate - whole cell (HEK-293)
Loading amount
50000 cells
Specification
HEK-293
Blocking step
Western Breeze Kit as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: Kit

Dr. Alan Hendrick

Verified customer

Submitted Apr 17 2007

Application
Western blot
Sample
Mouse Tissue lysate - whole (Brain)
Loading amount
50 µg
Specification
Brain
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%

Abcam user community

Verified customer

Submitted Dec 01 2006

Application
Western blot
Sample
African Green Monkey Cell lysate - whole cell (Cos7 transfected with pCMV-Nle-Myc)
Loading amount
1e+006 cells
Specification
Cos7 transfected with pCMV-Nle-Myc
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%

Abcam user community

Verified customer

Submitted May 05 2006

Application
Western blot
Sample
Cow Cell lysate - whole cell (retinal endothelial cell)
Specification
retinal endothelial cell
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%

Abcam user community

Verified customer

Submitted Feb 08 2006

Application
Western blot
Sample
Mouse Cell lysate - whole cell (retinal pigment epithelium)
Specification
retinal pigment epithelium
Blocking step
Milk as blocking agent for 45 minute(s) · Concentration: 5%

Abcam user community

Verified customer

Submitted Jan 30 2006

Application
Western blot
Sample
Human Cell lysate - whole cell (retinal pigment epithelium)
Specification
retinal pigment epithelium
Blocking step
Milk as blocking agent for 45 minute(s) · Concentration: 5%

Abcam user community

Verified customer

Submitted Jan 30 2006

Application
Western blot
Sample
Dog Cell lysate - whole cell (Kidney epithelial cell line)
Specification
Kidney epithelial cell line
Blocking step
Milk as blocking agent for 45 minute(s) · Concentration: 5%

Abcam user community

Verified customer

Submitted Jan 30 2006

Application
Western blot
Sample
Rat Cell lysate - whole cell (rat dermal cell)
Specification
rat dermal cell
Blocking step
Milk as blocking agent for 45 minute(s) · Concentration: 5%

Abcam user community

Verified customer

Submitted Jan 30 2006

11-20 of 20 Abreviews or Q&A

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