Anti-beta Actin antibody [mAbcam 8226] - Loading Control (HRP) (ab20272)

Overview

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

Anti-beta Actin antibody [mAbcam 8226] - Loading Control (HRP) 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)

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
    Lane 3 : NIH 3T3 whole cell lysate (ab7179)
    Lane 4 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate
    Lane 5 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 6 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
    Lane 7 : NIH 3T3 whole cell lysate (ab7179)
    Lane 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 for Anti-beta Actin antibody [mAbcam 8226] - Loading Control (HRP) (ab20272)

This product has been referenced in:
  • Baran-Gale J  et al. An integrative transcriptomics approach identifies miR-503 as a candidate master regulator of the estrogen response in MCF-7 breast cancer cells. RNA 22:1592-603 (2016). WB ; Human . Read more (PubMed: 27539783) »
  • Guzyk MM  et al. Poly(ADP-Ribose) Polymerase-1 (PARP-1) Inhibitors Reduce Reactive Gliosis and Improve Angiostatin Levels in Retina of Diabetic Rats. Neurochem Res 41:2526-2537 (2016). Read more (PubMed: 27255598) »

See all 50 Publications for this product

Product Wall

Application Western blot
Sample Mouse Tissue lysate - whole (Spleen)
Gel Running Conditions Reduced Denaturing (12%)
Loading amount 20 µg
Specification Spleen
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
Username

Dr. Sergi Bayod

Verified customer

Submitted Mar 24 2017

Application Western blot
Loading amount 30 µg
Gel Running Conditions Reduced Denaturing
Sample Rat Cell lysate - whole cell (h9c2)
Specification h9c2
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: rt°C
Username

Abcam user community

Verified customer

Submitted Jan 02 2014

Application Western blot
Loading amount 30 µg
Gel Running Conditions Reduced Denaturing
Sample Mouse Cell lysate - whole cell (MEF)
Specification MEF
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: rt°C
Username

Abcam user community

Verified customer

Submitted Jan 02 2014

Application Western blot
Loading amount 30 µg
Gel Running Conditions Reduced Denaturing
Sample Human Cell lysate - whole cell (hek293)
Specification hek293
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: rt°C
Username

Abcam user community

Verified customer

Submitted Jan 02 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
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
Username

Abcam user community

Verified customer

Submitted Sep 21 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
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
Username

Abcam user community

Verified customer

Submitted Nov 10 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
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
Username

Dr. Alan Hendrick

Verified customer

Submitted Apr 17 2007

Abcam guarantees this product to work in the species/application used in this Abreview.
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%
Username

Abcam user community

Verified customer

Submitted Dec 01 2006

Abcam guarantees this product to work in the species/application used in this Abreview.
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%
Username

Abcam user community

Verified customer

Submitted May 05 2006

Abcam guarantees this product to work in the species/application used in this Abreview.
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%
Username

Abcam user community

Verified customer

Submitted Feb 08 2006

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