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) »
  • Soriano-Úbeda C  et al. Manipulation of bicarbonate concentration in sperm capacitation media improvesin vitro fertilisation output in porcine species. J Anim Sci Biotechnol 10:19 (2019). Read more (PubMed: 30899459) »
See all 149 Publications for this product

Customer reviews and Q&As

1-10 of 20 Abreviews or Q&A

Application
Western blot
Sample
Rat Tissue lysate - whole (liver)
Gel Running Conditions
Reduced Denaturing (12%)
Loading amount
30 µg
Specification
liver
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Abcam user community

Verified customer

Submitted Mar 13 2019

Application
Western blot
Sample
Rat Cell lysate - whole cell (Neruo cell)
Gel Running Conditions
Reduced Denaturing (12 %)
Loading amount
20 µg
Specification
Neruo cell
Blocking step
BSA as blocking agent for 20 minute(s) · Concentration: 5% · Temperature: 27°C

Miss. Ji Yeon Lee

Verified customer

Submitted Jan 28 2019

Application
Western blot
Sample
Rat Cell lysate - whole cell (Cardiomyocytes)
Gel Running Conditions
Reduced Denaturing
Loading amount
25 µg
Specification
Cardiomyocytes
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 Dec 29 2017

Application
Western blot
Sample
Human Cell lysate - whole cell (HEK293)
Gel Running Conditions
Reduced Denaturing (15% PAGE)
Loading amount
50 µg
Specification
HEK293
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 Dec 29 2017

Application
Western blot
Sample
Monkey Cell lysate - whole cell (Heart)
Gel Running Conditions
Reduced Denaturing
Loading amount
50 µg
Specification
Heart
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 Dec 29 2017

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

Abcam user community

Verified customer

Submitted Dec 20 2017

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

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

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

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

Abcam user community

Verified customer

Submitted Jan 02 2014

1-10 of 20 Abreviews or Q&A

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