• Featured Product
  • KD/KO Validated

Beta Actin Monoclonal antibody

Beta Actin Monoclonal Antibody for FC, IF, IHC, IP, WB,ELISA

Host / Isotype

Mouse / IgG2b

Reactivity

human, mouse, rat, hamster, zebrafish, monkey, dog and More (9)

Applications

WB, IP, IHC, IF, FC, CoIP,ELISA

Conjugate

Unconjugated

CloneNo.

2D4H5

Cat no : 66009-1-Ig

Synonyms

ACTB, Actin, actin, beta, Actin, cytoplasmic 1, B actin, Beta actin, F actin, PS1TP5BP1, β actin


"Beta Actin Antibodies" Comparison

View side-by-side comparison of Beta Actin antibodies from other vendors to find the one that best suits your research needs.
  • WB analysis of multi-cells using 66009-1-Ig

    WB analysis of multi-cells using 66009-1-Ig

    Western blot analysis of beta-actin in various tissues and cell lines using Proteintech antibody 66009-1-Ig at a dilution of 1:20000. (Exposure time: 10 seconds).

  • IF Staining of MDCK using 66009-1-Ig

    IF Staining of MDCK using 66009-1-Ig

    Immunofluorescent analysis of (-20°C Methanol) fixed MDCK cells using Beta Actin antibody (66009-1-Ig, Clone: 2D4H5 ) at dilution of 1:1000 and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L).

  • WB analysis using 66009-1-Ig

    WB analysis using 66009-1-Ig

    Various lysates were subjected to SDS PAGE followed by western blot with 66009-1-Ig (beta Actin antibody) at dilution of 1:8000 incubated at room temperature for 1.5 hours.

  • WB analysis of A549 cells using 66009-1-Ig

    WB analysis of A549 cells using 66009-1-Ig

    A549 cells (shcontrol and shRNA of Beta Actin) were subjected to SDS PAGE followed by western blot with 66009-1-Ig (Mouse anti Pan-actin antibody) at dilution of 1:10000.

  • IP experiment of HeLa using 66009-1-Ig

    IP experiment of HeLa using 66009-1-Ig

    IP Result of anti-beta actin (IP:66009-1-Ig, 3ug; Detection:66009-1-Ig 1:10000) with HeLa cells lysate 3100ug.

  • FC experiment of HeLa using 66009-1-Ig

    FC experiment of HeLa using 66009-1-Ig

    1X10^6 HeLa cells were stained with 0.05 ug Anti-Human Beta Actin (66009-1-Ig, Clone:2D4H5) and FITC-Goat anti-mouse IgG at dilution 1:100 (red), or stained with 0.05 ug isotype control and FITC-Goat anti-mouse IgG at dilution 1:100 (blue). Cells were fixed with 4% PFA and permeabilized with 0.1% TritonX-100.

  • IHC staining of human kidney using 66009-1-Ig

    IHC staining of human kidney using 66009-1-Ig

    Immunohistochemical analysis of paraffin-embedded human kidney using 66009-1-Ig at dilution of 1:500 (under 40x lens).


Tested Applications

Positive WB detected inmulti-cells, Jurkat cells, A549 cells, CHO cells, HeLa cells
Positive IP detected inHeLa cells
Positive IHC detected inhuman kidney tissue, human brain tissue, human colon cancer tissue, human heart tissue
Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
Positive IF detected inMDCK cells, HepG2 cells
Positive FC detected inHeLa cells
Planning an IF experiment? We recommend our CoraLite®488, CoraLite®555 and CoraLite®594 conjugated versions of this antibody.

Recommended dilution

ApplicationDilution
Western Blot (WB)WB : 1:5000-1:50000
Immunoprecipitation (IP)IP : 0.5-4.0 ug for IP and 1:5000-1:50000 for WB
Immunohistochemistry (IHC)IHC : 1:20-1:2000
Immunofluorescence (IF)IF : 1:500-1:2000
Sample-dependent, check data in validation data gallery

Product Information

The immunogen of 66009-1-Ig is Beta Actin Fusion Protein expressed in E. coli.

Tested Reactivity human, mouse, rat, hamster, zebrafish, monkey, dog
Cited Reactivity Aedes albopictus, Arabidopsis, bovine, canine, chicken, Cyprinus carpio, dog, Drosophila, duck, fish
Host / Isotype Mouse / IgG2b
Class Monoclonal
Type Antibody
Immunogen Fusion Protein
Full Name actin, beta
Calculated molecular weight 42 kDa
Observed molecular weight 42 kDa
GenBank accession numberNM_001101
Gene symbol ACTB
Gene ID (NCBI) 60
Conjugate Unconjugated
Form Liquid
Purification Method Protein A purification
Storage Buffer PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
Storage ConditionsStore at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20oC storage.

Background Information

Actins are highly conserved globular proteins that are involved in various types of cell motility and are ubiquitously expressed in all eukaryotic cells. At least six isoforms of actins are known in mammals and other vertebrates. Beta-actin (or β-actin) is a cytoskeletal actin isoform, highly conserved and ubiquitously expressed in eukaryotic organisms, making it a commonly used "housekeeping gene" in many laboratory techniques. It has been widely used as the internal control in RT-PCR and Western Blotting. The isotype of this antibody is IgG2b.

 

What is the molecular weight of beta-actin?

42 kDa

 

What is the function of beta-actin?

Actins are a key part of the cytoskeleton and are involved in cell motility, structure, and signaling, with beta-actin as part of the contractile apparatus. Beta-actin is located in the cytoplasm of non-muscle cells, particularly where the plasma membrane meets the cytoskeleton (PMID: 1993736) and is found co-distributed with alpha-actin (PMID: 9490838).

 

What are the applications for beta-actin?

As it is ubiquitously and constitutively expressed, beta-actin is a useful loading control in techniques such as RT-PCR and Western Blots to measure baseline expression and degradation. Beta-actin can be used as a reference gene to normalize mRNA or protein levels in quantitative studies of expression. Although some evidence suggests variation in tissue types and developing embryos (PMID: 23099774) if expression is stable across conditions, beta-actin is still considered by many to be a suitable gene for normalization.

Beyond its role as a housekeeping gene, it has been shown that beta-actin associates with endothelial nitric-oxide synthase (eNOS) and can therefore regulate nitric oxide activity in endothelial cells (PMID: 12734108).

 

What are the post-transcriptional modifications (PTMs) to beta actin?

Alterations to actin proteins can modulate their properties and function. While small molecules are known to alter actin, post-translational modifications such as phosphorylation and acetylation can also affect actin (PMID: 23195437). Modifications to actin have been associated with diseases including but not limited to age-related macular degeneration (PMID: 17652714), cardiovascular disease (PMID: 21232667), and Alzheimer's disease (PMID: 11246152). 


For murine tissue sample, conjugated antibody (HRP-66009) or rabbit antibody (20536-1-AP) is preferable.



Protocols

Product Specific Protocols
WB protocol for Beta Actin antibody 66009-1-IgDownload protocol
IHC protocol for Beta Actin antibody 66009-1-IgDownload protocol
IF protocol for Beta Actin antibody 66009-1-IgDownload protocol
IP protocol for Beta Actin antibody 66009-1-IgDownload protocol
FC protocol for Beta Actin antibody 66009-1-IgDownload protocol
Standard Protocols
Click here to view our Standard Protocols

Publications

SpeciesApplicationTitle
mouseWB

Proc Natl Acad Sci U S A

Cell-autonomous adiposity through increased cell surface GLUT4 due to ankyrin-B deficiency.

Authors - Damaris N Lorenzo
ratWB

Mol Med Rep

Function of BRD4 in the pathogenesis of high glucose‑induced cardiac hypertrophy.

Authors - Qian Wang
ratWB

J Cell Mol Med

Nucleoporin 107 facilitates the nuclear export of Scn5a mRNA to regulate cardiac bioelectricity.

Authors - Yi Guan
mouseWB

J Diabetes

Inhibition of HMGB1 attenuates podocyte apoptosis and epithelial-to-mesenchymal transition (EMT) by regulating autophagy flux.

Authors - Juan Jin
humanWB

Theranostics

Oncogenic MSH6-CXCR4-TGFB1 Feedback Loop: A Novel Therapeutic Target of Photothermal Therapy in Glioblastoma Multiforme.

Authors - Yaodong Chen
pigWB

Arch Virol

Porcine circovirus type 2 ORF5 protein induces endoplasmic reticulum stress and unfolded protein response in porcine alveolar macrophages.

Authors - Yueling Ouyang

Reviews

The reviews below have been submitted by verified Proteintech customers who received an incentive forproviding their feedback.


FH

Eric (Verified Customer) (06-25-2021)

We ordered this actin antibody to detect actin in the cell lysates during the western blot, and it works very well, and the antibody is very sensitive. We highly recommend it.

  • Applications: Western Blot
  • Cell Tissue Type: human macrophages
Beta Actin Antibody Western Blot validation ( dilution) in human macrophages (Cat no:66009-1-Ig)
FH

Elisa (Verified Customer) (06-14-2021)

This antibody work really well in cells and tissues (mouse). I have also used in human skeletal muscle with good results as well.

  • Applications: Western Blot
  • Primary Antibody Dilution: 1:5000
  • Cell Tissue Type: C2C12, Heart, skeletal muscle, adipose, kidney, liver
Beta Actin Antibody Western Blot validation (1:5000 dilution) in C2C12, Heart, skeletal muscle, adipose, kidney, liver (Cat no:66009-1-Ig)
FH

Ambra (Verified Customer) (01-20-2021)

Works nicely.Used on Human brain lysates (temporal cortex).

  • Applications: Western Blot
  • Primary Antibody Dilution: 1:2000
  • Cell Tissue Type: Human brain lysate
Beta Actin Antibody Western Blot validation (1:2000 dilution) in Human brain lysate (Cat no:66009-1-Ig)
FH

V (Verified Customer) (01-12-2021)

Very good product; signal is good

  • Applications: Western Blot
FH

Yu Siong (Verified Customer) (07-30-2020)

Generate very consistent result for my porcine brain endothelial cells. The antibody at dilution of 1:5000 can be reused for multiple times.

  • Applications: Western Blot
  • Primary Antibody Dilution: 1:5000
  • Cell Tissue Type: porcine brain endothelial cells
Beta Actin Antibody Western Blot validation (1:5000 dilution) in porcine brain endothelial cells (Cat no:66009-1-Ig)
FH

Magda (Verified Customer) (12-31-2019)

I needed to run a housekeeping gene control for my protein expression test. I was testing HEK293T cell lysates. My Lab did not have a proper AB, but a colleague from a lab next door borrowed me a sample from Proteintech. I was very happy with the results, they were consistent among my triplicate. I would purchase this AB but as a HRP conjugate, as it saves more time and provides cleaner results.

  • Applications: Western Blot,
  • Cell Tissue Type: HEK293T
FH

Elisa (Verified Customer) (10-21-2019)

The antibody works very nicely, very clean and I used only 10 uL of protein.

  • Applications: Western Blot,
  • Primary Antibody Dilution: 1:5000
  • Cell Tissue Type: Cell line C2C12
FH

Tracy Chih-Ting (Verified Customer) (09-24-2019)

I used this antibody for Hydra actin immunostaining. The outcome is quite good.

  • Applications: Immunofluorescence,
  • Primary Antibody Dilution: 1:500
FH

Rui (Verified Customer) (09-02-2019)

Very specific and strong labeling in whole cell lysates.

  • Applications: Western Blot,
  • Primary Antibody Dilution: 1:5000
  • Cell Tissue Type: Cos-7
FH

Huai-Chin (Verified Customer) (06-19-2019)

Very good anitbody. Works well in Western for various type of cells.

  • Applications: Western Blot,
  • Primary Antibody Dilution: 1:3000
  • Cell Tissue Type: Primary cells, various cell lines
FH

Francesco (Verified Customer) (06-12-2019)

in the testing staining it kind of worked, even if with higher dilution it could work better.anyway further test should be done to confirm it.if results are reproducible it might be a valid alternative to more expensive antibody

  • Applications: Immunohistochemistry,
  • Primary Antibody Dilution: 1/1500
  • Cell Tissue Type: Brain cells
FH

Kyosuke (Verified Customer) (06-05-2019)

This antibody works very well for Western blot using vascular tissue or cell lysates.

  • Applications: Western Blot,
  • Primary Antibody Dilution: 1/1000
  • Cell Tissue Type: aorta