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beta Actin Antibody
beta Actin Monoclonal Antibody for FC, IF, IHC, IP, WB, ELISA
Host / Isotype
Mouse / IgG2b
Reactivity
human, mouse, rat, hamster, zebrafish, monkey and More (10)
Applications
FC, IF, IHC, IP, WB, ELISA
Conjugate
Unconjugated
1027
CloneNo.
2D4H5
Cat no : 66009-1-Ig
Synonyms
ACTB, Actin, actin, beta, Actin, cytoplasmic 1, B actin, Beta actin, F actin, PS1TP5BP1, β actin
Validation Data Gallery
Tested Applications
| Positive WB detected in | multi-cells, Jurkat cells, A549 cells, CHO cells, HeLa cells |
| Positive IP detected in | HeLa cells |
| Positive IHC detected in | human 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 in | MDCK cells, HepG2 cells |
| Positive FC detected in | HeLa cells |
Recommended dilution
| Application | Dilution |
|---|---|
| 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 : 1:200-1:1500 |
| Sample-dependent, check data in validation data gallery | |
Published Applications
| WB | See 979 publications below |
| IHC | See 1 publications below |
| IF | See 5 publications below |
Product Information
The 66009-1-Ig immunogen is Fusion Protein beta Actin protein expressed in E. coli.
| Reactivity | human, mouse, rat, hamster, zebrafish, monkey |
| Cited Species | broiler chick, chick, chicken, chicks, common carp, common carps, cow, dog, Drosophila, Duck |
| 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 number | NM_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 Conditions | Store at -20oC. 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-Ig | Download protocol |
| IHC protocol for beta Actin antibody 66009-1-Ig | Download protocol |
| IF protocol for beta Actin antibody 66009-1-Ig | Download protocol |
| IP protocol for beta Actin antibody 66009-1-Ig | Download protocol |
| FC protocol for beta Actin antibody 66009-1-Ig | Download protocol |
| Standard Protocols | |
|---|---|
| Click here to view our Standard Protocols |
Publications
| Species | Application | Title |
|---|---|---|
Proc Natl Acad Sci U S A Cell-autonomous adiposity through increased cell surface GLUT4 due to ankyrin-B deficiency. | ||
Evid Based Complement Alternat Med Tetrandrine-Induced Autophagy in MDA-MB-231 Triple-Negative Breast Cancer Cell through the Inhibition of PI3K/AKT/mTOR Signaling. | ||
Cell Cycle CDK11p110 plays a critical role in the tumorigenicity of esophageal squamous cell carcinoma cells and is a potential drug target. | ||
Nat Commun Intestinal non-canonical NFκB signaling shapes the local and systemic immune response. | ||
Transl Neurosci Protective Effect of Extract of Bletilla Striata on Isoflurane Induced Neuronal Injury By Altering PI3K/Akt Pathway. | ||
Biotechnol Prog Apoptosis and cell cycle arrest of hepatocellular carcinoma spheroids treated by an alternating electric field. |
Reviews
The reviews below have been submitted by verified Proteintech customers who received an incentive forproviding their feedback.
FH Magda Czepnik (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.
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FH Elisa Villalobos (10-21-2019) | The antibody works very nicely, very clean and I used only 10 uL of protein.
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FH Tracy Chih-Ting Koubkova Yu (09-24-2019) | I used this antibody for Hydra actin immunostaining. The outcome is quite good.
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FH Rui Yan (09-02-2019) | Very specific and strong labeling in whole cell lysates.
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FH Huai-Chin Chiang (06-19-2019) | Very good anitbody. Works well in Western for various type of cells.
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FH Francesco Gubieli (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
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FH Kyosuke Kazama (06-05-2019) | This antibody works very well for Western blot using vascular tissue or cell lysates.
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