Validation Data Gallery
Tested Applications
| Positive WB detected in | HeLa cells, A549 cells, Jurkat cells, mouse pancreas, rat brain, rat pancreas, HEK-293 cells, HepG2 cells, K-562 cells, HSC-T6 cells, NIH/3T3 cells, 4T1 cells, CHO cells, HSC-T6 cells. NIH/3T3 cells, Pig brain, Rabbit brain, Mouse brain, Chicken brain |
| 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-P detected in | mouse brain tissue |
| Positive IF/ICC detected in | MDCK cells, HeLa cells |
| Positive FC (Intra) detected in | HeLa cells |
Recommended dilution
| Application | Dilution |
|---|---|
| Western Blot (WB) | WB : 1:20000-1:100000 |
| Immunoprecipitation (IP) | IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate |
| Immunohistochemistry (IHC) | IHC : 1:20-1:2000 |
| Immunofluorescence (IF)-P | IF-P : 1:500-1:2000 |
| Immunofluorescence (IF)/ICC | IF/ICC : 1:500-1:2000 |
| Flow Cytometry (FC) (INTRA) | FC (INTRA) : 0.40 ug per 10^6 cells in a 100 µl suspension |
| It is recommended that this reagent should be titrated in each testing system to obtain optimal results. | |
| Sample-dependent, Check data in validation data gallery. | |
Product Information
66009-1-Ig targets Beta Actin in WB, IHC, IF/ICC, IF-P, FC (Intra), IP, CoIP, ChIP, ELISA applications and shows reactivity with human, mouse, rat, pig, rabbit, canine, monkey, chicken, zebrafish, hamster samples.
| Tested Reactivity | human, mouse, rat, pig, rabbit, canine, monkey, chicken, zebrafish, hamster |
| Cited Reactivity | human, mouse, rat, monkey, chicken, sheep, goat, arabidopsis, golden hamsters, aedes albopictus |
| Host / Isotype | Mouse / IgG2b |
| Class | Monoclonal |
| Type | Antibody |
| Immunogen | Recombinant protein 相同性解析による交差性が予測される生物種 |
| Full Name | actin, beta |
| Calculated molecular weight | 42 kDa |
| Observed molecular weight | 42 kDa |
| GenBank accession number | NM_001101 |
| Gene Symbol | Beta Actin |
| Gene ID (NCBI) | 60 |
| RRID | AB_2687938 |
| Conjugate | Unconjugated |
| Form | Liquid |
| Purification Method | Protein A purification |
| UNIPROT ID | P60709 |
| Storage Buffer | PBS with 0.02% sodium azide and 50% glycerol pH 7.3. |
| Storage Conditions | Store 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-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 |
|---|---|---|
Cell Res Mannose antagonizes GSDME-mediated pyroptosis through AMPK activated by metabolite GlcNAc-6P | ||