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beta-Actin Recombinant Protein Antigen

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Product Details

Summary
Reactivity HuSpecies Glossary
Applications AC

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beta-Actin Recombinant Protein Antigen Summary

Description
A recombinant protein antigen with a N-terminal His6-ABP tag corresponding to human beta-Actin

Source: E. coli

Amino Acid Sequence: PKANREKMTQIMFETFNTPAMYVAIQ

Fusion Tag: N-terminal His6ABP (ABP = Albumin Binding Protein derived from Streptococcal Protein G)

This product is intended to be used as a blocking antigen for antibody competition assays. Any other use of this antigen is done at the risk of the user. The use of this product for commercial production is strictly prohibited. Please contact technical support if you have any questions.

Source
E. coli
Protein/Peptide Type
Recombinant Protein Antigen
Gene
ACTB
Purity
>80% by SDS-PAGE and Coomassie blue staining

Applications/Dilutions

Dilutions
  • Antibody Competition 10-100 molar excess
Application Notes
This recombinant antigen is only intended to be used as a blocking agent to confirm antibody specificity with the corresponding antibody, catalog number NBP2-54690.

It is purified by IMAC chromatography, and the expected concentration is greater than 0.5 mg/ml.

For current lot information, including availability, please contact our technical support team click nb-technical@bio-techne.com

Theoretical MW
21 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Packaging, Storage & Formulations

Storage
Store at -20C. Avoid freeze-thaw cycles.
Buffer
PBS and 1M Urea, pH 7.4.
Preservative
No Preservative
Purity
>80% by SDS-PAGE and Coomassie blue staining

Alternate Names for beta-Actin Recombinant Protein Antigen

  • ACTB
  • actin, beta
  • actin, cytoplasmic 1
  • B-Actin
  • Beta Actin
  • beta cytoskeletal actin
  • betaActin
  • beta-Actin
  • BRWS1
  • I(2)-Actin
  • PS1TP5-binding protein 1
  • PS1TP5BP1

Background

Actin is a highly conserved protein involved in cell motility, structure, and integrity. Six functional actin isoforms exist and were named based on their electrophoretic mobility and distribution including alpha-cardiac muscle actin (aCAA), alpha-skeletal muscle actin (aSKA), alpha-smooth muscle actin (aSMA), beta-cytoplasmic actin (bCYA), gamma-cytoplasmic actin (gCYA) and gamma-smooth muscle actin (gSMA) (1). The cytoplasmic isoforms, beta-actin and gamma-actin, are nearly identical, differing by only 4 amino acids at the N-terminus with a theoretical molecular weight of 42 kDa (2). Beta-actin is ubiquitously expressed in eukaryotic cells and considered a housekeeping gene which is frequently used as a loading control for assays involving protein detection, such as Western blotting (3,4).

References

1. Vandekerckhove J, Weber K. 1978. At least six different actins are expressed in a higher mammal: an analysis based on the amino acid sequence of the amino-terminal tryptic peptide. J Mol Biol. 126(4):783-802. PMID: 745245

2. Gimona M, Vandekerckhove J, Goethals M, Herzog M, Lando Z, Small JV. (1994) Beta-actin specific monoclonal antibody. Cell Motil Cytoskeleton. 27(2):108-16. PMID: 8162619

3. Holden VI, Lenio S, Kuick R, Ramakrishnan SK, Shah YM, Bachman MA. (2014) Bacterial siderophores that evade or overwhelm Lipocalin 2 induce HIF-1a and pro-inflammatory cytokine secretion in cultured respiratory epithelial cells Infect Immun. 82(9):3826-36 PMID: 24980968

4. Tsai WL, Yeh PH, Tsai CY, Ting CT, Chiu YH, Tao MH, Li WC, Hung SC. (2016) Efficient Programming of Human Mesenchymal Stem Cell Derived Hepatocytes by Epigenetic Regulations. J. Gastroenterol. Hepatol. 32(1):261-269. PMID: 27218433

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Peptides and proteins are guaranteed for 3 months from date of receipt.

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FAQs for beta-Actin Recombinant Protein Antigen (NBP2-54690PEP). (Showing 1 - 5 of 5 FAQ).

  1. Why is beta actin used as a loading control in blotting ?
    • Beta actin is a highly-expressed protein found in all cells, and is considered a house keeping protein whose expression is necessary for any cell's proper functioning. For this reason it is used as a loading control to confirm that the same amount of total protein is loaded in each lane of an SDS-PAGE gel so that appropriate comparisons can be made between the actual proteins of interest in different samples.
  2. Are the beta-Actin antibodies validated in Simple Western?
    • Yes, we offer a few beta actin antibodies that have been tested in Simple Western: NBP1-47423, NB600-501, NB600-532, NB600-503, and NB100-56874.
  3. Do Beta-Actin antibodies come in lyophilized format?
    • ACTB antibodies like MAB8969 AND MAB8929 come in lyophilized form.
  4. What is the theoretical molecular weight for Beta-Actin antibodies?
    • The TMW for beta Actin antibodies is approximately 42 kDa.
  5. I wanted to know which of the two housekeeping genes B-actin or GAPDH can be used for best results. I intend to do an experiment to determine expression of IL-6 and TNF-alpha in liver and kidney tissues.
    • For homogenized tissue, beta-actin and GAPDH are both fine.

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Application Focus: I see an increase in LC3, now what?
 By Christina Towers, PhD.  Autophagy is highly conserved and tightly regulated process that all cell types use to recycle nutrients, particularly in the instance of stress1. As a result, even sm...  Read full blog post.

The use of Beta Actin (AC-15) as a loading control across multiple species
Actin is a fundamental component of the cytoskeleton, where it has the ability to create and break down actin filament formation in response to various cell needs.  Actin has six highly conserved isoforms, however beta and gamma actin are the two...  Read full blog post.

Tips on choosing an ideal loading control antibody for Western Blotting
Western blotting is one of the most commonly used antibody assay techniques in cell and molecular biology research since its development over three decades ago, and is considered the gold standard for protein detection and quantification. When p...  Read full blog post.

Beta-Actin's Role in Neuronal Plasticity
Beta-Actin is a highly conserved protein involved in cell growth, cytoskeletal and extracellular support structures and cell migration. Because beta-Actin is ubiquitously expressed in all eukaryotic cells, it is frequently used as a loading contro...  Read full blog post.

ACTB - an abundant cytoskeletal component with applications for gene expression analysis
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Bioinformatics

Gene Symbol ACTB