beta-Actin Antibody (4C2)

Product Discontinued

beta-Actin Antibody (4C2) Summary

• Immunogen: Peptide comprising the N-terminal nonapeptide of beta-cytoplasmic actin with an acetylated N-terminus (Ac-DDDIAALVV-COOH) coupled to keyhole limpet hemocyanin through the cysteine residue.
• Specificity: The monoclonal antibody Anti-b-CYA, clone 4C2 is an antibody against beta cytoplasmic actin, the antibody is highly specific for beta-cytoplasmic actin and does not cross react with other actin isoforms.
• Isotype: IgG1
• Clonality: Monoclonal
• Host: Mouse
• Gene: ACTB
• Purity: Protein A or G purified

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry/ Immunofluorescence
  • Immunohistochemistry
  • Immunohistochemistry-Frozen
  • Immunohistochemistry-Paraffin 1:100 - 1:500
  • Western Blot 1:1000 - 1:5000
• Application Notes: Optimal antibody dilution should be determined by titration; recommended range is 1:100 - 1:500 for Immunohistochemistry with avidin-biotinylated horseradish peroxidase complex (ABC) as detection reagent..

Reactivity Notes

Human, Porcine, Rabbit and Rat. The epitope recognized by Clone 4C2 is highly conserved and the antibody is therefore expected to react with amny other species.

Packaging, Storage & Formulations

• Storage: Store at 4C. Do not freeze.
• Buffer: PBS
• Preservative: 0.09% Sodium Azide
• Concentration: 1 mg/ml
• Purity: Protein A or G purified

Alternate Names for beta-Actin Antibody (4C2)

• 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. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for beta-Actin Antibody (NBP1-97721)(1)

Publication using NBP1-97721

• Tomasek JJ, Haaksma CJ, Schwartz RJ, Howard EW. Whole animal knockout of smooth muscle alpha-actin does not alter excisional wound healing or the fibroblast-to-myofibroblast transition Wound Repair Regen 2013-01-01 [PMID: 23253249] (WB, Mouse)

FAQs for beta-Actin Antibody (NBP1-97721). (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.

Bioinformatics

• Gene Symbol: ACTB
• Entrez:
  • 60(Human)
  • 11461(Mouse)
  • 81822(Rat)