beta-Actin Antibody (4C4)

Western Blot: beta-Actin Antibody (4C4) [NBP3-16886] - Analysis of extracts of various cell lines, using beta-Actin antibody (NBP3-16886) at 1:50000 dilution. Secondary antibody: HRP Goat Anti-Rabbit IgG (H+L) at 1:10000 dilution. Lysates/proteins: 25ug per lane. Blocking buffer: 3% nonfat dry milk in TBST. Detection: ECL Basic Kit. Exposure time: 60s.

Western Blot: beta-Actin Antibody (4C4) [NBP3-16886] - Analysis of extracts of various cell lines, using beta-Actin antibody (NBP3-16886) at 1:50000 dilution. Secondary antibody: HRP Goat Anti-Rabbit IgG (H+L) at 1:10000 dilution. Lysates/proteins: 25ug per lane. Blocking buffer: 3% nonfat dry milk in TBST. Detection: ECL Basic Kit. Exposure time: 60s.

Western Blot: beta-Actin Antibody (4C4) [NBP3-16886] - Analysis of extracts of HeLa cells, using beta-Actin antibody (NBP3-16886) at 1:10000-1:320000 dilution. Secondary antibody: HRP Goat Anti-Rabbit IgG (H+L) at 1:10000 dilution. Lysates/proteins: 25ug per lane. Blocking buffer: 3% nonfat dry milk in TBST. Detection: ECL Basic Kit. Exposure time: 180s.

Immunohistochemistry-Paraffin: beta-Actin Antibody (4C4) [NBP3-16886] - Human liver cancer tissue using beta -Actin Rabbit mAb at a dilution of 1:400 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate Buffer (pH 6.0) prior to IHC staining.

Western Blot: beta-Actin Antibody (4C4) [beta-Actin] - Western blot analysis of various lysates using beta-Actin Rabbit mAb at 1:50000 dilution incubated overnight at 4C.Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) at 1:10000 dilution.Lysates/proteins: 25 ug per lane.Blocking buffer: 3% nonfat dry milk in TBST.Detection: ECL Basic Kit .Exposure time: 10s.

• Reactivity: Hu, Mu, Rt, Po, Ch, Ze
• Applications: WB, ELISA, IHC
• Clone: 4C4
• Clonality: Monoclonal
• Host: Rabbit
• Conjugate: Unconjugated

beta-Actin Antibody (4C4) Summary

• Additional Information: Recombinant Monoclonal Antibody
• Immunogen: Recombinant protein of human beta-Actin.
• Isotype: IgG
• Clonality: Monoclonal
• Host: Rabbit
• Gene: ACTB
• Purity: Affinity purified

Applications/Dilutions

• Dilutions:
  • ELISA Recommended starting concentration is 1 μg/mL. Please optimize the concentration based on your specific assay requirements.
  • Immunohistochemistry 1:500 - 1:5000
  • Immunohistochemistry-Paraffin 1:500 - 1:5000
  • Western Blot 1:10000 -1:100000
• Theoretical MW: 42 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 (pH 7.3), 50% glycerol, 0.05% BSA
• Preservative: 0.09% Sodium Azide
• Purity: Affinity purified

Alternate Names for beta-Actin Antibody (4C4)

• 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.

FAQs for beta-Actin Antibody (NBP3-16886). (Showing 1 - 5 of 5 FAQ).

1. Why is beta actin used as a loading control in blotting ?  ..show answer..
   • 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?  ..show answer..
   • 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?  ..show answer..
   • ACTB antibodies like MAB8969 AND MAB8929 come in lyophilized form.
4. What is the theoretical molecular weight for Beta-Actin antibodies?  ..show answer..
   • 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.  ..show answer..
   • For homogenized tissue, beta-actin and GAPDH are both fine.

Bioinformatics

• Gene Symbol: ACTB