beta-Actin Antibody (AC-15) [HRP]

Western Blot: beta-Actin Antibody (AC-15) [HRP] [NB600-501H] - Analysis of beta-actin in NIH/3T3 and HEK293T cells using beta-actin antibody conjugated with HRP. Image from verified customer review.

Simple Western: beta-Actin Antibody (AC-15) [HRP] [NB600-501H] - Simple Western lane view shows a specific band for Beta-Actin in 0.2 mg/ml of Hela, MCF-7, SH-SY5Y, & Jurkat lysate(s). This experiment was performed under reducing conditions using the 12 - 230 kDa separation system.

Simple Western: beta-Actin Antibody (AC-15) [HRP] [NB600-501H] - Electropherogram image(s) of corresponding Simple Western lane view. Beta-Actin antibody was used at 1:500 dilution on Hela, MCF-7, SH-SY5Y, & Jurkat lysate(s).

Simple Western: beta-Actin Antibody (AC-15) [HRP] [NB600-501H] - Electropherogram image(s) of corresponding Simple Western lane view. Beta-Actin antibody was used at 1:100 dilution on Hela, MCF-7, SH-SY5Y, & Jurkat lysate(s).

Product Discontinued

beta-Actin Antibody (AC-15) [HRP] Summary

• Immunogen: This beta-Actin Antibody (AC-15) was made to a slightly modified Beta-cytoplasmic actin N-terminal peptide, Ac-Asp-Asp-Asp-Ile-Ala-Ala-Leu-Val-Ile-Asp-Asn-Gly-Ser-Gly-Lys, conjugated to KLH.
• Epitope: N-terminus of the beta isoform of actin.
• Specificity: In staining of chicken gizzard ultrathin tissue cryosections, the antibody labels the dense bodies and longitudinal channels linking consecutive dense bodies that are also occupied by desmin and the membrane-associated dense plaque. It does not stain adult cardiac and skeletal muscles except for traces due to contaminations of the sample with non-muscle cells, or if embryonic tissue is being used. The epitope recognized by the antibody is resistant to formalin-fixing and paraffin-embedding.
• Isotype: IgG1
• Clonality: Monoclonal
• Host: Mouse
• Gene: ACTB
• Purity: Protein G purified

Applications/Dilutions

• Dilutions:
  • ELISA
  • Immunohistochemistry
  • Immunohistochemistry-Frozen
  • Immunohistochemistry-Paraffin
  • Knockdown Validated
  • Knockout Validated
  • Simple Western
  • Western Blot
• 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.

Reactivity Notes

The antibody cross reacts with beta-actin expressing cells in carp, leech tissues (Hirudo medicinalis), ground squirrel. Does not cross react with adult cardiac, skeletal muscle, drosophila or amoeba beta actin. Mammal reactivity and Hamster reactivity reported in scientific literature (PMID: 25130694 and 24478435 respectively). Please note that this antibody is reactive to Mouse and derived from the same host, Mouse. Additional Mouse on Mouse blocking steps may be required for IHC and ICC experiments. Please contact Technical Support for more information.

Packaging, Storage & Formulations

• Storage: Store at 4C in the dark.
• Buffer: PBS
• Preservative: No Preservative
• Purity: Protein G purified

Notes

This conjugate is made on demand. Actual recovery may vary from the stated volume of this product. The volume will be greater than or equal to the unit size stated on the datasheet.

Alternate Names for beta-Actin Antibody (AC-15) [HRP]

• 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 (NB600-501H)(9)

Publications using NB600-501H

• Katie M Hamel, Connor T King, Maryn B Cavalier, Kara Q Liimatta, Grace L Rozanski, Timothy A King, Meggie Lam, Grace C Bingham, C Ethan Byrne, Diensn Xing, Bridgette M Collins-Burow, Matthew E Burow, Jorge A Belgodere, Melyssa R Bratton, Bruce A Bunnell, Elizabeth C Martin Breast Cancer-Stromal Interactions: Adipose-Derived Stromal/Stem Cell Age and Cancer Subtype Mediated Remodeling. Stem Cells Dev 2022-10-13 [PMID: 35579936] (Simple Western)
• Trent B, Liang Y, Xing Y et al. Polarized Lung Inflammation and Tie2/Angiopoietin-Mediated Endothelial Dysfunction during Severe Orientia tsutsugamushi Infection BioRxiv 2019-08-02 [PMID: 32119672] (WB, Mouse, Human)
• Davis SM, Collier LA, Foran EA et al. Neuroprotective activity of leukemia inhibitory factor is relayed through myeloid zinc finger-1 in a rat model of stroke Metab Brain Dis. 2019-04-15 [PMID: 30612292] (WB, Rat)
  
  Details:
  Citation using the HRP form of this antibody.
• Wu L, Zhang Z, Pan X et al. Expression and contribution of the HIF-1 alpha/VEGF signaling pathway to luteal development and function in pregnant rats Mol Med Rep 2015-09-02 [PMID: 26323652] (Rat)
• Pang X, Wang Z, Yin D, Zhang Z. Overexpression of hypoxia-inducible factor prolyl- hydoxylase attenuated by HCG-induced vascular endothelial growth factor expression in luteal cells. Mol Med Rep 2015-05-16 [PMID: 25975603]
• Vande Walle Lieselotte, Van Opdenbosch Nina, Jacques Peggy et al. Negative regulation of the NLRP3 inflammasome by A20 protects against arthritis. Nature 2014-08-07 [PMID: 25043000] (WB, Mouse)
  
  Details:
  Using the HRP conjugated version of NB600-501, catalog number NB600-501H.
• Brugger D, Buffler M, Windisch W. Development of an experimental model to assess the bioavailability of zinc in practical piglet diets. Arch Anim Nutr 2014-04-01 [PMID: 24646148] (WB, Porcine)
  
  Details:
  Using the HRP conjugated version of NB600-501, catalog number NB600-501H.
• Van Opdenbosch Nina, Gurung Prajwal, Vande Walle Lieselotte et al. Activation of the NLRP1b inflammasome independently of ASC-mediated caspase-1 autoproteolysis and speck formation. Nat Commun 2014-01-01 [PMID: 24492532] (WB)
  
  Details:
  Using the HRP conjugated version of NB600-501, catalog number NB600-501H.
• Liu L, Martin R, Kohler G, Chan C. Palmitate induces transcriptional regulation of BACE1 and presenilin by STAT3 in neurons mediated by astrocytes. Exp Neurol 2013-10-01 [PMID: 23968646] (WB, Rat)
  
  Details:
  Using the HRP conjugated version of NB600-501, catalog number NB600-501H.

Reviews for beta-Actin Antibody (NB600-501H) (3)

Average Rating: 5

(Based on 3 reviews)

We have 3 reviews tested in 3 species: Human, Mouse, Human and Mouse.

reviewed by: Verified Customer Simple Western Mouse 04/23/2019

Summary

• Application: Simple Western
• Sample Tested: Mouse skeletal muscle
• Species: Mouse
• Lot: 087M-4860V-112718-H

reviewed by: Fernanda gutierrez rodrigues WB Human 09/08/2017

Summary

• Application: Western Blot
• Sample Tested: 293T lysate
• Species: Human
• Lot: 037M4782V-062817-H

reviewed by: Verified Customer WB Human and Mouse 11/02/2016

Summary

• Application: Western Blot
• Sample Tested: whole cell lysate prepared with Cell Signal Lysis buffer
• Species: Human and Mouse
• Lot: 065M4837V-092016-H

FAQs for beta-Actin Antibody (NB600-501H). (Showing 1 - 6 of 6 FAQs).

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.
6. What is the immunogen sequence of this Beta-Actin antibody?
   • A sequence (Ac-Asp-Asp-Asp-Ile-Ala-Ala-Leu-Val-Ile-Asp-Asn-Gly-Ser-Gly-Lys) within the N-terminal region of beta Actin, conjugated to KLH.

Bioinformatics

• Gene Symbol: ACTB