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Key Product Details

Species Reactivity

Validated:

Human, Mouse, Rat, Porcine, Bovine, Canine, Chicken, Drosophila (Negative), Feline, Fish, Guinea Pig, Hamster, Leech, Mammal, Primate, Rabbit, Sheep, Squirrel, Turkey, Zebrafish

Cited:

Human, Mouse

Applications

Validated:

Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Frozen, Knockdown Validated, Knockout Validated, Microarray, Simple Western, Western Blot

Cited:

WB

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 Clone # AC-15

Format

BSA Free

Concentration

1.0 mg/ml

Product Specifications

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.

Reactivity Notes

The antibody reacts with b-actin-expressing cells in bovine, canine, chicken, feline, carp, leech, guinea pig, human, mouse, porcine, rabbit, rat, and sheep. Does not react with amoeba or drosophila. Does not cross react with adult cardiac or skeletal muscle or amoeba beta actin. Not yet tested in other species. 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.

Specificity

Beta-actin (AC-15). 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.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1

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.

Description

This is the purified version of NB600-501.

Scientific Data Images for beta-Actin Antibody (AC-15) - BSA Free

Western Blot: beta-Actin Antibody (AC-15) - Purified [NB110-67828] - Whole cell extract of human fibroblasts was separated on SDS-PAGE and blotted with Monoclonal Anti-beta-Actin. The antibody was developed with Goat Anti-Mouse IgG, Peroxidase conjugate and AEC substrate. Lanes A: Antibody dilution 1:5,000 B: Negative control (only secondary antibody).
Immunocytochemistry/Immunofluorescence: beta-Actin Antibody (AC-15) - Purified [NB110-67828] - Staining in HS-68 cells.
Immunohistochemistry: beta-Actin Antibody (AC-15) - Purified [NB110-67828] - Staining in breast, Epithelium 40X.

Applications for beta-Actin Antibody (AC-15) - BSA Free

Application
Recommended Usage

Immunocytochemistry/ Immunofluorescence

10-40 ug/ml

Immunohistochemistry

1:10-1:500

Immunohistochemistry-Frozen

1:10-1:500

Western Blot

0.5-1 ug/ml
Application Notes
See Simple Western Antibody Database for Simple Western validation: tested in HeLa lysate; antibody dilution of 1:50; separated by size; detects a band at 54 kDa

Formulation, Preparation, and Storage

Purification

Protein A or G purified

Formulation

10mM PBS (pH 7.4)

Format

BSA Free

Preservative

0.9% Sodium Azide

Concentration

1.0 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: beta-Actin

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

Alternate Names

ACTB, betaActin

Entrez Gene IDs

60 (Human); 11461 (Mouse); 81822 (Rat)

Gene Symbol

ACTB

Additional beta-Actin Products

Product Documents for beta-Actin Antibody (AC-15) - BSA Free

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Product Specific Notices for beta-Actin Antibody (AC-15) - BSA Free

This is the purified version of NB600-501.

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.

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