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.

Do Beta-Actin antibodies come in lyophilized format?

ACTB antibodies like MAB8969 AND MAB8929 come in lyophilized form.

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.

What is the theoretical molecular weight for Beta-Actin antibodies?

The TMW for beta Actin antibodies is approximately 42 kDa.

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.

Mouse beta-Actin ELISA Kit (Colorimetric)

Name: Mouse beta-Actin ELISA Kit (Colorimetric)
Brand: Novus Biologicals
SKU: NBP3-57006

Novus Biologicals, part of Bio-Techne | Catalog # NBP3-57006

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Product Specifications
Precision
Kit Contents
Preparation & Storage
Background
Product Documents
Specific Notices

Key Product Details

Sample Type & Volume Required Per Well

Tissue homogenates, cell lysates, cell culture supernatants and other biological fluids (100 uL)

Sensitivity

0.69 ng/mL (example only; lot dependent)

Assay Range

1.56 - 100 ng/mL (example only; lot dependent)

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Product Specifications

Assay Type

Sandwich ELISA

Kit Type

ELISA Kit (Colorimetric)

Reactivity

Mouse

Description

Assay Length: 4.5 hours

Precision

Intra-Assay Precision (Precision within an assay) %CV < 10 (example only; lot dependent)

Inter-Assay Precision (Precision between assays) %CV < 12 (example only; lot dependent)

Kit Contents for Mouse beta-Actin ELISA Kit (Colorimetric)

Pre-coated 96T strip plate, Plate sealer for 96 wells, Standard, Diluent Buffer, Detection Reagent A, Detection Reagent B, TMB Substrate, Stop Solution, Wash Buffer (30 x concentrate), Instruction manual

Preparation and Storage

Stability & Storage

Storage of components varies. See protocol for specific instructions.

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

Gene Symbol

ACTB

Additional beta-Actin Products

Product Documents for Mouse beta-Actin ELISA Kit (Colorimetric)

Product Datasheets

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COA

Product Specific Notices for Mouse beta-Actin ELISA Kit (Colorimetric)

This product is for research use only and is not approved for use in humans or in clinical diagnosis. ELISA Kits are guaranteed for 6 months from date of receipt.

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