Beta Actin ELISA Kit (ACTB)

Full Name: Beta Actin ELISA Kit (ACTB)
Reactivity: Human
Sample Type: Tissue Homogenates, Biological Fluids, Cell Lysates
Sensitivity: 0.188 ng/ml

INTRODUCTION

One of the two cytoskeletal actins found outside of muscles is beta-actin (ACTB). Due to its high conservation, it serves as an endogenous housekeeping gene to normalise molecular expression investigations. However, recent studies show its expression can change in response to biochemical stimuli, during growth and differentiation and in disease states. ACTB binds GTP and hydrolyses GTP into GDP which leads to a conformational change of association with other proteins by phosphorylation. The regulation of beta-actin dynamics has recently been studied as one mechanism by which cytoskeletal tension controls cell migration and migration velocity. The precise mechanism is unknown, however a study has demonstrated that regulated phosphorylation, depending on whether it occurs in rapid or slow developing cells, can result in either an increase or decrease in actin filament turnover.

Beta-actin is similar to tubulin in that it has two globular domains (G-actin) connected via a flexible, filamentous linker region. Actin polymers are often referred to as microfilaments or monofilaments. Actins are always paired, with one monomer being on the outside of the polymer while the other is on the inside where they interact with each other via their respective G-actin domains. This interaction maintains a polarity in actin filaments, so that they can attach to and move along other actin filaments. Monomers contain one or more globular G-actin domains which are situated at the 3′ end of the molecule. In each monomer, two G-actin domains are oriented toward each other to form a long rod-like structure called the “rod”, which is approximately 20 nm in diameter. There is no space between adjacent monomers because they are packed tightly together like radial multiples in a hexagonal close pack.

INTENDED USE

Human beta actin ELISA kit can detect levels of beta cytoskeletal actin (beta actin, ACTB, cytoplasmic 1, PS1TP5-binding protein) present is human cell lysates, tissue homogenate and other biological fluids.

CONTENT

All reagents supplied need to be stored at 2 °C – 8 °C, unopened reagents will retain reactivity until expiration date. Do not use reagents beyond this date.

One 96-Well Plate: Pre-coated with anti-beta-actin antibody.
Standards: Lyophilized recombinant.
Sample/Standard Dilution Buffer.
Biotinylated-labelled Antibody.
Antibody Dilution Buffer.
HRP-Streptavidin Conjugate (SABC).
SABC Dilution Buffer.
TMB Substrate.
Wash Buffer (25x).
Plate Sealer.
Product Instructions.

TYPICAL RESULTS

For this beta actin ELISA kit it is recommended that a standard curve is generated for each assay carried out.

Standard Curve: 0, 0.312, 0.625, 1.25, 2.5, 5, 10, 20 ng/ml.
Reactivity: Human
Sensitivity: 0.188 ng/ml
Range: 0.313 – 20 ng/ml
Principle: Sandwich
Application: Research Use Only.

ASSAY CHARACTERISTICS

– Specificity: Highly specific for beta-actin, no cross reactivity or interference between beta-actin and analogues was detected.
– Recovery: Cell Lysates (90 – 105%)
– Precison Intra-Assay: CV < 8%.
– Precison Inter-Assay: CV < 10%.
– Stability: Less than 10%.

REFERENCES

A pan-cancer analysis of the prognostic and immunological role of β-actin (ACTB) in human cancers. (2021) 12 (1): 6166-6185. Gu Y., et al.
Beta-actin specific monoclonal antibody. Cell Motil Cytoskeleton. (1994) 27 (2): 108-16. Gimona M., et al.
Beta-cytoplasmic actin localization in vertebrate glomerular podocytes. Arch Histol Cytol. (2009) 72 (3): 165-74. Ichimura K., et al.

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