Anti-Human/Mouse/Rat Activin A

Product Datasheet

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Anti-Human Activin A

Purified No Carrier Protein

Monoclonal Antibody

Prod. No.:	A374
Clone:	69403
Isotype:	Mouse IgG₁
Pkg. Size:	1 mg, 500 µg
Storage:	-20°C to -70°C Detailed storage instructions below.

Description

Specificity:

Mouse Anti-Human Activin A (Clone 69403) recognizes an epitope on Human Activin A. This monoclonal antibody was purified using multi-step affinity chromatography methods such as Protein A or G depending on the species and isotype.

Background:

Activin A is a member of the TGF beta super family. It was originally identified and isolated because it stimulates the release of follicle-stimulating hormone (FSH). Activins are structurally related to inhibins, they share common beta subunits, but they exhibit opposite biological activities. Activins are homodimers of beta subunits and inhibins are heterodimers of an alpha subunit and a beta subunit. Since the initial discovery of Activin A, it has been shown to regulate cell differentiation, nerve cell survival, bone growth promotion, cell proliferation, and apoptosis. Activin A expression has been detected in prostate cancer, colon cancer and breast cancer. Because Activin A is able to inhibit cell proliferation by modulating the Rb pathway, it is thought to inhibit cancer development. This may not be the only mechanism by which activin may play a preventative role in cancer development. Activin has also been shown to increase the expression of a protein whose decreased expression is correlated with increased metastatic capacity. In addition, activin has an inhibitory effect on endothelial cell growth and can thereby block angiogenesis, preventing tumor growth and metastasis.

The activin/inhibin nomenclature reflects the subunit composition of the proteins: activin A (βA - βA), activin B (βB - βB), activin AB (βA - βB), inhibin A (α - βA) and inhibin B (α - βB). At present, little is known about the contribution of the other β subunits to activin or inhibin formation and biology. At the amino acid sequence level, the mature human βA subunit is 100% identical to mouse βA, while the human and mouse α subunits share approximately 80% identity. Similarly to other TGF-β family members, activins exert their biological activities through binding to the heterodimeric complex composed of two membrane spanning serine-threonine kinases designated type I and type II. Two forms of activin receptor type I (Act RI-A and Act RI-B) and two forms of activin receptor type II (Act RII-A and Act RII-B) have been identified. Activin binds directly to Act RII, the complex then associates with Act RI and initiates signaling.

Known Species Cross Reactivity: Ms+, Hu+, Rt+

Host Species

Mouse

Immunogen

Purified Recombinant Human Activin A (>98%)

Formulation

This monoclonal antibody has been 0.2 µm filtered and lyophilized from modified Dulbecco's phosphate buffered saline (1X PBS) pH 7.2 - 7.3 containing 5.0% w/v trehalose with no calcium, magnesium or preservatives present.

Reconstitution

This lyophilized antibody may be aseptically reconstituted with sterile PBS to a final volume of 1.0 ml. This will result in a final antibody concentration of 500 µg/ml.

Endotoxin

<0.1 EU/µg as determined by the LAL method

Storage and Stability

The lyophilized antibody can be stored desiccated at -20°C to -70°C for up to twelve months. The reconstituted antibody can be stored for at least four weeks at 2-8°C. For long-term storage of the reconstituted antibody, aseptically aliquot into working volumes and store at -20°C to -70°C in a manual defrost freezer. Avoid repeated freeze thaw cycles. No detectable loss of activity was observed after six months.

Reported Applications

ELISA Cap, IHC (NBF/Par.), N, WB

ELISA Sandwich: This antibody is useful as the capture antibody in a sandwich ELISA. The suggested coating concentration is 2-8 µg/ml.

IHC (NBF/Par.): This antibody should give satisfactory staining results when used at a concentration of 25 µg/ml. The recommended secondary antibody for IHC is Goat Anti-Mouse IgG (H&L)-Horseradish Peroxidase (Leinco Prod. No.: M114). For chromogenic detection with high signal and low background use 3, 3'-Diaminobenzidine Tetrahydrochloride (DAB) Substrate Kit (Leinco Prod. No.: D100) or Optional Metal Enhanced DAB Substrate Kit (Leinco Prod. No.: K107).

Neutralization: This antibody is useful for neutralization of Human/Mouse/Rat Activin A bioactivity. The antibody dose required to neutralize 50% (ND₅₀) of the biological activity of Human/Mouse/Rat Activin A (at 0.75 ng/ml) is 0.02 - 0.06 µg/ml.

Western Blotting: To detect Human/Mouse/Rat Activin A this monoclonal antibody can be used at a concentration of 1-2 µg/ml. This monoclonal antibody should be used in conjunction with compatible second-step reagents such as Goat Anti-Mouse IgG (H&L)-Horseradish Peroxidase (Leinco Prod. No.: M114) and a chromogenic substrate such as TMB HRPO Membrane/IHC Substrate UltraSensitive™ One Component "Ready Use" (Leinco Prod. No.: T343). The detection limit for Human/Mouse/Rat Activin A is 5 ng/lane under either reducing or non-reducing conditions. The sensitivity of detection may increase up to 50 fold when a chemiluminescent substrate is used.

Each investigator should determine their own optimal working dilution for specific applications.

Country of Origin

USA