| Activin A BioAssay(TM) ELISA Kit is a sandwich ELISA for quantitative detection of human Activin A in cell culture supernates, serum, plasma (heparin, EDTA) and saliva. Activin A is a homodimer of 14kD beta-A. Activin A, a cytokine member of the transforming growth factor-beta superfamily, is expressed locally by the mesenchymal component of the hemopoietic microenvironment. Its expression is regulated on the mRNA level by different cytokines, and the biological activity of the protein is tightly controlled by several inhibitory molecules. Inhibins and activins are members of the transforming growth factor beta superfamily and are known to modulate the growth and differentiation of several cell types. Inhibins and activins inhibit and activate, respectively, the secretion of follitropin by the pituitary gland. Inhibins/activins are involved in regulating a number of diverse functions such as hypothalamic and pituitary hormone secretion, gonadal hormone secretion, germ cell development and maturation, erythroid differentiation, insulin secretion, nerve cell survival, embryonic axial development or bone growth, depending on their subunit composition. Inhibins appear to oppose the functions of activins. Assay Range: 15.6pg/ml-1000pg/ml Sensitivity: <12pg/ml Specificity: Natural and recombinant Activin A Crossreactivity: There is no detectable cross-reactivity with other relevant proteins Test Principle: This human Activin A BioAssay(TM) ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for Activin A has been precoated onto 96-well plates. Standards (CHO cells, Immunogen Sequence G311-S426)) and test samples are added to the wells, a biotinylated detection monoclonal antibody from goat specific for Activin A is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the human Activin A amount of sample captured in plate. Kit Components: 143706A: Microtiter Strips, 1x96 wells *143706B: Activin A Standard, 2x10ng 143706C: Anti-human Activin A (Biotin), 1x130ul (1:100 dilution) 143696: Avidin-Biotin-Peroxidase Complex (ABC), 1x130ul (1:100 dilution) 143697: Sample Diluent Buffer, 1x30ml 143698: Antibody Diluent Buffer, 1x12ml 143699: ABC Diluent Buffer, 1x12ml 143700: TMB Color Developing Agent, 1x10ml 143701: TMB Stop Solution, 2N H2SO4, 1x10ml Storage and Stability: Store *143706B powder at 4C. Once reconstituted store at 4C for up to 12 hours or at -20C for up to 48 hours. Store other components at 4C. Stable for 6 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Assay Summary: 1. Add 100ul of samples and standards and incubate the plate at 37C for 90 min. Do not wash. 2. Add 100ul biotinylated antibody and incubate the plate at 37C for 60 min. Wash plate 3 times with 0.01M PBS or TBS. 3. Add ABC working solution and incubate the plate at 37C for 30 minutes. Wash plate 5 times with 0.01M PBS or TBS. 4. Add TMB color developing agent and incubate the plate at 37C in dark for 15-20 minutes. 5. Add 100ul TMB stop solution and read. |
