Transforming growth factor-beta 2 (TGF-beta 2) is a secreted protein known as a cytokine that performs many cellular functions and has a vital role during embryonic development. This gene is mapped to 1q41. It is an extracellular glycosylated protein. It is known to suppress the effects of interleukin dependent T-cell tumors. TGF-beta 2 is present at elevated levels in the aqueous humor of patients with primary open angle glaucoma(POAG). Studies have shown that TGF-beta 2 influences cultured trabecular meshwork cells, and it reduced outflow facility when perfused into cultured human anterior segments. In POAG, elevated expression of Gremlin by TM cells inhibited BMP4 antagonism of TGF-beta 2 and led to increased extracellular matrix deposition and elevated IOP. Transforming Growth Factor-beta 2 (TGF-beta 2), BioAssay(TM) ELISA Kit is a sandwich ELISA for quantitative detection of mouse TGF-beta 2 in cell culture supernates, serum and plasma (heparin, EDTA). Assay Range: 31.2-2000pg/ml Sensitivity: <10pg/ml Specificity: Natural and recombinant mouse TGF-beta 2 Crossreactivity: There is no detectable cross-reactivity with other relevant proteins Test Principle: This mouse TGF-beta 2 BioAssay(TM) ELISA Kit is a solid phase immunoassay specially designed to measure mouse TGF-beta 2 with a 96-well strip plate that is pre-coated with antibody specific for TGF-beta 2. The detection antibody is a biotinylated antibody specific for TGF-beta 2. The capture antibody is monoclonal antibody from rat, the detection antibody is polyclonal antibody from goat. The kit contains recombinant mouse TGF-beta 2 with immunogen: Expression system for standard: NSO, Immunogen sequence: A303-S414. The kit is analytically validated with ready to use reagents. To measure mouse TGF-beta 2, add standards and samples to the wells, then add the biotinylated detection antibody. Wash the wells with PBS or TBS buffer, and add Avidin-Biotin-Peroxidase Complex (ABC-HRP). Wash away the unbounded ABC-HRP with PBS or TBS buffer and add TMB. TMB is substrate to HRP and will be catalyzed to produce a blue color product, which changes into yellow after adding acidic stop solution. The density of the yellow product is linearly propotional to mouse TGF-beta 2 in the sample. Read the density of the yellow product in each well using a plate reader, and benchmark the sample wells readings against the standard curve to determine the concentration of mouse TGF-beta 2 in the sample. Kit Components: 353099A: Microtiter Strips, 1x96 wells 353099B: TGF-beta 2 Standard, 2x10ng 353099C: Anti-mouse TGF-beta 2 (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 all 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 minutes or at RT for 2 hours. Do not wash. 2. Add 100ul biotinylated antibody and incubate the plate at 37C for 60 minutes or at RT for 90 minutes. Wash plate 3 times with 0.01M PBS or TBS. 3. Add 100ul of ABC working solution and incubate the plate at 37C for 30 minutes or at RT for 40 minutes. Wash plate 5 times with 0.01M PBS or TBS. 4. Add 90ul of TMB color developing agent and incubate the plate at 37C in dark for 25-30 minutes or at RT for 30 minutes. 5. Add 100ul TMB Stop Solution and read.
