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Streamline studies of diabetic nephropathy and other processes where measurement of βig-H3/TGFBI/BIGH3 levels delivers necessary insights.
FOR RESEARCH USE ONLY
For researchers monitoring renal dysfunction during the early stages of diabetes, the clinical progression of diabetic neuropathy, urinary system cancers, or other studies monitoring levels of βig-H3/TGFBI/BIGH3 our Exocell βig-H3 ELISA kits can provide accurate measurement of βig-H3 from urine, serum or supernatant samples of human or mouse origin. Both kits are configured as quantitative sandwich ELISAs that use biotinylated detection antibodies. Measurement occurs through streptavidin-conjugated horse radish peroxidase (HRP) which binds to the biotinylated antibody, and a colorimetric HRP substrate that absorbs at 450 nm. Color intensity in the βIG-H3 ELISAs is proportional to the amount of βig-H3 captured on the plate.
The βIG-H3 ELISA Kits are part of our Exocell portfolio and deliver the same reliable results and expert support that Exocell customers know to expect, including reagents that are ready-to-use either as is or with only moderate dilution. Quantitation is simplified through the use of the Creatinine Companion, which enables normalization to creatinine levels to account for variable urine concentration.
What’s a sandwich ELISA?
In a sandwich ELISA, antigen measurement occurs by building an antibody-antigen-antibody “sandwich.” First, antibody that recognizes the target antigen is immobilized in the wells of a multi-well plate. The sample is added to the plate and target antigen binds to the immobilized antibody. After washing, a solution containing soluble antibody that recognizes the target antigen is added to detect and measure the amount of target antigen present in the wells.
To make a sandwich ELISA quantitative, it is critical to achieve uniform and consistent coating of the initial antibody to the multi-well plate.
Why use a sandwich ELISA?
Sandwich ELISAs are useful when purified target antigen is not available to create a competitive ELISA plate or when antibody to the target antigen is plentiful.
What is direct vs. indirect detection?
Direct versus indirect detection refers to how HRP is introduced into the assay. With direct detection, HRP is conjugated directly to the antibody that recognizes the analyte. This is the simpler and quicker of the two methods as only a single antibody is needed. With indirect detection, a primary antibody recognizes the analyte, and then a secondary antibody conjugated to HRP recognizes the primary antibody. The addition of the secondary antibody adds wash steps, increasing the time and labor involved in the assay workflow as well as adding steps where variability can be introduced.
βig-H3 (beta-induced gene H3 ) is a 68 kD protein induced in diverse cell types by Transforming Growth Factor-β (TGF-β). βig-H3 is secreted into the extracellular matrix and is involved in cell growth, differentiation, adhesion, and wound healing and expression of the βig-H3 gene is altered in various conditions including tumorigenesis, breast cancer, corneal dystrophy, osteogenesis, and atherothrombosis. βig-H3 protein levels are elevated in the urine of patients with cyclosporine nephrotoxicity as well as type 2 diabetes, where it positively correlates with the albumin excretion rate, making it a useful biomarker of renal dysfunction during early-stage diabetes and for monitoring clinical progression of diabetic nephropathy.
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