EGF-containing fibulin-like extracellular matrix protein 1 is a protein that in humans is encoded by the EFEMP1 gene. This gene encodes a member of the fibulin family of extracellular matrix glycoproteins. Like all members of this family, the encoded protein contains tandemly repeated epidermal growth factor-like repeats followed by a C-terminal fibulin-type domain. This gene is upregulated in malignant gliomas and may play a role in the aggressive nature of these tumors. Mutations in this gene are associated with Doyne honeycomb retinal dystrophy. Alternatively spliced transcript variants that encode the same protein have been described. The Fibulin-3/EFEMP1 BioAssay(TM) ELISA Kit is a Sandwich High Sensitivity ELISA for quantitative detection of human Fibulin-3/EFEMP1 in cell culture supernates, cell lysates, serum, plasma (Heparin or EDTA). Detection Range: 312pg/ml-20,000pg/ml Sensitivity: <10pg/ml Specificity: Recognizes natural and recombinant Human Fibulin-3/EFEMP1. There is no detectable cross-reactivity with other relevant proteins. Test Principle: This Fibulin-3/EFEMP1 BioAssay(TM) ELISA Kit is a Sandwich High Sensitivity ELISA is a solid phase immunoassay specially designed to measure human Fibulin-3/EFEMP1 with a 96-well strip plate that is pre-coated with antibody specific for Fibulin-3/EFEMP1. The detection antibody is a biotinylated antibody specific for Fibulin-3/EFEMP1. The kit contains recombinant human Fibulin-3/EFEMP1 with immunogen: Expression system for standard: CHO, Immunogen sequence: Q18-F493. To measure human Fibulin-3/EFEMP1, add standards and samples to the wells, then add the biotinylated detection antibody. Wash the wells with PBS buffer, and add Avidin-Biotin-Peroxidase Complex (ABC-HRP). Wash away the unbounded ABC-HRP with PBS 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 proportional to human Fibulin-3/EFEMP1 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 human Fibulin-3/EFEMP1 in the sample. Kit Components: 396663A: Microtiter Strips, 1x96 wells *396663B: Fibulin-3/EFEMP1 Standard, 2x20ng 396663C: Anti-human Fibulin-3/EFEMP1 (Biotin), 1x100ul (1:100 dilution) 143696: Avidin-Biotin-Peroxidase Complex (ABC), 1x100ul (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 143702: Wash Buffer, 25X, 1x20ml Storage and Stability: Store *396663B 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 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 Working Wash Buffer. 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 Working Wash Buffer. 4. Add 90ul of TMB color developing agent and incubate the plate at 37C in dark for 15-25 minutes or at RT for 30 minutes. 5. Add 100ul TMB Stop Solution and read.
