Interleukin-8, also called neutrophil-activating peptide-1 or SCYB8, is a tissue-derived peptide secreted by several types of cells in response to inflammatory stimuli. Monocyte-derived neutrophil chemotactic factor (MDNCF/IL-8, suggested gene symbol IL8) is a cytokine that chemoattracts and activates neutrophils (1). IL-8 is produced and released from human adipose tissue and from isolated adipocytes in vitro, which may indicate that IL-8 from adipose tissue could be involved in some of the obesity-related complications (2). The MDNCF/IL-8 gene is placed on the human gene map at position 4q12-q21. This is the same location where at least three other members (platelet factor 4, melanoma growth stimulatory activity, and interferon-gamma induced factor) of the platelet factor 4 gene superfamily reside (1). Human IL-8 consists of 99 amino acids in precursor form and 79 amino acids in mature form. Intended Use: IL-8 BioAssay(TM) ELISA Kit, Human is a sandwich ELISA for quantitative detection of human IL-8 in cell culture supernates, serum and plasma (heparin, EDTA, citrate). Range: 15.6-1000pg/ml (cell culture supernates) 7.8-500pg/ml (serum, plasma) Sensitivity: < 1pg/ml Specificity: Natural and recombinant human IL-8 Crossreactivity: No detectable crossreactivity with other relevant proteins Kit Components: 143935A: Microtiter Strips, 1x96 wells 143935B: IL-8 Standard, 2x10ng 143935C: Anti-human IL-8 (Biotin), 1x130ul USB Cat No Kit Component 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 Assay Summary: 1. Add samples and standards and incubate the plate at 37C for 90 minutes. Do not wash. 2. Add biotinylated antibodies and incubate the plate at 37C for 60 minutes. Wash plate 3 times with 0.01M TBS. 3. Add ABC working solution and incubate the plate at 37C for 30 minutes. Wash plate 5times with 0.01M TBS. 4. Add TMB color developing agent and incubate the plate at 37C in dark for 25-30 minutes. 5. Add TMB stop solution and read.. Storage and Stability: Store *143935B 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.
