| Neutrophil Gelatinase-Associated Lipocalin, BioAssay(TM) ELISA Kit is a sandwich ELISA intended for the quantitative determination of NGAL in human serum, plasma, tissue homogenates and urine. Detection Range: 15.6-1000pg/ml Intra-assay Precision: %CV <8 Inter-assay Precision: %CV <10 Sensitivity: The minimum detectable dose of human NGAL is typically less than 3.9pg/ml. The sensitivity of this assay, or Lower Limit of Detection (LLD) was defined as the lowest protein concentration that could be differentiated from zero. It was determined the mean O.D value of 20 replicates of the zero standard added by their three standard deviations. Specificity: This assay has high sensitivity and excellent specificity for detection of human NGAL. No significant crossreactivity or interference between human NGAL and analogs was observed. Note: Limited by current skills and knowledge, it is impossible for us to complete the crossreactivity detection between human NGAL and all the analogs. Therefore, crossreaction may still exist. Kit Components: 227841A: Microtiter Strips, 1x48 wells or 1x96 wells. Pre-coated, ready to use. 227841B: Standard, 2x1vial 227841C: NGAL (Biotin), 100X, 1x60ul or 1x120ul 227841D: Avidin (HRP), 100X, 1x60ul or 1x120ul 227841E: Biotin Diluent, 1x7.5ml or 1x15ml 227841F: Avidin (HRP) Diluent, 1x7.5ml or 1x15ml 227841G: Sample Diluent, 1x25ml or 1x50ml 227841H: Wash Buffer, 25X, 1x10ml or 1x20ml 227841J: TMB Substrate, 1x5ml or 1x10ml 227841K: Stop Solution, 1x5ml or 1x10ml Note: The Stop Solution (227841K) provided with this kit is an acid solution. Wear eye, hand, face, and clothing protection when using this material. Storage and Stability: Store kit at 4C. The unopended kit is stable for 6 months after receipt. Once opened, the kit is stable up to 4 weeks at 4C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Assay Summary: 1. Prepare reagents, standards and samples as instructed. 2. Add 100ul standard or sample to each well. Incubate for 2 hours at 37C 3. Remove the liquid of each well do not wash. 4. Add 100ul NGAL (Biotin), 1X to each well. Incubate for 1 hour at 37C 5. Aspirate and wash 3 times. 6. Add 100ul Avidin (HRP), 1X to each well. Incubate for 1 hour at 37C 7. Aspirate and wash 5 times. 8. Add 90ul TMB substrate to each well Incubate for 15-30 minutes at 37C. Protect from light 9. Add 50ul Stop Solution to each well. Read at 450nm within 5 minutes. |
