The Human Haptoglobin Related Protein (HPR) ELISA, ECL Kit is a sensitive chemiluminescent sandwich enzyme immunoassay for the in vitro quantitative measurement of HPR in human serum, plasma and other biological fluids.
Test Principle:The microtiter plate provided in this kit has been pre-coated with an antibody specific to HPR. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to HPR. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. The addition of substrates A and B mixture generates glow light emission kinetics. Upon plate development, the intensity of the emitted light is proportional to the HPR level in the sample or standard.
Detection Range:0.781-50ng/ml
Sensitivity:<0.25ng/ml
Precision:CV(%) = (SD/mean) X 100Intra-Assay: CV<10%Inter-Assay: CV<12%
Kit Components:*157711A: Microtiter Plate, 1x96 wells, Pre-coated with antibody; ready to use
*157711B: Standard, 2x1vial 157711C: Standard Diluent, 1x20ml*157711D: Detection Reagent A (green), 1x120ul *157711E: Detection Reagent B (red), 1x120ul 157711F: Assay Diluent A (2x), 1x12ml157711G: Assay Diluent B (2x), 1x12ml157711H: Substrate A, 1x10ml157711K: Substrate B, 1x2ml157711L: Wash Buffer (30x), 1x20ml
Storage and Stability:Store *157711A, *157711B, *157711D and *157711E at -20°C. Store all the other components at 4°C. Unused kit is stable for 6 months. Once kit components are opened, it is highly recommended to use remaining reagents within 1 month provided this is within the expiration date of the kit. For maximum recovery of product, centrifuge the original vials after thawing and prior to removing the cap.
Materials Required But Not Supplied:1. Luminometer capable of reading 96-well microplates with the following parameters: lag time 30.0secs; read time 1.0 sec/well . 2. Precision single or multi-channel pipettes and disposable tips
3. Eppendorf Tubes for diluting samples
4. Deionized or distilled water
5. Absorbent paper for blotting the microtiter plate
6. Container for Wash Solution
Sample Preparation and Storage:Serum:Use a serum separator and allow samples to clot for two hours at room temperature or overnight at 4°C before centrifugation for 20 minutes at approximately 1000xg. Assay freshly prepared serum immediately or store samples in aliquots at -20°C or -80°C for later use. Avoid repeated freeze/thaw cycles.
Plasma:Collect plasma using EDTA or heparin as anticoagulant. Centrifuge samples for 15 minutes at 1000xg at 2-8°C within 30 minutes of collection. Remove plasma and assay immediately or store samples in aliquots at -20°C or -80°C for later use. Avoid repeated freeze/thaw cycles.
Note: Serum/plasma samples require about 10,000-fold dilution. To prepare a 1:10,000-fold dilution of sample: Transfer 10ul sample to 990ul PBS; this yields a 1:100 dilution. Dilute further by transferring 10ul of the 1:10 diluted sample to 990ul PBS; this yields a 1:10,000 dilution. Mix thoroughly at each stage of the dilution process. Samples should be diluted using 0.02M PBS (pH 7.0-7.2).
Other Biological Fluids:Centrifuge samples for 20 minutes at 1000×g. Remove particulates and assay immediately or store samples in aliquots at -20°C or -80°C. Avoid repeated freeze/thaw cycles.
Assay Procedure Summary:1. Prepare all reagents, samples and standards
2. Add 100ul standard or sample to each well. Incubate 2 hours at 37°C
3. Aspirate and add 100ul prepared Detection Reagent A. Incubate 1 hour at 37°C
4. Aspirate and wash 3 times
5. Add 100ul prepared Detection Reagent B. Incubate 30 minutes at 37°C
6. Aspirate and wash 5 times
7. Add 100ul Substrate Solution. Incubate 10 minutes at 37°C
8. Read RLU immediately
仕様
Size：96Tests
Source Antigen：Human
Specificity：This sandwich assay has high sensitivity and excellent specificity for detection of Haptoglobin Related Protein (HPR). No significant cross-reactivity or interference between Haptoglobin Related Protein (HPR) and analogues was observed.
