Phosphoserine Phosphatase (PSPH) BioAssay™ ECL ELISA Kit (Human)

Phosphoserine Phosphatase (PSPH) BioAssay™ ECL Kit (Human) is a quantitative, in vitro, Chemiluminescence-based sandwich immunoassay (ECL) suitable for use in Tissue homogenates and other biological fluids.||Test Principle:|The microtiter plate provided in this kit has been pre-coated with an antibody specific to Phosphoserine Phosphatase. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Phosphoserine Phosphatase. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. Then the mixture of substrate A and B is added to generate glow light emission kinetics. Upon plate development, the intensity of the emitted light is proportional to the Phosphoserine Phosphatase (PSPH) level in the sample or standard.||Detection Range:|0.156-10ng/ml||Sensitivity:|The minimum detectable dose of this kit is typically less than 0.056ng/ml.||Specificity:|This sandwich assay has high sensitivity and excellent specificity for detection of Phosphoserine Phosphatase (PSPH). No significant cross-reactivity or interference between Phosphoserine Phosphatase (PSPH) and analogues was observed. ||Stability:|The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition. To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.||Precision:|Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level were tested 20 times on one plate, respectively. |Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level were tested on 3 different plates, 8 replicates in each plate. | CV(%) = SD/meanX100| Intra-Assay: CV<10%| Inter-Assay: CV<12%||Kit Components:|Microtiter Plate, 1x96 wells, Pre-coated, ready to use, well strip plate|Plate sealer for 96 wells|Standard, 2x1 vial|Standard Diluent, 1x20ml|Detection Reagent A, 1x120ul|Detection Reagent B, 1x120ul|Assay Diluent A, 1x12ml|Assay Diluent B, 1x12ml|Substrate A, 1x10ml|Substrate B, 1x2ml|Wash Buffer, 30X, 1x20ml||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 pipette tips with 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||Protocol 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 value immediately.||Sample Collection:|Serum:|Use a serum separator tube and allow samples to clot for two hours at RT or overnight at 4°C before centrifugation for 20 minutes at approximately 1000×g. Assay freshly prepared serum immediately or store samples in aliquot at -20°C or -80°C for later use. Avoid repeated freeze/thaw cycles.||Plasma:|Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples for 15 minutes at 1000×g 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.||Other Biological Fluids:|Centrifuge samples for 20 minutes at 1000xg. Remove particulates and assay immediately or store samples in aliquot at -20°C or -80°C for later use.|Avoid repeated freeze/thaw cycles.||Note:|Samples to be used within 5 days may be stored at 4°C, otherwise samples must be stored at -20°C ( 1 month) or -80°C ( 2 months) to avoid loss of bioactivity and contamination.|Sample hemolysis will influence the result, so hemolytic specimens cannot be detected.|When performing the assay, bring samples to room temperature.||Storage and Stability:|1. For unopened kit: All the reagents should be kept according to the labels on vials. The Standard, Detection Reagent A, Detection Reagent B and the 96-well strip plate should be stored at -20°C upon receipt while the others should be at 4°C.|2. For opened kit: When the kit is opened, the remaining reagents still need to be stored according to the above storage condition. Besides, please return the unused wells to the foil pouch containing the desiccant pack, and reseal along entire edge of zip-seal.|3. Expiration date is 6 months from shipment date.|4. It is highly recommended to use the remaining reagents within 1 month provided this is within the expiration date of the kit.