Reg1b ELISA Kit (Human) (OKRC00848) Instructions for Use
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Background ................................................................................................................................. 2 2. Assay Summary .......................................................................................................................... 3 3. Storage and Stability ................................................................................................................... 3 4. Kit Components ........................................................................................................................... 3 5. Precautions.................................................................................................................................. 4 6. Required Materials Not Supplied ................................................................................................ 4 7. Technical Application Tips .......................................................................................................... 4 2. Reagent Preparation ................................................................................................................... 5 3. Assay Procedure ......................................................................................................................... 6 4. Calculation of Results ................................................................................................................. 7 5. Precision ...................................................................................................................................... 8 6. Technical Resources................................................................................................................... 8 www.avivasysbio.com–888.880.0001 – [email protected] 1 Reg1B ELISA Kit (Human) (OKRC00848) , Rev: March 3, 2020 1. Background Principle The Aviva Systems Biology PSPS2 ELISA kit (Human) is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of Human PSPS2 in serum, plasma, and cell culture supernatants. This assay employs an antibody specific for Human PSPS2 coated on a 96-well plate. Standards and samples are pipetted into the wells and PSPS2 present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-human PSPS2 antibody is added. After washing away unbound biotinylated antibody, HRP-conjugated streptavidin is pipetted to the wells. The wells are again washed, a TMB subst rate solution is added to the wells and color develops in proportion to the amount of PSPS2 bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm. Target Background REG1B might act as an inhibitor of spontaneous calcium carbonate precipitation. REG1B may be associated with neuronal sprouting in brain, and with brain and pancreas regeneration.This gene is a type I subclass member of the Reg gene family. The Reg gene family is a multigene family grouped into four subclasses, types I, II, III and IV based on the primary structures of the encoded proteins. This gene encodes a protein secreted by the exocrine pancreas that is highly similar to the REG1A protein. The related REG1A protein is associated with islet cell regeneration and diabetogenesis, and may be involved in pancreatic lithogenesis. Reg family members REG1A, REGL, PAP and this gene are tandemly clustered on chromosome 2p12 and may have arisen from the same ancestral gene by gene duplication. Publication Note: This RefSeq record includes a subset of the publications that are available for this gene. Please see the Entrez Gene record to access additional publications. General Specifications Human PSPS2 UniProt: P48304 Specificity GeneID: 5968 Target Alias: REG1B, REGL, Lithostathine-1-beta (Pancreatic stone protein 2) (PSP-2) (Regenerating islet-derived protein 1-beta) (REG-1-beta) (Regenerating protein I beta) Cross-Reactivity No detectable cross-reactivity with other relevant proteins www.avivasysbio.com–888.880.0001 – [email protected] 2 Reg1B ELISA Kit (Human) (OKRC00848) , Rev: March 3, 2020 2. Assay Summary 3. Storage and Stability • Open kit immediately upon receipt. Store components at -20°C (NOTE: exceptions below) for long term storage of 6 months until expiration date. Avoid any freeze/thaw cycles. 4. Kit Components • The following reagents are the provided contents of the kit. Description Quantity Storage Conditions PSPS2 Microplate 96 Wells (12 x 8 Well strips) PSPS2 Lyophilized Standard 2 vials Biotinylated PSPS2 Detector Antibody 2 vials -20°C for 6 months 500X Streptavidin-HRP Conjugate 1 x 200 uL 5X Sample Diluent 1 x 15 mL 20X Wash Buffer 1 x 25 mL Stop Solution 1 x 8 mL Store at 4°C for 6 Assay Diluent B 1 x 30 ml months TMB Substrate 1 x 12 mL www.avivasysbio.com–888.880.0001 – [email protected] 3 Reg1B ELISA Kit (Human) (OKRC00848) , Rev: March 3, 2020 5. Precautions • Read instructions fully prior to beginning use of the assay kit. • Any deviations or modifications from the described method or use of other reagents could result in a reduction of performance. • Reduce exposure to potentially harmful substances by wearing personal protective lab equipment including lab coats, gloves and glasses. • For information on hazardous substances included in the kit please refer to the Material Safety Data Sheet (MSDS). • Kit cannot be used beyond the expiration date on the label. 6. Required Materials Not Supplied • Microplate reader capable of reading absorbance at 450 nm. • Automated plate washer (optional). • Pipettes capable of precisely dispensing 0.5 µL through 1 mL volumes of aqueous solutions. • Pipettes or volumetric glassware capable of precisely measuring 1 mL through 100 mL of aqueous solutions. • New, clean tubes and/or micro-centrifuge tubes for the preparation of standards or samples. • Absorbent paper or paper toweling. • Distilled or deionized ultrapure water. 7. Technical Application Tips • Do not mix or substitute components from other kits. • To ensure the validity of experimental operation, it is recommended that pilot experiments using standards and a small selection of sample dilutions to ensure optimal dilution range for quantitation. • Samples exhibiting OD measurements higher than the highest standard should be diluted further in the appropriate sample dilution buffers. • Prior to using the kit, briefly spin component tubes to collect all reagents at the bottom. • Replicate wells are recommended for standards and samples. • Cover microplate while incubating to prevent evaporation. • Do not allow the microplate wells dry at any point during the assay procedure. • Do not reuse tips or tube to prevent cross contamination. • Avoid causing bubbles or foaming when pipetting, mixing or reconstituting. • Completely remove of all liquids when washing to prevent cross contamination. • Prepare reagents immediately prior to use and do not store, with the exception of the top standard. • Equilibrate all materials to ambient room temperature prior to use (standards exception). • Pipetting less than 1 µL is not recommended for optimal assay accuracy. • Once the procedure has been started, all steps should be completed without interruption. Ensure that all reagents, materials and devices are ready at the appropriate time. • Incubation times will affect results. All wells should be handled in the same sequential order and time intervals for optimal results. • Samples containing bilirubin, precipitates or fibrin strands or are hemolytic or lipemic might cause inaccurate results due to interfering factors. • TMB Substrate is easily contaminated and should be colorless or light blue until added to plate. Handle carefully and protect from light. www.avivasysbio.com–888.880.0001 – [email protected] 4 Reg1B ELISA Kit (Human) (OKRC00848) , Rev: March 3, 2020 Sample Preparation Guidelines 1.1 Sample Preparation and Storage • Samples must be tested to determine if the kit is valid. • Store samples to be assayed at 4°C for 24 hours prior being assayed. • For long term storage, aliquot and freeze samples at -20°C. Avoid repeated freeze-thaw cycles. General Sample Preparation Guidelines: • Serum - Use a serum separator tube and allow samples to clot for 30 minutes before centrifugation for 10 minutes at 1000x g, and detect; or aliquot and store samples at -20°C to -70°C (Stored at 2- 8°C if tested within 24 hours). Avoid freeze/thaw cycles. • Plasma - Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge for 15 minutes at 1000x g within 30 minutes of collection,and detect; or aliquot and store samples at -20°C to -70°C (Stored