Human C-peptide ELISA Kit Human C-peptide ELISA Kit Catalog Number: EK1251 Size: 48 Test, 96 Test For the quantitative determination of human C-peptide concentrations in cell culture supernates, serum and plasma. This package insert must be read entirely before using this product. For proper performance, follow the protocol provided with each individual kit. MULTISCIENCES (LIANKE) BIOTECH, CO., LTD. 3F, Building B, 36 Xiang Mao Road, Gongshu District, Hangzhou, Zhejiang Province, China. www.multisciences.net Tel: +86 057128828618-88662 Fax: +86-0571-28828618 E-mail: [email protected]

Human C-peptide ELISA Kit TABLE OF CONTENTS ASSAY PROCEDURE SUMMARY ……………………………………… 1 Introduction Description ………………………………………………………………… 2 Principle of the Assay ……………………………………………………… 2 Limitations of the Procedure ……………………………………………… 2 General Information Materials Provided ………………………………………………………… 3 Storage … … … … … … … … … … … … … … … … … … … … … … … … … … 3 Other Supplies Required …………………………………………………… 4 Precaution …………………………………………………………………4 Technical Hints …………………………………………………………… 5 Assay Protocol Sample Collection and Storage …………………………………………… 5 Reagent Preparation ……………………

Human C-peptide ELISA Kit ASSAY PROCEDURE SUMMARY 1. Prepare all reagents and standards as directed. 2. Add 300 μl Washing Buffer (1×) per well to soak for about 30 seconds. Use immediately after aspirate. Add 100 μl 2-fold diluted Standard to Standard well. Add 100 μl Standard Diluent/culture medium to Blank well. Serum/Plasma: Add 90 μl Assay Buffer (1×) and 10 μl sample to the sample well. Cell Culture Supernates: Add 100 μl cell culture supernates to the sample well. Add 50 μl of diluted Detect Antibody to each well. Step 3, 4 and 5 should be completed within 15 minutes. Cover with an adhesive...

Human C-peptide ELISA Kit DESCRIPTION C-Peptide, or connective peptide, is secreted by Pancreatic beta cell. It shares a common proinsulin with the mature Insulin. It is a short 31-amino-acid polypeptide that connects insulin's A-chain to its B-chain in the proinsulin molecule. C-peptide has been shown to bind to the surface of a number of cell types such as neuronal, endothelial, fibroblast and renal tubular, at nanomolar concentrations to a receptor that is likely G-protein-coupled. C-peptide exerts vascular anti-inflammatory effects via the downregulation of endothelial cell surface adhesion...

Human C-peptide ELISA Kit MATERIALS PROVIDED (96 Test) Unopened kit should be stored at 2 - 8℃.  Human C-peptide Microplate (1 plate): 96-well polystyrene microplate (12 strips of 8 wells) coated with an antibody against human C-peptide.  Human C-peptide Standard (2 vials): Recombinant human C-peptide in a buffered protein base with preservatives; lyophilized.  Human C-peptide Detect Antibody (1 vial, 70 μl): HRP-conjugate anti- human C-peptide detect antibody; 100× liquid.  Standard Diluent (1 bottle, 5 ml): In some, very rare cases, an insoluble precipitate of stabilizing protein has been...

Human C-peptide ELISA Kit OTHER SUPPLIES REQUIRED  Microplate reader capable of measuring absorbance at 450 nm, with correction wavelength set at 570 nm or 630 nm.  Pipettes and pipette tips.  50 μl to 300 μl adjustable multichannel micropipette with disposable tips.  Multichannel micropipette reservoir.  Beakers, flasks, cylinders necessary for preparation of reagents.  Deionized or distilled water.  Polypropylene test tubes for dilution. PRECAUTION  All chemicals should be considered as potentially hazardous.  We therefore recommend that this product is handled only by those persons...

Human C-peptide ELISA Kit TECHNICAL HINTS  When mixing or reconstituting protein solutions, always avoid foaming.  To avoid cross-contamination, change pipette tips between additions of each standard level, between sample additions, and between reagent additions. Also, use separate reservoirs for each reagent.  When using an automated plate washer, adding a 30 seconds soak period before washing step and/or rotating the plate between wash steps may improve assay precision.  To ensure accurate results, proper adhesion of plate sealers during incubation steps is necessary.  Substrate Solution...

Human C-peptide ELISA Kit REAGENT PREPARATION Bring all reagents and samples to room temperature before use. If crystals form in the Buffer Concentrates, warm and gently stir them until completely dissolved. Washing Buffer (1×) Pour entire contents (50 ml) of the Washing Buffer (20×) into a clean 1,000 ml graduated cylinder. Bring to final volume of 1,000 ml with pure or deionized water. Mix gently to avoid foaming. Transfer to a clean wash bottle and store at 2 to 25℃. Washing Buffer (1×) is stable for 30 days. Assay Buffer (1×) Pour the entire contents (5 ml) of the Assay Buffer (10×) into...