Human Complement Factor D/ Adipsin ELISA Kit Human Complement Factor D/ Adipsin ELISA Kit Catalog Number: EK1290 Size: 48 Test, 96 Test For the quantitative determination of human Complement Factor D (CFD) 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 Complement Factor D/ Adipsin 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 Sample Preparation …………………………………………………………...

Human Complement Factor D/ Adipsin 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 Assay Buffer (1×)/culture medium to Blank well. Serum/Plasma: Add 90 μl Assay Buffer (1×) and 10 μl prediluted sample to the sample well. Cell Culture Supernates: Add 100 μl cell culture supernates to the sample well. (The dilution refers to the Sample Preparation on Page 6). Add 50 μl of diluted Detect Antibody...

Human Complement Factor D/ Adipsin ELISA Kit DESCRIPTION CFD is the prohormone precursor to insulin made in the beta cells of the islets of Langerhans, specialized regions of the pancreas. In humans, proinsulin is encoded by the INS gene. It is post-translationally cleaved into two chains (peptide A and peptide B) that are covalently linked via two disulfide bonds and one molecule of C-peptide. There is conservation of much of the structure of proinsulin among mammalian species. CFD binds the Insulin Receptor A isoform (IR-A) with high affinity resulting in cell proliferation and migration. It...

Human Complement Factor D/ Adipsin ELISA Kit MATERIALS PROVIDED (96 Test) Unopened kit should be stored at 2 - 8℃.  Human CFD Microplate (1 plate): 96-well polystyrene microplate (12 strips of 8 wells) coated with an antibody against human CFD.  Human CFD Standard (2 vials): Recombinant human CFD in a buffered protein base with preservatives; lyophilized.  Human CFD Detect Antibody (1 vial, 70 μl): Biotin-conjugate anti- human CFD detect antibody; 100× liquid.  Streptavidin-HRP (1 vial, 140 μl): 100× liquid.  Assay Buffer (10×) (1 bottle, 10 ml): PBS with 0.5 % Tween-20 and 5 % BSA.  Substrate...

Human Complement Factor D/ Adipsin 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...

Human Complement Factor D/ Adipsin 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....

Human Complement Factor D/ Adipsin ELISA Kit SAMPLE PREPARATION Normal serum and plasma samples require a 2,000-fold dilution. A suggested 2,000-fold dilution is two step dilution: first, 10 μl sample + 390 μl Assay Buffer (1×); next, 10 μl Mix + 490 μl Assay Buffer (1×). 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...