Human CXCL1/KC ELISA Kit Catalog Number: EK196 Size: 48 Test, 96 Test For the quantitative determination of human chemokine (C-X-C motif) Ligand 1 (CXCL1/KC) 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]

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 ………………………………………………………… 6 Re

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 100 μl prediluted sample to the sample well. Cell Culture Supernates: Add 100 μl cell culture supernates to the sample well. Step 3 and 4 should be completed within 15 minutes. Incubate for 1.5 hours at RT. Aspirate and wash 6 times. Add 100 μl of diluted Detect Antibody to each well. Incubate...

DESCRIPTION The chemokine (C-X-C motif) ligand 1 (CXCL1) is a small cytokine belonging to the CXC chemokine family that was previously called GRO1 oncogene, GROα, KC and neutrophil-activating protein 3 (NAP-3). In humans, this protein is encoded by the CXCL1 gene. CXCL1 is secreted by human melanoma cells, has mitogenic properties and is implicated in melanoma pathogenesis. CXCL1 is expressed by macrophages, neutrophils and epithelial cells, and has neutrophil chemoattractant activity. This chemokine elicits its effects by signaling through the chemokine receptor CXCR2. CXCL1 plays a role in...

MATERIALS PROVIDED (96 Test) Unopened kit should be stored at 2 - 8℃.  Human CXCL1 Microplate (1 plate): 96-well polystyrene microplate (12 strips of 8 wells) coated with an antibody against human CXCL1.  Human CXCL1 Standard (2 vials): Recombinant human CXCL1 in a buffered protein base with preservatives; lyophilized.  Human CXCL1 Detect Antibody (1 vial, 70 μl): Biotin-conjugate anti- human CXCL1 detect antibody; 100× liquid.  Standard Diluent (1 bottle, 5 ml): In some, very rare cases, an insoluble precipitate of stabilizing protein has been seen in the Standard Diluent. This precipitate...

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 who have been trained in...

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 should remain colorless...

SAMPLE PREPARATION Normal serum and plasma samples require a 50-fold dilution. A suggested 50-fold dilution is 10 μl sample + 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 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℃....