Goat anti –Monkey Interleukin 8
Collect Sample – serum or blood plasma
Package size: 96 determinations Catalogue No.: QRCT –657452EIA \ UTL
Storage: 2 - 8 °C
I. INTRODUCTION
The IL-8 is main to be produced by single cell. Other such as fiber cell, top skin cell, inside skin cell, liver the cell etc. is may also under the proper incitement term of to produce the IL-8.The IL-8 molecular weight is roughly 8 kD, primarily the live form is 72 amino acidses.The IL-8 amino acids sequence and many factor contain higher , belong to the same of household;Now first step proof the IL-8 households( as well call the PF4 household) at least contain 12 members.The IL-8 can be divided into the α with the β the gene for gene for, second cluster of α locating the second cluster of top, β of the No.4th chromosome of second cluster locate the 7th chromosome the top.Suffering of IL-8 also have two type:A kind of for can with IL-8 combinatively, but another kind can still join togethering other factor;Cell of neutral grain with love a cell surface to all expresses the abundant IL-8 to suffer. The IL-8 main biology activity is an attractions with arouse the cell of live and neutral grain, and once is named aftered the cell of neutral grain to arouse the live ( NAF), grain the cell the ( GCP), neutral grain the cell to arouse the live factor( NAF) etc.. The cell and IL-8 of neutral grain get in touch withes the empress occurrence the appearance the variety, and the definite direction rove around the reaction part and release a series of and live thing;The that these functions can cause the machine the part respond.
II. REAGENTS
Materials provided with the kits :
1、Coated Microtitration Strips 96 wells.
2、Standard 25, 50, 100, 200, 400 pg/ml, 1 set.
Enzyme Conjugate Solution, 12 ml.
A, 6 ml.
B, 6 ml.
Stopping Solution (1N HCl), 6 ml.
Rinsing buffer,60 ml( x 20).
dilution,15ml(x5)
Materials required but not provided :
Precision pipettes: 0.10, 0.20, and 1.0 ml.
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FOR INFORMATIONAL USE ONLY ? DO NOT USE FOR PERFORMING ASSAY ? REFER TO MOST CURRENT PACKAGE INSERT ACCOMPANYING TESTKIT
Goat anti –Monkey Interleukin 8
Collect Sample – serum or blood plasma
Package size: 96 determinations Catalogue No.: QRCT –657452EIA \ UTL
Storage: 2 - 8 °C
ortex mixer or equivalent.
Absorbent paper or paper towels.
Microtiter plate shaker
Graph paper.
A microtiter plate reader with a bandwidth of 10nm or less and an optical density range of 0-2 OD or greater at 450 nm .
III. STORAGE OF TEST KIT
Unopened test kits should be stored at 2-8°C upon receipt and the microtiter plate should be kept in a sealed bag with desiccants to minimize exposure to damp air. Opened test kits will remain stable until the expiration date shown, provided it is stored as described above. A microtiter plate reader with a bandwidth of 10nm or less and an optical density range of 0-2 OD or greater at 450nm wavelength is acceptable for use in absorbance measurement. All reagents should be allowed to reach room temperature before use.
IV. ASSAY PROCEDURE
STEP1 Prepare the ringing buffer by diluting the ringing buffer 20×(example: 60ml+1140ml H2O), Prepare the Sample Diluent by diluting the Diluent 5×(example: 15ml+60ml H2O)
STEP2 Dilute samples 1:100 distribing 5ul of sample into 500 ul of dilluent .
STEP3 Place 100ul of diluted sample /reagent blank/standard/controls to the wells of the strips. Incubate for 30 min at 37℃。Wash 5 times。
STEP4 Add 100ul of Enzyme Conjugate to each well。 Incubate for 30 min at 37℃。Wash 5 times。
STEP5 Add 50ul Substrate A and Substrate B to each well。Incubate for 15 min at 37℃。
STEP6 Add 50ul of stop solution。read absorbance at 450nm within 15 min
V. Important Notes
The wash procedure is critical. Insufficient washing will result in poor precision and falsely elevated absorbance readings. It is recommended that if manual pipetting is used, no more than 32 wells be used for each assay run, since pipetting of all standards, specimens and controls should be
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FOR INFORMATIONAL USE ONLY ? DO NOT USE FOR PERFORMING ASSAY ? REFER TO MOST CURRENT PACKAGE INSERT ACCOMPANYING TESTKIT
Goat anti –Monkey Interleukin 8
Collect Sample – serum or blood plasma
Package size: 96 determinations Catalogue No.: QRCT –657452EIA \ UTL
Storage: 2 - 8 °C
completed within 3 minutes. A full plate of 96 wells may be used if automated pipetting is available. Duplication of all standards and specimens, although not required, is recommended.
VI. CALCULATION OF RESULTS
Calculate the average absorbance values (A450) for each set of reference standards, control, and samples. Construct a standard curve by plotting the mean absorbance obtained for each reference standard against its concentration on linear graph paper, with absorbance on the vertical (y) axis and concentration on the horizontal (x) axis.Using the mean absorbance value for each sample, determine the corresponding concentration of in from the standard curve.
VII. EXAMPLE OF STANDARD CURVE
Results of a typical standard run with optical density readings at 450nm shown in the Y axis against concentrations shown in the X axis. This standard curve is for the purpose of illustration only, and should not be used to calculate unknowns. Each user should obtain his or her own data and standard curve.
VIII. CALCULATION OF RESULTS
The minimum detectable concentration of IL-8 in this assay is estimated to be 1.0 pg/ml.
IX. LIMITATIONS OF THE PROCEDURE
Reliable and reproducible results will be obtained when the assay procedure is carried out with a complete understanding of the package insert instructions and with adherence to good laboratory practice. The wash procedure is critical. Insufficient washing will result in poor precision and falsely elevated absorbance readings. The results obtained from the use of this kit should be used only as an adjunct to other diagnostic procedures and information available to the physician.
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