Human IDO1 ELISA Kit
Cat.No. RJ-15030
For the quantitative in vitro determination of Human indoleamine 2,3-dioxygenase 1 concentrations in
serum - plasma - tissue homogenates - other biological fluids
FOR LABORATORY RESEARCH USE ONLY.
NOT FOR USE IN DIAGNOSTIC PROCEDURES.
This package insert must be read in its entirety before using this product.
ELISA
ENZYME LINKED IMMUNOSORBENT ASSAY
INTENDED USE AND TEST PRINCIPLE
This IDO1 ELISA kit is intended Laboratory for Research use only and is not for use in diagnostic or therapeutic procedures. The Stop Solution changes the color from blue to yellow and the intensity of the color is measured at 450 nm using a spectrophotometer. In order to measure the concentration of IDO1 in the sample, this IDO1 ELISA Kit includes a set of calibration standards. The calibration standards are assayed at the same time as the samples and allow the operator to produce a standard curve of Optical Density versus IDO1 concentration. The concentration of IDO1 in the samples is then determined by comparing the O.D. of the samples to the standard curve.
SAMPLE COLLECTION AND STORAGES
Serum - Use a serum separator tube and allow samples to clot for two hours at room temperature or overnight at 4℃ before centrifugation for 20 minutes at approximay 1000×g. Assay freshly prepared serum immediay or store samples in aliquot at -20℃ or -80℃ for later use. Avoid repeated freeze/thaw cycles.
Plasma - Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples for 15 minutes at 1000×g at 2-8℃ within 30 minutes of collection. Remove plasma and assay immediay or store samples in aliquot at -20℃ or -80℃ for later use. Avoid repeated freeze/thaw cycles.
Tissue homogenates - For general information, hemolysis blood may affect the result, so you should rinse the tissues with ice-cold PBS (0.01M, pH=7.4) to remove excess blood thoroughly. Tissue pieces should be weighed and then minced to small pieces which will be homogenized in PBS (the volume depends on the weight of the tissue. 9mL PBS would be appropriate to 1 gram tissue pieces. Some protease inhibitor is recommended to add into the PBS.) with a glass homogenizer on ice. To further break the cells, you can sonicate the suspension with an ultrasonic cell disrupter or subject it to freeze-thaw cycles. The homogenates are then centrifugated for 5minutes at 5000×g to get the supernate.
Cell culture supernates and other biological fluids - Centrifuge samples for 20 minutes at 1000×g. Remove particulates and assay immediay or store samples in aliquot at -20℃ or -80℃ for later use. Avoid repeated freeze/thaw cycles.
Note: The samples shoule be centrifugated dequay and no hemolysis or granule was allowed.
MATERIALS REQUIRED BUT NOT SUPPLIED
1. 37 ℃ incubator
2. Standard microplate reader capable of measuring absorbance at 450 nm
3. Precision pipettes, disposable pipette tips and Absorbent paper
4. Distilled or deionized water
REAGENTS PROVIDED
All reagents provided are stored at 2-8°C. Refer to the expiration date on the label.
Name | 96 determinations | 48 determinations |
MICROTITER PLATE | 8*12strips | 8*6strips |
STANDARD(6 vial) | 0.3ml/vial | 0.3ml/vial |
SAMPLE DILUENT | 6.0ml | 3.0ml |
ENZYME CONJUGATE | 10.0ml | 5.0ml |
WASH SOLUTION | 25ml | 15ml |
SUBSTRATE A | 6.0ml | 3.0ml |
SUBSTRATE B | 6.0ml | 3.0ml |
STOP SOLUTION | 6.0ml | 3.0ml |
Closure plate membrane | 2 | 2 |
User manual | 1 | 1 |
Sealed bags | 1 | 1 |
Note:
1. Standard concentration was followed by: 24, 12, 6, 3, 1.5, 0.75 IU/mL.
2. If samples generate values higher than the highest standard, please dilute the samples with Sample Diluent and repeat the assay.
PRECAUTIONS
REAGENT PREPARATION AND STORAGE
Wash Solution (1X) - Dilute 1 volume of Wash solution (20X) with 19 volumes of deionized or distilled water. Wash Solution is stable for 1 month at 2-8°C.