上海武昊经贸有限公司
2012/12/19 17:42:36ELISA Kit for Advanced Glycation End Product （AGE） General shuo ming shu
[ INTENDED USE ]
The kit is a competitive inhibition enzyme immunoassay technique for the in vitro quantitative measurement of AGE in serum, plasma and other biological fluids.
[ STORAGE OF THE KITS ]
1. For unopened kit: All the reagents should be kept according to the labels on vials. The Standard, Detection Reagent A, Detection Reagent B and the 96-well strip plate should be stored at -20oC upon receipt while the others should be at 4 oC.
2. For opened kit: When the kit is opened, the remaining reagents still need to be stored according to the above storage condition. Besides, please return the unused wells to the foil pouch containing the desiccant pack, and reseal along entire edge of zip-seal.
Note:
It is highly recommended to use the remaining reagents within 1 month provided this is within the expiration date of the kit.
[ SAMPLE COLLECTION AND STORAGE ]
Serum - Allow samples to clot for two hours at room temperature or overnight at 4oC before centrifugation for 20 minutes at approximay 1000×g. Assay immediay or store samples in aliquot at -20oC or -80oC. Avoid repeated freeze/thaw cycles.
Plasma - Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples for 15 minutes at 1000×g within 30 minutes of collection. Remove plasma and assay immediay or store samples in aliquot at -20oC or -80oC. Avoid repeated freeze/thaw cycles.
Other biological fluids - Centrifuge samples for 20 minutes at 1000×g. Remove particulates and assay immediay or store samples in aliquot at -20oC or -80oC. Avoid repeated freeze/thaw cycles.
Note:
1. Samples to be used within 5 days may be stored at 4oC, otherwise samples must be stored at -20oC （≤1 month） or -80oC （≤2 months） to avoid loss of bioactivity and contamination.
2. Sample hemolysis will influence the result, so hemolytic specimen can not be detected.
3. When performing the assay, bring samples to room temperature.
[ SAMPLE PREPARATION ]
1. Uscn, Inc. is only responsible for the kit itself, but not for the samples consumed during the assay. The user should calculate the possible amount of the samples used in the whole test. Please reserve sufficient samples in advance.
2. Please predict the concentration before assaying. If values for these are not within the range of the standard
curve, users must determine the optimal sample dilutions for their particular experiments.
3. If the samples are not indicated in the manual, a preliminary experiment to determine the validity of the kit is
necessary.
4. Tissue or cell extraction samples prepared by chemical lysis buffer may cause unexpected ELISA results due to the impacts from certain chemicals.
5. Due to the possibility of mismatching between antigen from other origin and antibody used in our kits （e.g., antibody targets conformational epitope rather than linear epitope）, some native or recombinant proteins from other manufacturers may not be recognized by our products.
6. Influenced by the factors including cell viability, cell number or sampling time, samples from cell culture supernatant may not be detected by the kit.
7. Fresh samples without long time storage is recommended for the test. Otherwise, protein degradation and denaturalization may occur in those samples and finally lead to wrong results.
[ TEST PRINCIPLE ]
This assay employs the competitive inhibition enzyme immunoassay technique. A monoclonal antibody specific for AGE has been pre-coated onto a microplate. A competitive inhibition reaction is launched between biotin labeled AGE and unlabeled AGE （Standards or samples） with the pre-coated antibody specific for AGE. After incubation the unbound conjugate is washed off. Next, avidin conjugated to Horseradish Peroxidase （HRP） is added to each microplate well and incubated. The amount of bound HRP conjugate is reverse proportional to the concentration of AGE in the sample. After addition of the substrate solution, the intensity of color developed is reverse proportional to the concentration of AGE in the sample.
