Q: What structures are detected with your AGE Competitive ELISA Kit?
A: Our AGE Competitive ELISA Kit uses an anti-AGE polyclonal antibody that detects multiple AGE structures. The AGE-BSA standard was prepared by reacting BSA with glycolaldehyde, followed by extensive dialysis and column purification. It contains CML, pentosidine and other AGE structures, but not CEL or methylglyoxal (MG). This same AGE-BSA was used as an antigen to prepare the anti-AGE polyclonal antibody provided in the kit.
Q: Are EDTA plasma samples compatible with your kits?
A: EDTA will not interfere with the Competitive AGE ELISA and any anticoagulant can be used when preparing plasma samples.
Q: Is your AGE Competitive ELISA Kit compatible with any species?
A: Our AGE ELISA Kit is not species specific and can be used with samples from any species.
Q: Is the AGE Competitive ELISA kit compatible with tissue lysates?
A: Tissue samples are compatible with this assay and lysates can be prepared in any lysis buffer.
Q: Can this ELISA be used to detect AGE in food samples?
A: Any protein sample can be used with our AGE ELISA Kit, including food samples, as long as proteins can be isolated from the sample.
Q: What cell number should I use in the ELISA?
A: Unfortunately we do not have a recommended cell number to use when preparing lysates, because this will depend on the AGE level of the sample, which will be different for each researcher. Our recommendation is to start with the most concentrated sample possible and prepare further dilutions later, if necessary, after running a small scale sample titration against the standard curve.
Q: What is the recommended protocol for preparing lysates?
A: To prepare lysates, we recommend resuspending cell or tissue samples in PBS containing proteinase inhibitors and optional 0.005% Butylated hydroxytoluene (a generic antioxidant available from Sigma-Aldrich; you can prepare 5% stock of BHT in methanol). Break up cells or tissues by homogenization or sonication, centrifuge at 12000 xg for 10 min and harvest the supernatant as your lysate.