Q: Which Oxidized LDL ELISA Kit should I use?
A: Our OxPL-LDL ELISA is specific for modifications found on the phospholipid component of LDL. Our other three OxLDL ELISA kits detect specific modifications on the protein and phospholipid components of LDL. Each uses a different capture antibody for specific modifications (CML, HNE, or MDA) and an anti-apoB antibody for detection. MDA is the most abundant modification found in LDL, but it is also the least stable. MDA-LDL is the ideal choice when testing fresh samples. Samples stored up to 6 months at -80ºC can be used to detect CML-LDL and/or HNE-LDL, which are more stable modifications.
Q: Do you have a recommendation for preparing cell lysates?
A: We have not tested our OxLDL ELISAs with cell lysates, but our recommendation is to prepare lysates in PBS alone or PBS with a low detergent concentration (such as <1% Triton). The standard should be prepared in the same buffer that is used to prepare the lysates. We don’t know how concentrated the LDL levels are in cell lysates or if enough LDL will be recovered after the precipitation step. A protein assay can be performed before and after the precipitation step to confirm that LDL is precipitated.
Q: Is oxLDL susceptible to degradation during sample storage?
A: All three modifications are susceptible to degradation over time. MDA is very unstable and can degrade after 1-2 months at -80ºC. CML and HNE modifications are more stable and samples can be stored at -80ºC for up to 6 months before significant degradation occurs.
Q: Can these kits be used with species other than human?
A: Unfortunately these products are not suitable for use with other species because an anti-human ApoB antibody is used for detection.