乙酰化和非乙酰化检测试剂盒的差异:

目前市场上的 cAMP 和 cGMP 检测试剂盒的一个关键步骤是将乙酸酐含与cAMP或 cGMP的样品一起孵育，以形成乙酰化的 cAMP 或 cGMP 衍生物。然后抗体与乙酰化 cAMP 或 cGMP 中间体结合,以提高灵敏度。众所周知，醋酸酐是一种毒性很强的强乙酰化剂, 长期接触会损害科研人员的健康。乙酸酐还可以乙酰化ATP、GTP或其它的芳环。乙酰化产物也可以被抗体识别，从而降低了灵敏性。
Product Highlight

• Based on the unique mouse monoclonal anti-cAMP antibody
• The only assay kit on the market with monoclonal anti-cAMP antibody (all other kits are based on polyclonal anti-cAMP antibodies)
• The monoclonal anti-cAMP antibody displays much higher selectivity over cGMP and ATP than polyclonal anti-cAMP antibody
• Avoids the batch-to-batch variations associated with polyclonal antibody productions from animals, thus providing the reproducibility

 
Principle Outline
NewEast Biosciences cAMP ELISA Kit is a competitive immunoassay to measure the cAMP levels, either from cell extracts or from in vitro adenylyl cyclase assays. Briefly, multi-well plates are coated with goat-anti-mouse serum. cAMP in cell extracts or in in vitro adenylyl cyclase assays will competitively bind to the monoclonal anti-cAMP antibody in the presence of fixed amounts of cAMP-conjugated horse-radish peroxidase or alkaline phosphatase. Known amounts of cAMP are used as standards to generate the calculation curve. After a short incubation, the excess reagents are washed away and substrate is added. The multiwell plates are then read on a microplate reader at 450 nm or 405 nm. The intensity of the yellow color is inversely proportional to the concentration of cAMP in samples. The measured optical density is used to calculate the concentration of cAMP in samples based on the curve from the cAMP standards.

NewEast Biosciences 花了两年时间筛选大量的单克隆抗体，从中筛出识别裸 cAMP 或 cGMP抗体，而不识别 ATP 和 GTP。因此，本公司试剂盒省去了使用有毒乙酸酐的乙酰化步骤, 提高的选择性并缩短了检测时间。