ROS has a very short half-lives in biological environment as they are influenced by exposure to ambient oxygen. As it is highly reactive and hard to measure care should be taken to ensure the stability of the sample during isolation, preparation, storage, and analysis.
ELISA is the most commonly used method of detecting and quantifying the concentration of an antigen in an unknown sample. During the experiment, If you get a weak signal, then make sure reagents are at room temperature before starting the assay. Try increasing incubation times to ensure maximal antibody binding and amplify the signal. Secondly, if you get values above 0 in the negative control indicates a high background signal. Try to consider reducing your antibody concentration and prevent non-specific binding of antibodies by using affinity-purified antibody and suitable blocking buffers. To avoid high well to well variation, do not stack plates during incubation, no bubbles in the plate and wash wells thoroughly to avoid variation.
Get tips on using Ambion™ RecoverAll™ Total Nucleic Acid Isolation Kit for FFPE to perform RNA isolation / purification Tissue - Rat Kidney
Get tips on using miRNeasy Mini kit to perform RNA isolation / purification Cells - primary hematopoietic stem cells
Get tips on using miRNeasy Mini kit to perform RNA isolation / purification Cells - primary human endometrial stromal cells
Get tips on using miRNeasy Mini kit to perform RNA isolation / purification Cells - primary human aortic smooth muscle cells
Fill out your contact details and receive price quotes in your Inbox
Outsource experiment