We need to evaluate the cytokines concentrations in brainstem [nucleus tractus solitarius (NTS)] of rats by Luminex Assay.
We have the analyte kit and the punches of the NTS.
We are not sure about sample preparation.
What is the best dilution (volume) of central nervous system tissues? What is the best diluent in this case?
I did some kind of luminex for CNS tissue. Rather than using the whole brain tissue, I purified lymphocytes first and then following the protocol for Luminex assay. Here is a link that may help you on this procedure.
Article Isolation of Brain-infiltrating Leukocytes
If you have no idea about where your analyte will fall on the standard curve provdied by the assay, the best thing to do is to analyze it using two dilutions: neat and a 1:10 dilution.
As for the diluent... when you say "punches" are these tissue? If you haven't lysate the tissue, the diluent is just simply whatever buffer you used in the lysate.
Hope that helps.
What is the best dilution (volume) of central nervous system tissues?
- You can try some dilutions with a few (positive) samples by performing a Bradford assay and using some cheaper cytokine assay. Tip: start trying 30, 50, and 100 ul of buffer for each sample.
What is the best diluent in this case?
- The manufacter may have written about sample preparation on the datasheet. If not, you can homogenize in the buffer provided in the kit. If it's not supplied, ask the manufacter about the recipe.
Hope that helps.
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