Hi, I measured homocysteine levels in rat liver tissue homogenate with ELISA kit but I coldn't get results I think because of the homogenate's dansity, so I should dilute it. Is there anybody who have experience about measuring homocysteine levels in rat liver tissue with ELISA?
By the way, I am using CSB- E13376R CUSABİO Rat Homocysteine Elisa Kit (96 test). THANK YOU.
According to the manufacturer, you should use the following protocol:
Tissue Homogenates:
100 mg tissue was rinsed with 1XPBS,
homogenized in 1 ml of 1X PBS and stored overnight at
- 20°C. After two freeze - thaw cycles were performed to break the cell membranes, the homogenates were centrifuged for 5 minutes at 5000 x g, 2 -8°C. The supernate was removed and assayed immediately. Alternatively, aliquot
and store samples at - 20°C or - 80 °C. Centrifuge the sample again after thawing
before the assay. Avoid repeated freeze-thaw cycles.
However, it is mentioned on the web page that improved protocols might be available. You should definitely contact the manufacturer.
http://www.cusabio.com/uploadfile/Ins/2017-02-04/CSB-E13376r.pdf
Firstly, thank you for kind answer. According to this protocol, there is no need to extra dilution tissue homogenate; but there is something missing. I have talked them; they accept that tissue need extra dilution which is not writing in protocol. That is what I wanted to ask.
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