Biolegend
Mouse TNF-α ELISA MAX™ Standard
430902
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Used this kit to assess TNF-alpha levels in the serum and in cell culture supernatants. The assay was pretty easy to perform and the assay had a nice range of detection.
ELISA
Mouse serum samples and cell culture supernatants from mouse bone-marrow derived macrophages
Pre-working day (24h before working).Dilute your Capture antibody in the Coating buffer and add 100 microliter/well to the 96 well/plate.Next day, Wash plate and block it for 1 hour with Assay diluent. After the blocking time wash and add your samples and standards and incubate for 2h in RT (room temperature). After 2h of incubation wash again and add the diluted Detection antibody (incubate for 1h). Then wash again and add the enzyme (diluted in Assay diluent) and incubate for 30 min., then wash again and add the TMB Substrate solution. In the end use the Stop solution. Read the plate in a plate reader
Careful when pipetting to avoid cross contamination.
Was able to detect levels of cytokines reliably in serum and cell culture supernatants.
N/A
Was the simple and easy steps
Needs a big sample volume
Is highly easy to use, and very reliable. Also was cheap to perform when you are running a large number of samples.