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How to dilute in Quantikine ELISA (R&D systems)

Quantikine ELISA

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#1 SagoJ

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Posted 15 November 2013 - 06:22 AM

Hello!
I am trying to do a simple ELISA from kits (all quantikine ELISA kits from R&D system). I am doing the TNF (cat. noDTA00C) , Il-12 (p40) (cat. no DP400) and Il-10 (cat. no D1000B) ELISA. I have both serum and cell supernatant samples (from monocyte derived macrophages).

My problem is how to dilute the samples, in some kit there is a guideline, but not in some. For example, in the TNF kit (cat. DTA00C) they provide the reagent RD6-35, but no information on if I can dilute both supernatant and serum with it or how samples should be diluted. In the Il-12 kit, the reagents are RD5P for supernatant and RD6-13 for serum, but no information on dilution. In Il-10 this is RD5C and RD6P for supernatant and serum, respectively.
I wonder if you have any experience in this and how did you dilute your samples to stay within the standard curve?
Many many thanks for any answer! 



#2 tkf

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Posted 20 November 2013 - 11:50 AM

Where the kit does not provide relevant info, I would dilute the supernatant samples in the fresh culture medium that was used to grow the cells. For the serum samples, I would dilute in the same buffer used to prepare the standard curve.






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