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Dear Dr. Gupta,Thank for your protocol.I have 2 questions:(1)...
HP
Hai Phung Oct 5, 2016
Dear Dr. Gupta,
Thank for your protocol.
I have 2 questions:
(1) serial serum dilution: to make the standard curve, could I dilute FITC-Dextran in WT-mice serum to 10, 20, 40, 80 ng/ml. Could you suggest the lowest concentration that could be still detectable? Read more 
Thank for your protocol.
I have 2 questions:
(1) serial serum dilution: to make the standard curve, could I dilute FITC-Dextran in WT-mice serum to 10, 20, 40, 80 ng/ml. Could you suggest the lowest concentration that could be still detectable?
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Jalaj Gupta Answered Oct 10, 2016
Institute for Research in Biomedicine (IRB Barcelona), Spain
Dear Hai Phung,
Thank you for your interest in our protocol.
1) In my experience, I could detect minimum of 400ng/ml in the mouse serum. In theory you can dilute the FITC-Dextran in WT mice serum but you should include standards up to 500-600ng/ml (eg: 10, 50, 100, 200, 400, 600 ng/ml). We used 125ng/ml as lowest standard, which was detected easily.
2) We always used 100ul diluted serum and could detect the FITC-dextran quite nicely but I think you can still dilute the serum to detect FITC-dextran and then multiply by dilution factor in order to get final concentrations.
I hope this will help you,
best,
Jalaj
Thank you for your interest in our protocol.
1) In my experience, I could detect minimum of 400ng/ml in the mouse serum. In theory you can dilute the FITC-Dextran in WT mice serum but you should include standards up to 500-600ng/ml (eg: 10, 50, 100, 200, 400, 600 ng/ml). We used 125ng/ml as lowest standard, which was detected easily.
2) We always used 100ul diluted serum and could detect the FITC-dextran quite nicely but I think you can still dilute the serum to detect FITC-dextran and then multiply by dilution factor in order to get final concentrations.
I hope this will help you,
best,
Jalaj
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