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Measuring BDNF in muscle using sandwich ELISA

BDNF ELISA Muscle

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#1 TurtleBob

TurtleBob

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Posted 09 January 2014 - 05:14 PM

Hello, 

 

I would like to measure BDNF in muscle. I am currently using a commercial (pre-coated) sandwich ELISA kit, however I seem to be running into issues that I believe to be the result of muscle having low BDNF levels. 

 

1) Serially dilution of samples does not result in a corresponding change in OD/concentration for MUSCLE. However, spinal cord samples and BDNF standards look fine.

 

2) I am working with a limited dilution range (for this reason, my serial dilution consists of only 2-3 samples). This is because muscle contains low amounts of BDNF and so I can't dilute very much. On the other hand, I must dilute otherwise I will have interference for antibody binding (due to components of the homogenizing buffer- NaCl and Triton).

 

3) If there is nonspecific binding occurring, is it possible to purify BDNF from my samples? I have heard this protein can be unstable.

 

 

Do you have any ideas/suggestions of why I may not be seeing a difference in my serial dilutions? Are there other ways of measuring BDNF at such low concentrations?

 

Your help is very appreciated. Please let me know if I can provide any clarifying information!







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