1 M sodium borate buffer solution

We have obtained our first partially cleared samples and now we wanted to try immunofluorescence on those samples. We have difficulties dissolving and preparing the 1 M sodium borate buffer solution, pH 8.5. Could somebody post a recipe for preparing it? Thank you in advance.

Comments

  • I used the protocol found here: neuro.bcm.edu/_web/jankowskylab/protocols/SB%20buffer.doc
  • Bear in mind that those directions are for a ~2 M solution. For 1 M, use 61.83 g boric acid and roughly 10 g NaOH to start with. Dissolve in 700-800 mL, pH to 8.5, and Q.S. to 1 L. With a little heat it should take about 15-20 minutes to dissolve the boric acid and NaOH. Good luck!
  • Yes, forgot to mention that. What they call 50x stock is really 2M. Didn't have any difficulty getting it to dissolve with a little heat and time as you noted.
  • TxemaTxema Posts: 2
    Thanks! We have prepared it now and will make incubations soon. Hopefully we can post nice results. Thank you again for the support.
  • Is the 1M boric acid solution made in 0.1% triton-x PBS solution? Thats what it looks like in the protocol.
    After staining and washing, do you go straight to focusclear? Does the brain need to be washed in focusclear for a few days and then fresh focusclear used for imaging?
  • Hi Everyone, for the staining of ETC cleared brains are you using 1M sodium Borate buffer or 0.5M sodium borate buffer. In the paper they mentioned 0.5M for the intact mouse brain tissue. Any suggestions ? Thanks
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