1 M sodium borate buffer solution

Txema

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.

JustDave

I used the protocol found here: neuro.bcm.edu/_web/jankowskylab/protocols/SB%20buffer.doc

karlkoeh

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!

JustDave

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.

Txema

Thanks! We have prepared it now and will make incubations soon. Hopefully we can post nice results. Thank you again for the support.

jroc093

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?

Shashank20

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