[3dem] Microtubule in vitro polymerisation

[Gregory Alushin]

Hi Linda-

We would also see those sorts of blobs from time to time in our negative stain images of microtubules.  Is batch-to-batch variation in tubulin the only thing that is changing between experiments?  We would sometimes include small amounts of NP-40 detergent in our buffers, the stock of which would oxidize with time after being opened and left in the fridge.  Sometimes swapping for a new bottle would solve this type of issue.  Not sure if this is helpful in your case, but non-tubulin buffer variation may be another thing to consider.

Good luck!
-Greg Alushin


From: 3dem <3dem-bounces at ncmir.ucsd.edu> on behalf of Linda Sandblad <linda.sandblad at umu.se>
Date: Thursday, November 9, 2017 at 4:33 PM
To: "3dem at ncmir.ucsd.edu" <3dem at ncmir.ucsd.edu>
Subject: [3dem] Microtubule in vitro polymerisation

Hi!! I must mention, at the EM facility we have many beautiful samples, it’s just that I often think the artifacts are more interesting to discuss ;-)
This is a question especially to all of you microtubule experts; When our students prepare mt from Cytoskeleton bovine tubulin it sometimes looks perfect, but sometimes it is all covered by those huge blobs, what do you think cause this artefact??
Happy to hear your thoughts! Best Linda
[cid:image001.png at 01D35985.CA0A4730]

Linda Sandblad
Department of Molecular Biology
Umeå University
Tel: +46 709324936

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