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<p class="MsoNormal"><span style="font-size:11.0pt;font-family:Calibri">Hi Linda-<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:Calibri"><o:p> </o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:Calibri">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.<o:p></o:p></span></p>
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<p class="MsoNormal"><span style="font-size:11.0pt;font-family:Calibri">Good luck!<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:Calibri">-Greg Alushin<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:Calibri"><o:p> </o:p></span></p>
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<p class="MsoNormal"><b><span style="font-family:Calibri;color:black">From: </span>
</b><span style="font-family:Calibri;color:black">3dem <3dem-bounces@ncmir.ucsd.edu> on behalf of Linda Sandblad <linda.sandblad@umu.se><br>
<b>Date: </b>Thursday, November 9, 2017 at 4:33 PM<br>
<b>To: </b>"3dem@ncmir.ucsd.edu" <3dem@ncmir.ucsd.edu><br>
<b>Subject: </b>[3dem] Microtubule in vitro polymerisation<o:p></o:p></span></p>
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<p class="MsoNormal" style="mso-margin-top-alt:auto;mso-margin-bottom-alt:auto">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 ;-)<o:p></o:p></p>
<p class="MsoNormal" style="mso-margin-top-alt:auto;mso-margin-bottom-alt:auto">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??<o:p></o:p></p>
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<span style="font-size:9.0pt;font-family:Helvetica;color:black">Happy to hear your thoughts! Best Linda<o:p></o:p></span></p>
<p class="MsoNormal" style="mso-margin-top-alt:auto;mso-margin-bottom-alt:auto"><img width="941" height="972" id="A570FC3E-DB41-4E42-89F7-DBBD2533D66B" src="cid:image001.png@01D35985.CA0A4730"><o:p></o:p></p>
<p class="MsoNormal" style="mso-margin-top-alt:auto;mso-margin-bottom-alt:auto"><o:p> </o:p></p>
<p class="MsoNormal" style="mso-margin-top-alt:auto;mso-margin-bottom-alt:auto">Linda Sandblad<o:p></o:p></p>
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<p class="MsoNormal"><span style="color:black">Department of Molecular Biology<br>
Umeå University</span><span style="font-family:PMingLiU;color:black"><br>
</span><span style="color:black">Tel: +46 709324936<o:p></o:p></span></p>
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