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Western Blotting

DP Delayna Paulson
RH Rebecca Harms
CW Cody Ward
ML Mackenzie Latterell
GP Gregory J. Pazour
DF Darci M. Fink
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Immortalized mouse mesenteric lymphatic endothelial cells from confluent 9.6 cm2 wells (six-well plates) were lysed using RIPA buffer (Thermo Fisher Scientific, 89900) plus 1X Halt protease and phosphatase inhibitors with EDTA (Thermo Fisher Scientific) and cleared by centrifugation. Cell lysates were subjected to Western blot analysis using the XCell IITM Blot Module system (Invitrogen, EI9051). PVDF membranes were probed with primary antibodies against PROX-1 and β-actin as a loading control followed by Licor IRDye secondary antibodies. Membranes were imaged using a LI-COR Odyssey CLx imager with near-infrared Western blot detection and analyzed using ImageStudioLite software.

Copyright and License information:  ©2021 Paulson, Harms, Ward, Latterell, Pazour and Fink.
This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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