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Preprint

Western blot

NR Nasser M Rizk
MG Masoud Ghamari-Langroudi
IC Işın Çakır

Last updated date: Jun 28, 2022 Views: 626 Forks: 0

original An abbreviated version of this protocol was published in eLife
Sulforaphane reduces obesity by reversing leptin resistance
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Hi Wei,

 

Thanks for reaching out. The tissues were processed in the RIPA lysis buffer as published in the article. We use a TissuLyser from Qiagen for tissue homogenization. Antibody concentrations we typically use are:

 

p-STAT3: 1/1000

STAT3: 1/1000

Ac-aTubulin: 1/10,000

GAPDH: 1/10,000

 

We usually do the primary antibody incubations overnight at cold room and the secondaries at room temp for 1hr. Please let me know if you'd need more info on any of these.

 

Best wishes. -Isin

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How to cite：
Readers should cite both the Bio-protocol preprint and the original research article where this protocol was used:
  1. Rizk, N M, Ghamari-Langroudi, M and Çakır, I(2022). Western blot. Bio-protocol Preprint. bio-protocol.org/prep1750.
  2. Hadley, C. K., Pan, P. L., Rizk, N. M., Çakır, I., El-Gamal, A., Elsayegh, D., Fadel, A., Ghamari-Langroudi, M. and Mohamed, O. Sulforaphane reduces obesity by reversing leptin resistance. eLife. DOI: 10.7554/eLife.67368

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