Immunoblotting

SR Seongmi K. Russell
JH Jessica K. Harrison
BO Benjamin S. Olson
HL Hyung Joo Lee
VO Valerie P. O’Brien
XX Xiaoyun Xing
JL Jonathan Livny
LY Lu Yu
ER Elisha D. O. Roberson
RB Rajdeep Bomjan
CF Changxu Fan
MS Marina Sha
SE Shady Estfanous
AA Amal O. Amer
MC Marco Colonna
TS Thaddeus S. Stappenbeck
TW Ting Wang
TH Thomas J. Hannan
SH Scott J. Hultgren
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Cells were lysed with cell lysis buffer (Cell Signaling, 9803S) according to the manufacturer’s instructions. Rapid Gold BCA Protein Assay kit was used to determine protein concentrations in the cell lysate, and equal amounts of protein were separated by SDS–PAGE and transferred to a nitrocellulose membrane. Membranes were incubated overnight with primary antibodies against caspase-1 (AdipoGen, AG-20B-0042-C100, 1:5,000) and β-actin (MA5-15739, Invitrogen, 1:10,000). HRP-linked secondary antibody (Cell Signaling, 7076S, 1:3,000) and ECL reagent (Amersham, RPN2209) were used to visualize protein bands.

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