B1–B3, but not B4 beads bind to HMGB1.
Zhongliang Ju
Sangeeta S. Chavan
Daniel J. Antoine
Meghan Dancho
Teá Tsaava
Jianhua Li
Ben Lu
Yaakov A. Levine
Andrew Stiegler
Yehuda Tamari
Yousef Al-Abed
Jesse Roth
Kevin J. Tracey
Huan Yang
10.1371/journal.pone.0103992.g002
https://plos.figshare.com/articles/figure/_B1_8211_B3_but_not_B4_beads_bind_to_HMGB1_/1140859
<p>A) Depletion curve of HMGB1 binding to various DNA beads. Recombinant HMGB1 (2 µg per reaction) was mixed with a range of concentrations of different types of DNA beads as indicated and incubated at room temperature for two hours. The mixture was then centrifuged at 2,000 rpm for five minutes to precipitate the beads. Beads were then washed five times with PBS. Both supernatants and elute from beads were subjected to western blot analysis probed with anti-HMGB1 antibody. B) Saturation curve of HMGB1 binding to various DNA beads. Fixed amounts (20 µl drained beads) of B1–B3 or control beads were incubated with increasing amounts of HMGB1 (in 50 µl volume) at concentrations indicated for two hours at room temperature with rotation. The mixture was then centrifuged at 2,000 rpm for five minutes to precipitate the beads. The supernatants were aspirated, both supernatants and elute from beads were subjected to western blot for HMGB1 measurement using monoclonal anti-HMGB1 antibodies. The binding of 1 µg HMGB1 requires about 0.4 ng (B1 and beads) or 2.8 ng (B3) in beads, respectively. C) Time-course of HMGB1 binding to various DNA beads. B1–B3 beads (20 µl) were incubated with 500 ng of HMGB1 (50 µl total volume) at room temperature for the time periods indicated. HMGB1 bound to the beads was revealed by western blot analysis. Data are representative from 3–4 separate experiments.</p>
2014-08-15 03:35:15
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Sequestering HMGB 1
sequester HMGB 1
HMGB 1
Neutralizing HMGB 1 activity
coliti
DNA binding protein
High mobility group box 1
B 2 DNA beads
ibd
B 2 beads
il
bind HMGB 1