Western Blot and Semiquantitative Analysis

The Western blot protocol and semiquantitative analysis were carried out following the protocol of our previous studies54 (link)56 (link). The following antibodieswere used: IL-1β antibody (rabbit monoclonal antibody, dilution 1:20000, Abcam.UK), TNF-α antibody (rabbit polyclonal antibody, dilution 1:100, Abcam. UK), VEGF antibody (rabbit monoclonal antibody, dilution 1:1000, Abcam. UK), HIF-1α antibody (rabbit monoclonal antibody, dilution 1:1000, Abcam. UK), MMP-1 antibody (rabbit monoclonal antibody, dilution 1:1000, Abcam. UK), MMP-13 antibody (rabbit monoclonal antibody, dilution 1:500, Abcam. UK), CHRM1 antibody (rabbit polyclonal antibody, dilution 1:500, Abcam. UK), CHRM3 antibody (rabbit monoclonal antibody, dilution 1:1000, Abcam. UK), CHRNA2 antibody (rabbit monoclonal antibody, dilution 1:1000, Abcam. UK), NR3C1 antibody (rabbit monoclonal antibody, dilution 1:50000, Abcam. UK). All experiments were done in triplicate. Mean normalized protein expression ± SEM was calculated from independent experiments.

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Zhang Y., Bai M., Zhang B., Liu C., Guo Q., Sun Y., Wang D., Wang C., Jiang Y., Lin N, & Li S. (2015). Uncovering pharmacological mechanisms of Wu-tou decoction acting on rheumatoid arthritis through systems approaches: drug-target prediction, network analysis and experimental validation. Scientific Reports, 5, 9463.

Publication 2015

IL-1β antibody TNF-α antibody VEGF antibody HIF-1α antibody MMP-13 antibody

Antibody Chrm1 Chrm3 Dilution Il 1β Mmp 1 Mmp 13 Monoclonal antibody Protein Rabbit Tnf α Vegf Western blot

Corresponding Organization :

Other organizations : Chinese Academy of Medical Sciences & Peking Union Medical College, Tsinghua University, Tianjin International Joint Academy of Biomedicine

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Protocol cited in 6 other protocols

Variable analysis

independent variables

None explicitly mentioned

dependent variables

IL-1β protein expression
TNF-α protein expression
VEGF protein expression
HIF-1α protein expression
MMP-1 protein expression
MMP-13 protein expression
CHRM1 protein expression
CHRM3 protein expression
CHRNA2 protein expression
NR3C1 protein expression

control variables

None explicitly mentioned

controls

Positive controls: None mentioned
Negative controls: None mentioned

Annotations

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