The protein was separated in SDS-PAGE system according to the instructions of the existing literature (14 (link)). The protein antibodies used were as follows: P-NF-κB antibody (diluted at 1:1000; Cell Signaling Technology, catalog number: 9936T), p-MAPK (ERK1/2) antibody (diluted at 1:1000, Cell Signaling Technology, catalog number: 5235LF), p-TLR4 antibody (diluted at 1:500; Abcam, catalog number: ab13556) and GADPH antibody (diluted at 1:1000 dilution; Abcam, catalog number: ab9485). Polyclonal goat anti-rabbit antibody (Cell Signaling Technology) was used as a secondary antibody (at room temperature for 2 h), which was imaged with ECL chemiluminescence reagent (Amersham) and detected by Western blot (Millipore).
Protocol detail
Western Blot Analysis of Protein Signaling
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Anti antibody
Antibodies
Antibody
Chemiluminescence
Dilution
Erk1
Goat
Nf κb
Protein
Rabbit
Sds page
Western blot
Corresponding Organization :
Other organizations : Qiqihar Medical University, Mudanjiang Medical University
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Variable analysis
independent variables
- SDS-PAGE system
- P-NF-κB
- P-MAPK (ERK1/2)
- P-TLR4
- GADPH
- Protein antibodies used: P-NF-κB antibody (diluted at 1:1000; Cell Signaling Technology, catalog number: 9936T), p-MAPK (ERK1/2) antibody (diluted at 1:1000, Cell Signaling Technology, catalog number: 5235LF), p-TLR4 antibody (diluted at 1:500; Abcam, catalog number: ab13556) and GADPH antibody (diluted at 1:1000 dilution; Abcam, catalog number: ab9485)
- Polyclonal goat anti-rabbit antibody (Cell Signaling Technology) used as a secondary antibody (at room temperature for 2 h)
- ECL chemiluminescence reagent (Amersham) used for imaging
- Western blot (Millipore) used for detection
- No positive or negative controls were explicitly mentioned in the input.
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