Protein extraction and Western blot analysis were performed as previously described (35 (link)). The antibodies: PTPN14 (1:1000 dilution), pYAP (s127, 1:1000 dilution), total YAP (1:1000 dilution), c-parp/parp (1:1000) and GAPDH (1:1000 dilution) were purchased from Cell Signaling Technology (Denvers, MA). Polyclonal goat anti-rabbit antibody (Cell Signaling Technology) and Western Blotting Detection System (Millipore) were used for exposure.
Polyclonal goat anti rabbit antibody
Manufactured by Cell Signaling Technology
Sourced in United States
Polyclonal goat anti-rabbit antibody is a secondary antibody produced in goats and designed to recognize and bind to rabbit primary antibodies. This product is commonly used in various immunoassay techniques to detect and quantify target proteins or analytes in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Western blotting detection system, by Merck Group (3 mentions)
Gadph antibody, by Abcam (2 mentions)
Pyaps127, by Cell Signaling Technology (1 mentions)
Ptpn14, by Cell Signaling Technology (1 mentions)
Total yap, by Cell Signaling Technology (1 mentions)
Gapdh, by Cell Signaling Technology (1 mentions)
Ab109520, by Abcam (1 mentions)
Western blotting detection system, by LI COR (1 mentions)
Ab8245, by Abcam (1 mentions)
Anti p65 antibody, by Abcam (1 mentions)
Anti sirt1 antibody, by Abcam (1 mentions)
Anti tnf α antibody, by Abcam (1 mentions)
Anti il 6 antibody, by Cell Signaling Technology (1 mentions)
Inos antibody, by Santa Cruz Biotechnology (1 mentions)
P nf κb antibody, by Cell Signaling Technology (1 mentions)
Ecl chemiluminescence reagent, by Cytiva (1 mentions)
5 protocols using polyclonal goat anti rabbit antibody
1
Protein Analysis via Western Blot
2
Protein Expression Analysis by Western Blot
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Protein extraction and Western blot analysis were performed referring to the literature step by step. Antibody: SIRT1, Runx2, OPN, OCN (1:1000, diluted, ab109520, abcam, USA), GAPDH (1:1000, diluted, ab8245, abcam, USA). The polyclonal goat anti-rabbit antibody (Cell Signaling Technology, MA, USA) and Western blotting detection system (Odyssey, LI-COR, LN, USA) were used for detection[22 (link)].
3
Western Blot Analysis of Inflammatory Markers
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The proteins in cells of each group were extracted and centrifuged at 4 degrees and then added to the protein lysate for cell lysis with RIPA lysis buffer-combined protease inhibitor cocktail at a ratio of 99 : 1. After that, the total protein was quantified with the BCA method.
The proteins were transferred to the PVDF membrane by electrophoresis with 12% SDS polyacrylamide gel at 300mA2h and then sealed at room temperature with 5% skim milk sealant and TBST for 1 h. The primary antibodies were anti-SIRT1 antibody (diluted 1 : 2000, Abcam, catalog no. AB32441), anti-P65 antibody (diluted 1 : 1500, Abcam, catalog no. AB16502), anti-TNF-α antibody (diluted 1 : 500, Abcam, catalog no. AB6671), anti-IL-6 antibody (diluted 1 : 2000, Cell Signaling Technology, catalog no. 5216), iNOS antibody (diluted 1 : 2000, Santa Cruz Biotechnology, catalog no. SC-650), and GADPH antibody (1 : 5000, Abcam, catalog no. ab9485). All these antibodies were diluted with closed solution, incubated at room temperature for 1 h, and washed with PBS buffer 3 times for 5 min. The polyclonal goat anti-rabbit antibody 1 : 5000 (Cell Signaling Technology) diluted in the closed solution was used as the secondary antibody. Then, it was detected with ECL chemiluminescence reagent imaging (Amersham) and western blotting detection system (Millipore).
The proteins were transferred to the PVDF membrane by electrophoresis with 12% SDS polyacrylamide gel at 300mA2h and then sealed at room temperature with 5% skim milk sealant and TBST for 1 h. The primary antibodies were anti-SIRT1 antibody (diluted 1 : 2000, Abcam, catalog no. AB32441), anti-P65 antibody (diluted 1 : 1500, Abcam, catalog no. AB16502), anti-TNF-α antibody (diluted 1 : 500, Abcam, catalog no. AB6671), anti-IL-6 antibody (diluted 1 : 2000, Cell Signaling Technology, catalog no. 5216), iNOS antibody (diluted 1 : 2000, Santa Cruz Biotechnology, catalog no. SC-650), and GADPH antibody (1 : 5000, Abcam, catalog no. ab9485). All these antibodies were diluted with closed solution, incubated at room temperature for 1 h, and washed with PBS buffer 3 times for 5 min. The polyclonal goat anti-rabbit antibody 1 : 5000 (Cell Signaling Technology) diluted in the closed solution was used as the secondary antibody. Then, it was detected with ECL chemiluminescence reagent imaging (Amersham) and western blotting detection system (Millipore).
4
Western Blot Analysis of Protein Signaling
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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).
5
Protein Extraction and Western Blot
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Protein extraction and Western blot analysis were performed as described previously (26) . The antibodies used in this study are listed in Supplementary Table S2. Polyclonal goat anti-rabbit antibody (Cell Signaling Technology) and Western Blotting Detection System (Millipore) were used for exposure.
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