For co-immunoprecipitations, cells were lysed with 1% NP40 NET buffer (Promoter Company, China), the supernatants were incubated overnight at 4°C with protein G-Sepharose beads (Sigma) conjugated with rabbit anti-Flag or anti-HA antibody, and then the samples were prepared for western blot assay. For western blot, the membrane was incubated overnight at 4°C with primary antibodies in Supplemental Table 2 , followed by anti-mouse or -rabbit IgG (1:3,000; Sigma, CA), and the signals were detected with an ECL assay kit (Amersham, Buckinghamshire, UK).
Anti mouse or rabbit igg
Manufactured by Merck Group
Sourced in United Kingdom
The Anti-mouse or -rabbit IgG product is a laboratory reagent used to detect and quantify the presence of mouse or rabbit immunoglobulin G (IgG) in biological samples. It functions as a secondary antibody, binding to the primary antibody that is specific to the target antigen. This interaction can be detected and measured, allowing researchers to analyze the levels of mouse or rabbit IgG in their experiments.
Automatically generated - may contain errors
Lab products found in correlation
Protein g sepharose beads, by Merck Group (2 mentions)
Ecl assay kit, by Cytiva (2 mentions)
Ab32360, by Abcam (1 mentions)
Anti α sma bm0002, by Boster Bio (1 mentions)
Anti phospho akt, by Cell Signaling Technology (1 mentions)
Anti phospho erk1 2, by Cell Signaling Technology (1 mentions)
Anti akt, by Santa Cruz Biotechnology (1 mentions)
Anti erk1 2, by Cell Signaling Technology (1 mentions)
3 protocols using anti mouse or rabbit igg
1
Co-immunoprecipitation and Western Blot Analysis
2
Investigating Signal Transduction Pathways
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Cell extracts and live tissues were digested in 1x RIPA buffer containing phosphatase inhibitor and PMSF (Goodbio Technology Co. Ltd. Wuhan, China). For western blot, the membrane was incubated overnight at 4 °C with primary antibodies: anti-p38 (ab195049, Abcam); anti-c-Jun (9165p, Cell Signaling Technology); anti-α-SMA (BM0002, Boster); anti-SATB1(3650, Cell Signaling Technology); anti-Sp1(10915, santa cruz); anti-Akt (21054, signalway antibody); anti-phospho-Akt (11054, signalway antibody); anti-ERK1/2 (1016, Cell Signaling Technology); anti-phospho-ERK1/2 (9101, Cell Signaling Technology), anti-phospho-p38 (11581, signalway antibody); anti-rabbit IgG-HRP (Promoter Biotechnology Ltd, China); anti-NFKB1 (ab32360, Abcam). Then followed by anti-mouse or -rabbit IgG (1:3,000; Sigma, CA), and the signals were detected with an ECL assay kit (Amersham, Buckinghamshire, UK). IL-6 (EHC007, NeoBioscience) and CTGF (900-K317, peprotech) enzyme-linked immunobsorbent assay (ELISA) were performed following the instructions by the manufacturer.
3
Western Blot and Co-immunoprecipitation Analysis
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Following standard electrophoresis, the membranes were probed overnight at 4 °C with primary antibodies listed in Supplementary Table 2. Subsequently, the membranes were incubated with anti-mouse or -rabbit IgG (1:3000; Abbkine, CA, USA), and the signals were detected using an ECL assay kit (Amersham, Buckinghamshire, UK).
For co-immunoprecipitations, the cells were lysed with 1% NP40 NET buffer (Promoter Company, China), the supernatants were incubated overnight at 4 °C with protein G-Sepharose beads (Sigma) conjugated with rabbit anti-Flagor anti-IgG antibody, and then the samples were prepared for western blot assay. For western blot, the membrane was incubated overnight at 4 °C with primary antibodies Sp1, Ets1, Flag, and Prrx1, followed by anti-mouse or -rabbit IgG (1:3000; Sigma, CA, USA), and the signals were detected with an ECL assay kit (Amersham).
For co-immunoprecipitations, the cells were lysed with 1% NP40 NET buffer (Promoter Company, China), the supernatants were incubated overnight at 4 °C with protein G-Sepharose beads (Sigma) conjugated with rabbit anti-Flagor anti-IgG antibody, and then the samples were prepared for western blot assay. For western blot, the membrane was incubated overnight at 4 °C with primary antibodies Sp1, Ets1, Flag, and Prrx1, followed by anti-mouse or -rabbit IgG (1:3000; Sigma, CA, USA), and the signals were detected with an ECL assay kit (Amersham).
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