Resolving gels were blotted to an Immobilon-FL PVDF membrane (Millipore). Membranes were blocked with Odyssey Blocking Buffer (LiCor Biosciences) diluted 1:1 with PBS and probed with either a 1:5000 dilution of rabbit 6325 antiserum for GC-A western blots or rabbit 6327 antiserum for GC-B western blots followed by 1:15,000 goat anti-rabbit IRDye 680-conjugated secondary antibody (LiCor Biosciences). FLAG western blots were probed using a 1:5000 dilution of FLAG-M2 monoclonal antibody (Sigma-Aldrich) followed by 1:15000 goat anti-mouse IRDye 800-conjugated secondary antibody (LiCor Biosciences). Actin western blots were probed using a 1:2000 dilution of a monoclonal mouse β-actin antibody (Sigma-Aldrich) followed by 1:15000 goat anti-mouse IRDye 800-conjugated secondary antibody. All western blots were visualized on a LiCor instrument as described previously (69 (link)).
Goat anti mouse irdye 800 conjugated secondary antibody
Manufactured by LI COR
The Goat anti-mouse IRDye 800-conjugated secondary antibody is a detection reagent used in western blotting, immunohistochemistry, and other immunoassays. It binds to mouse primary antibodies and emits fluorescence in the near-infrared range when excited, allowing for sensitive and quantitative detection of target proteins.
Automatically generated - may contain errors
Lab products found in correlation
Goat anti rabbit irdye 680 conjugated secondary antibody, by LI COR (2 mentions)
Odyssey imaging system, by LI COR (2 mentions)
Flag m2 monoclonal antibody, by Merck Group (1 mentions)
Immobilon fl pvdf membrane, by Merck Group (1 mentions)
Mouse monoclonal β actin antibody, by Merck Group (1 mentions)
Odyssey blocking buffer, by LI COR (1 mentions)
Ab32575, by Abcam (1 mentions)
Odyssey v3, by LI COR (1 mentions)
Sc 376021, by Santa Cruz Biotechnology (1 mentions)
Hc101 01, by Transgene (1 mentions)
Ripa lysis buffer, by Solarbio (1 mentions)
Ab8245, by Abcam (1 mentions)
3 protocols using goat anti mouse irdye 800 conjugated secondary antibody
1
Western Blot Protein Detection
2
Western Blot Analysis of BMSC Markers
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BMSCs were cultured in 10-cm tissue culture plates and treated with corresponding reagents. Radio-immunoprecipitation assay (RIPA) lysis buffer (R0010, Solarbio, Beijing, China) was used to prepare lysates. Fifty to one hundred micrograms of protein extracts were electrophoresed in 8% (for IQGAP1) or 12% (for α-SMA) SDS/polyacrylamide gels and transferred the protein to polyvinylidene fluoride (PVDF) membranes. The PVDF membranes were blocked with 5% nonfat milk at room temperature for 1 h. The blots were incubated with primary antibodies against IQGAP1 (1:1,000; sc-376021, Santa Cruz Biotechnology) or α-SMA (1:1,000; ab32575, Abcam), β-tubulin (1:5,000; HC101-01, Transgen Biotech, China), GAPDH (1:1,000; ab8245, Abcam), Cdc42 (1:1,000; 16,119, Thermo Scientific), Rac1 (1:167; 16,118, Thermo Scientific), or actin (1:2,000; Abcam). After 1 h incubation at room temperature with goat anti-rabbit IRDye-800-conjugated secondary antibody, goat anti-rabbit IRDye-700-conjugated secondary antibody, goat anti-mouse IRDye-800-conjugated secondary antibody, and goat anti-mouse IRDye-700-conjugated secondary antibody (1:10,000; 926–32,211, 926–68,021, 926–32,210, and 926–68,020, LI-COR Biosciences, Lincoln, NE), the signals were displayed using Odyssey Imaging System and quantified by the Odyssey v3.0 software (LI-COR Biosciences).
3
Western Blot Analysis of Glucocorticoid Receptor
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Tissues from control ( ) and genistein-treated ( ) mice were lysed in Tris glycine SDS sample buffer supplemented with 2-mercaptoethanol (BME). Equal amounts of protein from tissue extracts were separated via SDS-PAGE and transferred to a nitrocellulose membrane. Membranes were blocked with 7.5% skim milk in Tris-buffered saline (TBS) and incubated overnight with mouse monoclonal antibodies (1:10,000; Millipore) and rabbit monoclonal anti-GR antibodies (1:1,000; Cell Signaling Technologies) in 5% milk in TBS-Tween (0.1%) (see Table S2). Membranes were washed and incubated with goat anti-rabbit IRDye 680-conjugated secondary antibody and goat antimouse IRDye 800-conjugated secondary antibody (LI-COR Biosciences) for 1 h at room temperature. The Odyssey LI-COR imaging system (LI-COR Biosciences) was used to visualize protein expression. GR protein levels were normalized to and expressed relative to control.
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