Frozen muscle tissue or spinal cord (L6-S1) tissue was homogenized in ice-cold lysis buffer (Applygen Technologies Inc., Beijing, China) with protease inhibitor cocktail (Roche, Switzerland). Equal amounts (50 μg) of extract were separated by 10% SDS-PAGE and transferred to a polyvinylidene fluoride membrane (Millipore, United States) at 4°C. Non-specific binding sites were blocked with 10% non-fat milk in TBS (20 mM Tris-Cl, pH 7.5, containing 0.15 M NaCl and 2.7 mM KCl) for 1 h at room temperature. The membrane was incubated with the primary antibodies listed in Table 1 overnight at 4°C. After washes with TBST, the membrane was incubated with the appropriate secondary antibodies goat anti-rabbit IgG (1:10,000, 611-132-122, Rockland, United States) or sheep anti-mouse IgG (1:10,000, 610-632-002, Rockland, United States) for 2 h at room temperature. The membrane was washed in TBST and visualized with an Odyssey Infrared Imager (LI-COR, NE, United States). The integrated intensity of the bands was analyzed with Odyssey software (version 1.2).
Sheep anti mouse igg
Manufactured by Rockland Immunochemicals
Sourced in United States, China
Sheep anti-mouse IgG is a secondary antibody used in various immunoassays and research applications to detect the presence of mouse immunoglobulin G (IgG) in samples. It is produced by immunizing sheep with mouse IgG, and the resulting antibodies are purified and conjugated as needed for specific applications.
Automatically generated - may contain errors
Lab products found in correlation
Polyvinylidene fluoride membrane, by Merck Group (1 mentions)
Goat anti rabbit igg, by Rockland Immunochemicals (1 mentions)
Odyssey infrared imager, by LI COR (1 mentions)
Protease inhibitor cocktail, by Roche (1 mentions)
Lysis buffer, by Applygen (1 mentions)
Dm lb2, by Leica (1 mentions)
Donkey anti mouse igg, by Thermo Fisher Scientific (1 mentions)
Goat serum, by Merck Group (1 mentions)
2 protocols using sheep anti mouse igg
1
Western Blot Analysis of Muscle and Spinal Cord
2
Immunohistochemical Analysis of Dopaminergic Neurons
Check if the same lab product or an alternative is used in the 5 most similar protocols
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The brain slices were fixed with cold acetone for 15 min, and then washed (3×5 min) in 0.3 % Triton X-100 phosphate buffer solution (PBST) to eliminate the residual fixative. After blocking with 3 % H 2 O 2 and 10 % goat serum (Sigma-Aldrich, St. Louis, MO, USA) at room temperature for 30 min, the sections were incubated with TH antibody (Mouse, 1:10000, Sigma/T1299) at 4 ℃ overnight. After washing in PBST (3×5 min), sections of the SN were incubated with donkey antimouse IgG (1:1000, Invitrogen/A21203) for 1 h at room temperature and then observed under a fluorescence microscope (Leica DM LB2, St. Gallen, Switzerland). The sections of the striatum were incubated with sheep anti-mouse IgG (1:1000, Rockland/13175) for 1 h at room temperature and incubated with 3, 3'-diaminobenzidine tetrahydrochloride (DAB Substrate Kit for Peroxidase, Beyotime Biotechnology, Shanghai, China) for 2 min, stopped with water, and then placed in xylene and overlaid with a coverslip using neutral resin-mounting medium.
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