The frozen kidney of each rat was thawed, dissected into the cortex and medulla, and then homogenized in 100 mmol·L−1 potassium buffer (pH 7.25) containing 30% glycerol, 1 mmol·L−1 dithiothreitol, and 0.1 mmol·L−1 phenylmethylsulfonyl fluoride (15 (link)). Protein expression and phosphorylation were examined using Western blot analysis, as described previously (19 (link)). Antibodies against Raf-B (no. 5284, Santa Cruz), ERK (no. 4695, Cell Signaling Technology), p-ERK (no. 4376, Cell Signaling Technology), mTOR (no. 2983, Cell Signaling Technology), p-mTOR (no. 2971, Cell Signaling Technology), S6 (no. 2217, Cell Signaling Technology), and p-S6 (no. 2211, Cell Signaling Technology) were used. Secondary HRP-conjugated mouse antirabbit (no. 2357, Santa Cruz) and rabbit antimouse (no. 516102, Santa Cruz) antibodies were then used. Relative band intensities were quantified using ImageJ and normalized using β-actin (A2228; Sigma-Aldrich, St. Louis, MO) as an internal standard.
Rabbit antimouse
Manufactured by Santa Cruz Biotechnology
Rabbit antimouse is a reagent used in immunoassays and other laboratory techniques. It is a polyclonal antibody produced in rabbits that specifically recognizes and binds to mouse proteins or antigens. The core function of this product is to facilitate the detection and analysis of mouse-derived samples in various experimental settings.
Automatically generated - may contain errors
2 protocols using rabbit antimouse
1
Quantitative Analysis of Protein Expression
2
Kidney Protein Extraction and Analysis
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The frozen kidney of each rat was thawed, dissected into the cortex and medulla, and then homogenized in 100 mmol/L potassium buffer (pH 7.25) containing 30% glycerol, 1 mmol/L (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission.
dithiothreitol, and 0.1 mmol/L phenylmethylsulfonyl fluoride (14). Protein expression and phosphorylation were examined using western blot analysis, as described previously (18) .
Antibodies against Raf-B (#5284; Santa Cruz), ERK (#4695; Cell Signaling Technology), p-ERK (#4376; Cell Signaling Technology), mTOR (#2983; Cell Signaling Technology), p-mTOR (#2971; Cell Signaling Technology), S6 (#2217; Cell Signaling Technology), and p-S6 (#2211; Cell Signaling Technology) were used. Secondary HRP-conjugated mouse anti-rabbit (#2357; Santa Cruz) and rabbit anti-mouse (#516102; Santa Cruz) antibodies were then used. Relative band intensities were quantified using ImageJ and normalized using β-actin (A2228; Sigma-Aldrich, St.
Louis, MO, USA) as an internal standard.
dithiothreitol, and 0.1 mmol/L phenylmethylsulfonyl fluoride (14). Protein expression and phosphorylation were examined using western blot analysis, as described previously (18) .
Antibodies against Raf-B (#5284; Santa Cruz), ERK (#4695; Cell Signaling Technology), p-ERK (#4376; Cell Signaling Technology), mTOR (#2983; Cell Signaling Technology), p-mTOR (#2971; Cell Signaling Technology), S6 (#2217; Cell Signaling Technology), and p-S6 (#2211; Cell Signaling Technology) were used. Secondary HRP-conjugated mouse anti-rabbit (#2357; Santa Cruz) and rabbit anti-mouse (#516102; Santa Cruz) antibodies were then used. Relative band intensities were quantified using ImageJ and normalized using β-actin (A2228; Sigma-Aldrich, St.
Louis, MO, USA) as an internal standard.
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