Western blot analysis was performed as described previously [3 (link),6 (link)]. Hippocampal tissues were homogenized with lysis buffer. Protein content was measured using a Bio-Rad colorimetric protein assay kit (Bio-Rad, Hercules, CA, USA). The protein mixture containing 40 μg total protein was separated on sodium dodecyl sulfate-polyacrylamide gels and transferred onto a nitrocellulose membrane. Mouse actin antibody (1:2,000; Santa Cruz Biotechnology), mouse Bax antibody (1:1,000; Santa Cruz Biotechnology) and mouse Bcl-2 antibody (1:1,000; Santa Cruz Biotechnology) were used as primary antibodies. Horseradish peroxidase-conjugated antimouse antibodies for Bax and Bcl-2 (1:2,000; Amersham Pharmacia Biothech GmbH, Freiburg, Germany) were used as secondary antibodies. Bands were detected using an enhanced chemiluminescence detection kit (Santa Cruz Biotechnology).
Mouse bax antibody
Manufactured by Santa Cruz Biotechnology
Sourced in United States
The Mouse Bax antibody is a research-use-only product designed to detect the Bax protein, a member of the Bcl-2 family, in mouse samples. This antibody can be used in various immunoassay applications to study the role of Bax in biological processes.
Automatically generated - may contain errors
Lab products found in correlation
Mouse bcl 2 antibody, by Santa Cruz Biotechnology (4 mentions)
Colorimetric protein assay kit, by Bio-Rad (2 mentions)
Horseradish peroxidase conjugated anti mouse antibody, by Vector Laboratories (2 mentions)
Enhanced chemiluminescence detection kit, by Santa Cruz Biotechnology (1 mentions)
Mouse β actin antibody, by Santa Cruz Biotechnology (1 mentions)
Molecular analyst version 1, by Bio-Rad (1 mentions)
Horseradish peroxidase conjugated anti rabbit antibody, by Vector Laboratories (1 mentions)
β actin antibody, by Santa Cruz Biotechnology (1 mentions)
Ecl detection kit, by Santa Cruz Biotechnology (1 mentions)
Mouse beta actin antibody, by Santa Cruz Biotechnology (1 mentions)
4 protocols using mouse bax antibody
1
Hippocampal Protein Expression Analysis
2
Western Blot Analysis of Achilles Tendon Proteins
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Western blot analysis was done, like the method described below [20 (link),21 (link)]. The Achilles tendon tissues were homogenized with lysis buffer containing 50mM Tris-HCl (pH, 8.0), 100mM NaF, 150mM NaCl, 1.5mM MgCl2·6H2O, 1mM EGTA, 1mM PMSF, 1mM Na2VO4, 10% glycerol, 1% Triton X-100, and then centrifuged for 30 minutes at 14,000 rpm. Rabbit CREB antibody (1:1,000; Abcam), rabbit phosphorylated CREB (p-CREB) antibody (1:1,000; Santa Cruz Biotechnology, Santa Cruz, CA, USA), rabbit PKA antibody (1:1,000; Santa Cruz Biotechnology), rabbit phosphorylated PKA (p-PKA) antibody (1:1,000; Santa Cruz Biotechnology), mouse Bax antibody (1:1,000; Santa Cruz Biotechnology), mouse Bcl-2 antibody (1:1,000; Santa Cruz Biotechnology), and mouse β-actin antibody (1:1,000; Santa Cruz Biotechnology) were chosen as the primary antibodies. Horseradish peroxidase-conjugated anti-mouse antibody (1:2,000; Vector Laboratories) for β-actin, Bax, Bcl-2, and anti-rabbit antibody (1:2000; Vector Laboratories) for CREB, pCREB, PKA, p-PKA were chosen as the secondary antibodies. Membrane transfer was carried out at 4°C, and all other steps were conducted at room temperature. The bands were calculated by Molecular Analyst version 1.4.1 (Bio-Rad Laboratories, Hercules, CA, USA).
3
Protein Expression Analysis in Hippocampal Tissue
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Western blot analysis for NF-κB, IκB-α, Bax, Bcl-2, MMP-9 expression was used as explained below (Ko et al., 2020 (link); Park et al., 2020 (link)). Hippocampal tissues were lysed in a lysis buffer containing 150 mM NaCl, 50 mM Tris-HCl (pH, 7.5), 1 mM phenylmethylsulfonyl fluoride, 100-mg/mL leupeptin, 1% Nonidet P40, 0.5% deoxycholic acid, 0.1% sodium dodecyl sulfate. Rabbit NF-κB antibody (1:1,000; Abcam), rabbit IκB-α antibody, rabbit phosphorylated IκB-α (p-IκB-α) antibody (1:1,000; Santa Cruz Biotechnology, Santa Cruz, CA, USA), rabbit MMP antibody (1:2,000; Cell Signaling Technology, Inc., Danvers, USA), mouse Bax antibody, mouse Bcl-2 antibody, and β-actin antibody (1:1,000; Santa Cruz Biotechnology) were used as the primary antibodies. Horseradish peroxidase-conjugated anti-mouse antibody (1:2,000; Vector Laboratories, Burlingame, CA, USA) for β-actin, Bax, Bcl-2, and horseradish peroxidase-conjugated anti-rabbit antibody (1: 2,000; Vector Laboratories) for NF-κB, IκB-α, and MMP were used as the secondary antibodies. Image-Pro Plus computer-aided image analysis system (Media Cybernetics Inc., Silver Spring, MD, USA) was used for band quantification.
4
Western Blot Analysis of Hippocampal Proteins
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The hippocampal tissues were collected, and then were immediately frozen at −70°C. The hippocampal tissues were homogenized on ice, and lysed in a lysis buffer containing 50 mM HEPES (pH 7.5), 150 mM NaCl, 10% glycerol, 1% Triton X-100, 1 mM PMSF, 1 mM EGTA, 1.5 mM MgCl2 6H2O, 1 mM sodium orthovanadate, and 100 mM sodium flouride. Protein content was measured using a Bio-Rad colorimetric protein assay kit (Bio-Rad, Hercules, CA, USA). Protein (30 μg) was separated on SDS-polyacrylamide gels and transferred onto a nitrocellulose membrane.
Mouse beta-actin antibody (1:500; Santa Cruz Biotechnology), mouse Bcl-2 antibody (1:1,000; Santa Cruz Biotechnology), and mouse Bax antibody (1:1,000; Santa Cruz Biotechnology) were used as the primary antibodies. Horseradish peroxidase-conjugated anti-mouse antibodies for beat-actin, Bax, and Bcl-2 (1:3,000; Amersham Pharmacia Biothech GmbH, Freiburg, Germany) were used as the secondary antibodies.
Experiment was performed in normal lab conditions and at room temperature except membrane transfer. Membrane transfer was performed at 4ºC with the cold pack and pre-chilled buffer. Band detection was performed using the enhanced chemiluminescence (ECL) detection kit (Santa Cruz Biotechnology).
Mouse beta-actin antibody (1:500; Santa Cruz Biotechnology), mouse Bcl-2 antibody (1:1,000; Santa Cruz Biotechnology), and mouse Bax antibody (1:1,000; Santa Cruz Biotechnology) were used as the primary antibodies. Horseradish peroxidase-conjugated anti-mouse antibodies for beat-actin, Bax, and Bcl-2 (1:3,000; Amersham Pharmacia Biothech GmbH, Freiburg, Germany) were used as the secondary antibodies.
Experiment was performed in normal lab conditions and at room temperature except membrane transfer. Membrane transfer was performed at 4ºC with the cold pack and pre-chilled buffer. Band detection was performed using the enhanced chemiluminescence (ECL) detection kit (Santa Cruz Biotechnology).
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