Soluble whole-cell lysates of the hMBs treated with 30 μM of iSN04 or AS1411 in DM for 48 h were prepared as described above. The lysates were denatured with 50 mM Tris-HCl (pH 6.8), 10% glycerol, and 2% SDS at 95°C for 5 min. Ten microgram of protein samples were subjected to SDS-PAGE on a 10% polyacrylamide gel followed by Western blotting using an iBlot 2 Dry Blotting System (Thermo Fisher Scientific). 1.0 μg/ml each of rabbit polyclonal anti-nucleolin antibody, mouse monoclonal anti-p53 antibody (PAb 240; Abcam), and mouse monoclonal anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) antibody (5A12; Wako) were used as primary antibodies. 0.1 μg/ml each of horseradish peroxidase (HRP)-conjugated goat anti-rabbit and anti-mouse IgG antibodies (Jackson ImmunoResearch) were used as secondary antibodies, respectively. HRP activity was detected using ECL Prime reagents (GE Healthcare) and ImageQuant LAS 500. The quantities of nucleolin and p53 proteins were normalized to that of GAPDH using ImageJ software.
Horseradish peroxidase hrp conjugated goat anti rabbit and anti mouse igg antibodies
Manufactured by Jackson ImmunoResearch
Horseradish peroxidase (HRP)-conjugated goat anti-rabbit and anti-mouse IgG antibodies are laboratory reagents used for detection and quantification purposes in various immunoassays and immunochemical techniques. These antibodies are conjugated with the enzyme horseradish peroxidase, which catalyzes a colorimetric or chemiluminescent reaction when exposed to appropriate substrates, allowing for the visualization and measurement of target analytes.
Automatically generated - may contain errors
Lab products found in correlation
Iblot 2 dry blotting system, by Thermo Fisher Scientific (2 mentions)
Pab240, by Abcam (2 mentions)
Ecl prime reagent, by GE Healthcare (1 mentions)
Imagequant las 500, by GE Healthcare (1 mentions)
Gapdh antibody, by Fujifilm (1 mentions)
Cycloheximide (chx), by Merck Group (1 mentions)
Ara c, by Merck Group (1 mentions)
β actin, by Thermo Fisher Scientific (1 mentions)
3 protocols using horseradish peroxidase hrp conjugated goat anti rabbit and anti mouse igg antibodies
1
Quantitative Analysis of Nucleolin and p53
2
Antibodies for Investigating Cellular Pathways
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Mouse monoclonal antibody for DHX9, G3BP1, and goat polyclonal antibody for TIA1 were purchased from Santa Cruz Biotechnology. Rabbit polyclonal antibody for DHX9, mouse antibody for V5, and β-actin were purchased from Thermo Fisher Scientific. Cycloheximide and AraC were purchased from Sigma. Mouse monoclonal antibody for dsRNA SCICONS J2 was from English and Scientific Consulting Kft., Hungary (SCICONS). Horseradish peroxidase (HRP)-conjugated goat anti-rabbit and anti-mouse IgG antibodies were purchased from Jackson ImmunoResearch Laboratories. All the secondary antibodies conjugated to Alexa Fluor 488, 594, 568, and 647 were purchased from Thermo Fisher Scientific.
3
Nucleolin and p53 Protein Analysis
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Soluble whole-cell lysates of the hMBs treated with 30 μM of iSN04 or AS1411 in DM for 48 h were prepared as described above. The lysates were denatured with 50 mM Tris-HCl, 10% glycerol, and 2% SDS at 95°C for 5 min.
10 μg of protein samples were subjected to SDS-PAGE on a 10% polyacrylamide gel followed by Western blotting using an iBlot 2 Dry Blotting System (Thermo Fisher Scientific). 1.0 μg/ml each of rabbit polyclonal antinucleolin antibody, mouse monoclonal anti-p53 antibody (PAb 240; Abcam), and mouse monoclonal anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) antibody (5A12; Wako) were used as primary antibodies. 0.1 μg/ml each of horseradish peroxidase (HRP)-conjugated goat anti-rabbit and antimouse IgG antibodies (Jackson ImmunoResearch) were used as secondary antibodies, respectively. HRP activity was detected using ECL Prime reagents and ImageQuant LAS 500. The quantities of nucleolin and p53 proteins were normalized to that of GAPDH using ImageJ software.
10 μg of protein samples were subjected to SDS-PAGE on a 10% polyacrylamide gel followed by Western blotting using an iBlot 2 Dry Blotting System (Thermo Fisher Scientific). 1.0 μg/ml each of rabbit polyclonal antinucleolin antibody, mouse monoclonal anti-p53 antibody (PAb 240; Abcam), and mouse monoclonal anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) antibody (5A12; Wako) were used as primary antibodies. 0.1 μg/ml each of horseradish peroxidase (HRP)-conjugated goat anti-rabbit and antimouse IgG antibodies (Jackson ImmunoResearch) were used as secondary antibodies, respectively. HRP activity was detected using ECL Prime reagents and ImageQuant LAS 500. The quantities of nucleolin and p53 proteins were normalized to that of GAPDH using ImageJ software.
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