Separation of proteins were performed by SDS-PAGE using a Mini PROTEAN system (Bio-Rad) or Novex NuPAGE SDS-PAGE Gel system (Life technologies) and transferred onto Roti-PVDF membranes (0.45μm, Roth) using a Mini TransBlot Electrophoretic Transfer Cell device (Bio-Rad). Precision Plus Dual Color Protein Standard (Bio-Rad) was used as a marker. Membranes were blocked with 4% (w/v) milk (Roth) in TBST (50 mM Tris, 150 mM NaCl, pH 7.2, 0.1% (v/v) Tween 20) for 30 min at room temperature. Binding of primary antibodies was carried out either at 4°C over night or for 1h at room temperature. Suitable secondary antibodies (coupled to horseradish peroxidase, Dianova) were incubated for 30min at room temperature. All antibodies were diluted in blocking medium. Bands were analyzed densitometrically using ImageLab Software (Biorad) and MBP Signals were normalized to CNP Signals.
Mini transblot electrophoretic transfer cell device
The Mini TransBlot Electrophoretic Transfer Cell device is a laboratory equipment used for the transfer of proteins or nucleic acids from a gel to a membrane for further analysis. It provides a controlled environment for the efficient transfer of these biological molecules.
Lab products found in correlation
3 protocols using mini transblot electrophoretic transfer cell device
Protein Extraction and Analysis from Mouse Sciatic Nerve and Brain
Separation of proteins were performed by SDS-PAGE using a Mini PROTEAN system (Bio-Rad) or Novex NuPAGE SDS-PAGE Gel system (Life technologies) and transferred onto Roti-PVDF membranes (0.45μm, Roth) using a Mini TransBlot Electrophoretic Transfer Cell device (Bio-Rad). Precision Plus Dual Color Protein Standard (Bio-Rad) was used as a marker. Membranes were blocked with 4% (w/v) milk (Roth) in TBST (50 mM Tris, 150 mM NaCl, pH 7.2, 0.1% (v/v) Tween 20) for 30 min at room temperature. Binding of primary antibodies was carried out either at 4°C over night or for 1h at room temperature. Suitable secondary antibodies (coupled to horseradish peroxidase, Dianova) were incubated for 30min at room temperature. All antibodies were diluted in blocking medium. Bands were analyzed densitometrically using ImageLab Software (Biorad) and MBP Signals were normalized to CNP Signals.
Western Blot Protein Analysis Protocol
Proteins were separated by SDS-PAGE using a Mini PROTEAN 3 system (Bio-Rad) and transferred onto PVDF membranes (Immobilion-P, Millipore) using a Mini TransBlot Electrophoretic Transfer Cell device (Bio-Rad). To block unspecific binding, membranes were incubated with 4% (w/v) milk (Roth) in TBST (0.05 M Tris, 0.15 M NaCl, pH 7.2, 0.1% (v/v) Tween20) for 30 min at room temperature. Primary antibody binding was carried out overnight at 4°C and appropriate secondary antibodies (coupled with horseradish peroxidase, Dianova) were incubated for 30 min at room temperature, both in blocking medium.
Western Blot Protein Extraction and Analysis
Separation of proteins was performed by SDS-PAGE using a Mini PROTEAN system (Bio-Rad) or Novex NuPAGE SDS-PAGE Gel system (Life Technologies) and transferred onto Roti-PVDF membranes (0.45 μm, Roth) using a Mini TransBlot Electrophoretic Transfer Cell device (Bio-Rad). Precision Plus Protein Standard (Bio-Rad) was used as a marker. Membranes were blocked with 4% (w/v) milk (Roth) in TBST [50 mM Tris, 150 mM NaCl, pH 7.2, 0,1% (v/v) Tween 20] for 30 min at room temperature. Binding of primary antibodies was carried out either at 4°C over night or for 1 h at room temperature. Suitable secondary antibodies (coupled to horseradish peroxidase, Dianova) were incubated for 30 min at room temperature. All antibodies were diluted in blocking medium. Image acquisition was performed in a ChemiDoc XRS system using Imagelab software (Biorad).
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