Cells were grown to about 90% confluency in six well plates, washed twice with ice cold PBS and resuspended in modified radio‐immunoprecipitation lysis buffer (50 mM Tris/HCl, pH 7.5, 150 mM NaCl, 1% NP‐40, 0.5% Na‐desoxycholate, 1 mM dithiothreitol, 1 mM benzamidine, 1 mM PMSF). Cell suspensions were passed through a 0.45 mm needle 10 times and incubated for 15 min on ice, followed by sonication. The lysates were cleared by centrifugation at 12,000 g for 10 min at 4°C. The samples were resuspended in 5x SDS sample buffer and heated at 98°C for 5–10 min. For western blot analysis, proteins were resolved in 12% or 15% SDS polyacrylamide gels and transferred to NC membrane (GE) using the wet blot transfer over 3 hr (I = 300 mA). The blots were probed with primary antibodies, then washed with TBST (10 mM Tris/HCl, pH 8.0, 150 mM NaCl and 0.05% Tween‐20) three times and incubated with horseradish peroxidase‐conjugated secondary antibody (Thermo). The blots were washed again with TBST and signals were visualized using chemiluminescence (ECL) system (GE, Cat. No. RPN2232). The following antibodies were used: GFP mouse monoclonal antibody (proteintech, Cat. No. 66002‐1‐Ig), GAPDH mouse monoclonal antibody (proteintech, Cat. No. 60004‐1‐Ig).
Gfp mouse monoclonal antibody
Manufactured by Proteintech
The GFP mouse monoclonal antibody is a laboratory reagent used for the detection and localization of green fluorescent protein (GFP) in various applications. This antibody specifically binds to GFP, allowing for the visualization and analysis of GFP-tagged proteins or cells.
Automatically generated - may contain errors
2 protocols using gfp mouse monoclonal antibody
1
Western Blot Analysis of Protein Expression
2
Western Blot Protein Extraction and Analysis
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Cells were grown to about 90% confluency in 6 well plates, washed twice with ice cold PBS and resuspended in modified radio-immunoprecipitation (RIPA) lysis buffer (50 mM Tris/HCl, pH 7.5, 150 mM NaCl, 1% NP-40, 0.5% Na-desoxycholate, 1 mM dithiothreitol, 1 mM benzamidine, 1 mM PMSF). Cell suspensions were passed through a 0.45 μm needle 10 times and incubated for 15 min on ice, followed by sonication. The lysates were cleared by centrifugation at 10,000 rpm for 10 min at 4°C. The samples were resuspended in 5× SDS sample buffer and heated at 98°C for 5 to 10 min. For western blot analysis, proteins were resolved in 12 or 15% SDS polyacrylamide gels and transferred to NC membrane (GE) using the wet blot transfer over 3 h (I = 300 mA). The blots were probed with primary antibodies, then washed with TBST (10 mM Tris/HCl, pH 8.0, 150 mM NaCl, and 0.05% Tween-20) three times and incubated with horseradish peroxidase-conjugated secondary antibody (Thermo). The blots were washed again with TBST and signals were visualized using chemiluminescence (ECL) system (GE, Cat. NO. RPN2232). The following antibodies were used: GFP mouse monoclonal antibody (Proteintech, Cat. No. 66002-1-Ig), GAPDH mouse monoclonal antibody (Proteintech, Cat. No. 60004-1-Ig).
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