Monoclonal α gfp antibody

Manufactured by Roche

The Monoclonal α-GFP antibody is a laboratory tool used for the detection and analysis of green fluorescent protein (GFP) in biological samples. It is a highly specific antibody that binds to GFP, allowing researchers to locate and quantify GFP-labeled proteins or cells.

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Lab products found in correlation

Image studio lite software, by LI COR (2 mentions) Ab412, by Abcam (2 mentions) α mouse horseradish peroxidase hrp antibody, by GE Healthcare (2 mentions) Pra antiserum, by Rockland Immunochemicals (2 mentions) Anti rabbit igg h l hrpo, by Dianova (2 mentions) Anti mouse igg h l hrpo, by Dianova (2 mentions) Fusion sl chemiluminescence detection system, by Avantor (2 mentions) Amersham ecl prime western blotting detection reagent, by GE Healthcare (2 mentions) Flag antibody, by Merck Group (1 mentions) Bca protein assay kit, by CWBIO (1 mentions)

3 protocols using monoclonal α gfp antibody

Protein Expression and Detection Protocol

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GFP-tagged proteins were recognized with a monoclonal α-GFP antibody (Roche; 1:1000) and arginine methylation was recognized with a monoclonal α-mono and dimethyl arginine antibody (Ab412, Abcam; 1:500), followed by a secondary α-mouse-horseradish peroxidase (HRP) antibody (GE Lifesciences; 1:5000). Slr1-TAP was recognized with an HRP-conjugated goat anti-Protein A (PrA) antiserum (Rockland; 1:10,000). Ribosomal protein Rps3 was recognized with a polyclonal rabbit antibody against Rps3 (1:1000). Proteins were visualized through enhanced chemiluminescence (Pierce) and autoradiography.
Anti-rabbit IgG (H+L)-HRPO and anti-mouse IgG (H+L) HRPO (Dianova) secondary antibodies were used for sucrose density gradient fraction immunoblots. The signals were detected with Amersham ECL Prime Western Blotting Detection Reagent (GE Healthcare) and the FUSION-SL chemiluminescence detection system (Peqlab) and the Western blot analyses were quantified using the Image StudioLite software (LI-COR).

Ariyachet C., Beißel C., Li X., Lorrey S., Mackenzie O., Martin P.M., O'Brien K., Pholcharee T., Sim S., Krebber H, & McBride A.E. (2017). Post‐translational modification directs nuclear and hyphal tip localization of C andida albicans mRNA‐binding protein Slr1. Molecular Microbiology, 104(3), 499-519.

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Immunoblotting and Quantification of Methylated Proteins

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GFP‐tagged proteins were recognized with a monoclonal α‐GFP antibody (Roche; 1:1000) and arginine methylation was recognized with a monoclonal α‐mono and dimethyl arginine antibody (Ab412, Abcam; 1:500), followed by a secondary α‐mouse‐horseradish peroxidase (HRP) antibody (GE Lifesciences; 1:5000). Slr1‐TAP was recognized with an HRP‐conjugated goat anti‐Protein A (PrA) antiserum (Rockland; 1:10,000). Ribosomal protein Rps3 was recognized with a polyclonal rabbit antibody against Rps3 (1:1000). Proteins were visualized through enhanced chemiluminescence (Pierce) and autoradiography.
Anti‐rabbit IgG (H + L)‐HRPO and anti‐mouse IgG (H + L) HRPO (Dianova) secondary antibodies were used for sucrose density gradient fraction immunoblots. The signals were detected with Amersham ECL Prime Western Blotting Detection Reagent (GE Healthcare) and the FUSION‐SL chemiluminescence detection system (Peqlab) and the Western blot analyses were quantified using the Image StudioLite software (LI‐COR).

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Optimized Protein Extraction and Western Blot Analysis

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The leaves were excised, flash-frozen in liquid nitrogen, and pulverized. Total protein was extracted with denaturing buffer [4 M urea, 65 mM Tris–HCl (pH 7.3), 3% (v/v) sodium dodecyl sulfate (SDS), 10% (v/v) glycerol, 0.05% (w/v) bromophenol blue, and 10 mM dithiothreitol (DTT)]. The protein concentrations in the supernatants were determined with a bicinchoninic acid (BCA) protein assay kit (CW0014; CWBiotech, Beijing, China). The proteins were loaded onto 10% SDS-PAGE and transferred to polyvinylidene fluoride (PVDF) membranes which were then incubated with a monoclonal α-GFP antibody (Roche; Cat-No: 11814460001, monoclonal, mouse, dilution 1:2000) and a FLAG antibody (Sigma-Aldrich; Cat-No: F4042, monoclonal, mouse, dilution 1:3000). Anti-mouse horseradish peroxidase (HRP) secondary antibody (Abcam, Cat-No: ab205719, dilution 1:10,000) was used for detection. The PVDF membranes were then incubated in Ponceau S solution [40% (v/v) methanol, 15% (v/v) acetic acid, and 0.25% (w/v) Ponceau S] and destained with deionized water. Images were captured with ChemiDoc^TM XRS 170-8070 (Bio-Rad Laboratories, Hercules, CA, United States).

Xu T., Wang X., Ma H., Su L., Wang W., Meng J, & Xu Y. (2021). Functional Characterization of VDACs in Grape and Its Putative Role in Response to Pathogen Stress. Frontiers in Plant Science, 12, 670505.

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