G3042

Manufactured by Merck Group

The G3042 is a laboratory equipment product offered by Merck Group. It is a general-purpose device designed for various laboratory applications. The core function of the G3042 is to [CORE FUNCTION DESCRIPTION]. Detailed specifications and intended use information is not available at this time.

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Lab products found in correlation

Ah109 and y187 haploid yeast strains, by Takara Bio (2 mentions) Gateway gene cloning technology, by Thermo Fisher Scientific (2 mentions) Horseradish peroxidase, by Bio-Rad (1 mentions) Anti myc antibody, by Roche (1 mentions) Mouse anti gfp, by Roche (1 mentions) Anti ha peroxidase high affinity, by Roche (1 mentions)

3 protocols using g3042

Immunoblot and Co-immunoprecipitation Protocols

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For immunoblot analysis, proteins were extracted from plant tissues directly into loading buffer (0.125 M Tris-HCl, pH 7.5, 4% SDS, 20% Glycerol, 0.2 M DTT, 0.02% Bromophenol Blue) while yeast protein extraction was performed following a post-alkaline method.⁵⁶ (link) For co-immunoprecipitation experiments agrobacterium cultures were infiltrated at OD600 = 0.5 and proteins were extracted as previously described.⁵⁷ (link) For immunodetection, proteins were separated by SDS-PAGE and transferred to nitrocellulose membranes which were blocked in 5% skimmed milk. Membranes were incubated with Anti-HA-Peroxidase, High Affinity (Roche, ref. 12013819001) or mouse anti-GFP (Roche, ref. 11814460001) and anti-Myc antibodies (Roche, ref. 11667149001) followed by goat anti-mouse antibodies conjugated with horseradish peroxidase (Biorad, ref. 170–5047), or polyclonal rabbit anti-luciferase (Sigma-Aldrich, REF L0159), anti-GAL4BD (Sigma-Aldrich, REF G3042) and anti-VP16 (Sigma-Aldrich, REF V4388) antibodies followed by anti-rabbit horseradish peroxidase (Sigma) to detect -HA, YFP, Myc, nLUC, cLUC, GAL4BD and VP16 tagged proteins respectively. Signals were detected using the SuperSignal West Femto chemiluminescence kit (Pierce). Ponceau S was used to stain membranes to confirm equal protein loading.

Chen J., Zhang X., Bernoux M., Rathjen J.P, & Dodds P.N. (2024). Plant Toll/interleukin-1 receptor/resistance protein domains physically associate with enhanced disease susceptibility1 family proteins in immune signaling. iScience, 27(2), 108817.

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Yeast Two-Hybrid Assay Protocol

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All plasmids used in yeast two-hybrid assays were cloned with the Gateway® Gene Cloning Technology (Invitrogen) and transformed in the AH109 and Y187 haploid yeast strains (Clontech). AH109 and Y187 cells were transformed with Gal4 DNA binding domain (GBD) fusion plasmids derived from pAS2 and Gal4 activation domain (GAD) fusion plasmids that were obtained from pGAD. Purified colonies of diploid strains were streaked on synthetic medium (SD) plates lacking leucine and tryptophan (-LW), or leucine, tryptophan and histidine (-LWH), or leucine, tryptophan and histidine with 5 mM amino-triazole (-LWH + 3AT), or leucine, tryptophan, histidine and adenine (-LWHA). Dilution assays were performed by spotting cells on -LW, -LWH, -LWH + 3AT and -LWHA plates that were incubated at 30 °C for 3 days. For verification of protein expression, protein extracts were prepared and analysed by western blotting, as previously described^{5 (link)}, with anti-GAD (1:3000; UPSTATE-06-283) and anti-GBD (1:1000; SIGMA; G3042) antibodies.

Nore A., Juarez-Martinez A.B., Clément J., Brun C., Diagouraga B., Laroussi H., Grey C., Bourbon H.M., Kadlec J., Robert T, & de Massy B. (2022). TOPOVIBL-REC114 interaction regulates meiotic DNA double-strand breaks. Nature Communications, 13, 7048.

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Yeast Two-Hybrid Assay Protocol

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All plasmids used in yeast two-hybrid assays were cloned with the Gateway® Gene Cloning Technology (Invitrogen) and transformed in the AH109 and Y187 haploid yeast strains (Clontech). AH109 and Y187 cells were transformed with Gal4 DNA binding domain (GBD) fusion plasmids derived from pAS2 and Gal4 activation domain (GAD) fusion plasmids that were obtained from pGAD. Purified colonies of diploid strains were streaked on synthetic medium (SD) plates lacking leucine and tryptophan (-LW), or leucine, tryptophan and histidine (-LWH), or leucine, tryptophan and histidine with 5 mM amino-triazole (-LWH+3AT), or leucine, tryptophan, histidine and adenine (-LWHA). Dilution assays were performed by spotting cells on -LW, -LWH, -LWH+3AT and -LWHA plates that were incubated at 30°C for 3 days. For verification of protein expression, protein extracts were prepared and analysed by western blotting, as previously described 5 , with anti-GAD (1:3000; UPSTATE-06-283) and anti-GBD (1:1000; SIGMA; G3042) antibodies.

Nore A., Juarez-Martinez A., Clément J., Brun C., Diagouraga B., Laroussi H., Grey C., Bourbon H., Kadlec J., Massy B.d., & Massy B.d. (2021). TOPOVIBL-REC114 interaction regulates meiotic DNA double-strand breaks.

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