M7gdp

Manufactured by Merck Group

The M7GDP is a laboratory equipment product from Merck Group. It is designed for performing basic analytical tasks in research and development settings. The core function of the M7GDP is to facilitate the measurement and analysis of various samples and substances. For a more detailed and unbiased description, please consult the product specifications or technical documentation.

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Lab products found in correlation

Anti eif4e, by BD (1 mentions) Adipic acid dihydrazide agarose, by Merck Group (1 mentions)

2 protocols using m7gdp

Analyzing LRP130 and Importin 8 Interactions

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Antibodies used were rabbit polyclonal anti-LRP130 (H-300; Santa Cruz Bio.), rabbit polyclonal anti-Importin 8 (LifeSpan BioSciences), mouse monoclonal anti-eIF4E (BD Transduction Laboratories), goat polyclonal anti-GST (GE Health Sciences), and anti-goat Cy3 (Jackson ImmunoResearch). The cap analog m⁷GDP was obtained from Sigma. The synthetic peptide LRP10A (N-Ac-EGFPIRPHYFWPLLVGRRKEK-NH₂), in >95% purity was purchased from Biomatic Corporation and stored as a lyophilized powder. The RNA was purchased from Dharmacon, Inc. Prior to use, each nucleotide was heated at 85°C for 5 min, followed by a ramp to 60°C at 0.1°C/sec, 37°C for 5 min, then 25°C for 20 min.

Volpon L., Culjkovic-Kraljacic B., Sohn H.S., Blanchet-Cohen A., Osborne M.J, & Borden K.L. (2017). A biochemical framework for eIF4E-dependent mRNA export and nuclear recycling of the export machinery. RNA, 23(6), 927-937.

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Synthesis and Immobilization of m7GxP

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The synthesis was adapted from that of Edery et al. [52 (link)]. m⁷GMP (synthesized; see below) or m⁷GDP (Sigma) sodium salts was dissolved in water (10 mM final), and a solution of sodium periodate (1.0 equiv) in sodium acetate buffer (0.1 M, pH 6; 51 mM final) was added. The resulting mixture was agitated at 25 °C for 30 min protected from light. Adipic acid dihydrazide agarose (1 mL packed; Sigma) was washed with water (1× 20 mL) followed by sodium acetate buffer (1× 20mL), and then resuspended in sodium acetate buffer (2 mL). To this slurry was added aniline (20 equiv) and the oxidized m⁷GxP solution. The resin mixture was then shaken at 25 °C for 45 min before adding sodium cyanoborohydride (16 equiv). The mixture was agitated overnight at 4 °C. The resin was then washed with NaCl (1M; 5× 5 mL), equilibrated in buffer A (5 mL; 50 mM HEPES, pH 7, 200 mM KCl), and stored at 4 °C.

Kaur T., Menon A, & Garner A.L. (2019). Synthesis of 7-Benzylguanosine Cap Analogue Conjugates for eIF4E Targeted Degradation. European journal of medicinal chemistry, 166, 339-350.

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