α 33p gtp

Manufactured by PerkinElmer

α-33P GTP is a radioactive nucleotide used for labeling and detection in various molecular biology applications. It provides a stable and high-energy phosphate label for studies involving DNA, RNA, or protein synthesis and interactions.

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Lab products found in correlation

Typhoon trio, by GE Healthcare (1 mentions) Adenosine triphosphate (atp), by PerkinElmer (1 mentions) Superase in, by Thermo Fisher Scientific (1 mentions) Dna and rna oligonucleotides, by Integrated DNA Technologies (1 mentions) Ambion t7 megashortscript kit, by Thermo Fisher Scientific (1 mentions) Proteinase k, by Thermo Fisher Scientific (1 mentions) γ 33p atp, by PerkinElmer (1 mentions) Hiv 1 rt, by Worthington (1 mentions)

2 protocols using α 33p gtp

Nucleotide Analog Inhibition Assay

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Nucleotide analog inhibitors were serially diluted 3-fold in water and added to reaction mixture containing 10% virus cRNP (v/v), 100 mM Tris-HCl (pH 8.0), 100 mM KCl, 1 mM DTT, 10% glycerol, 0.25% Triton-101 (reduced), 5 mM MgCl₂, 0.4 U/mL RNasin, and 200 μM ApG dinucleotide primer (TriLink, San Diego, CA). Reactions were initiated by addition of ribonucleotide triphosphate (NTP) substrate mix containing 1 μM GTP (at 0.5 x K_m), 0.01 μM α-³³P GTP, and 100 μM of ATP, CTP, and UTP (PerkinElmer, Shelton, CT). Reactions were incubated at 30°C for 90 minutes then spotted onto DE81 filter paper. Filters were air dried, washed with 0.125 M Na₂HPO4 (3×), water (1×), and Ethanol (1×) and air dried before exposure to Typhoon phosphor imager screen. Product formation was quantified on a Typhoon Trio (GE Healthcare, Piscataway, NJ). IC₅₀ values were calculated for inhibitors by fitting the data to a four-parameter sigmoidal dose response with variable slope in GraphPad Prism.

Barauskas O., Xing W., Aguayo E., Willkom M., Sapre A., Clarke M., Birkus G., Schultz B.E., Sakowicz R., Kwon H, & Feng J.Y. (2017). Biochemical characterization of recombinant influenza A polymerase heterotrimer complex: Polymerase activity and mechanisms of action of nucleotide analogs. PLoS ONE, 12(10), e0185998.

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Preparation of Radiolabeled HIV-1 Nucleic Acid Substrates

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DNA and RNA oligonucleotides were obtained from Integrated DNA Technologies (IDT) (Coralville, IA). T4 polynucleotide kinase, proteinase K, SUPERaseIN, and Gel Loading Buffer II were purchased from Applied Biosystems (now Life Technologies) (Foster City, CA). [γ-³³P]ATP, [α-³³P]dCTP, and [α-³³P]GTP were purchased from PerkinElmer (Shelton, CT). Purified tRNA^Lys3 from human placenta was obtained from Bio S&T (Lachine, Québec, Canada). RNA transcripts were synthesized by using the Ambion MEGAshortscript T7 kit (Life Technologies). HIV-1 RT was obtained from Worthington (Lakewood, NJ). The SP2 peptide (FLGKIWPSHKGRPGNF) was purchased from GenScript (Piscataway, NJ). The sequences of the viral nucleic acid substrates and the RNA 200 and RNA 60 templates were derived from HIV-1 NL4-3 (GenBank Accession no. AF324493) (Adachi et al., 1986 (link)). The RNA 105 template was derived from the HIV-1 HXB2 sequence (GenBank Accession no. K03455), as described previously (Hargittai et al., 2001 (link)).

Wu T., Gorelick R.J, & Levin J.G. (2014). Selection of fully processed HIV-1 nucleocapsid protein is required for optimal nucleic acid chaperone activity in reverse transcription. Virus research, 193, 52-64.

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