Inactivation buffer

Manufactured by Thermo Fisher Scientific

Inactivation buffer is a laboratory solution designed to inactivate or neutralize biological samples, such as enzymes, viruses, or other potentially hazardous materials. Its core function is to provide a controlled environment for the safe handling and processing of these samples.

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Lab products found in correlation

End labeled decade marker, by Thermo Fisher Scientific (2 mentions) γ 32 atp, by PerkinElmer (2 mentions) Rpa 3 kit, by Thermo Fisher Scientific (1 mentions) Decade marker, by Thermo Fisher Scientific (1 mentions) Rnase t1, by Thermo Fisher Scientific (1 mentions) Rnase v1, by Thermo Fisher Scientific (1 mentions) Rnase a, by Thermo Fisher Scientific (1 mentions) Ribonuclease protection assay 3 kit, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

Inactivation/precipitation buffer (5 citations)

4 protocols using inactivation buffer

Ribonuclease Protection Assay for HvCESA1

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Ribonuclease protection assays (RPAs) were performed by using the RPA III kit (Ambion) according to manufacturer's instructions and as previously described (Held et al., 2008 (link)). Labeled sense and antisense riboprobes targeting HvCESA1 and control probes were gel purified by 5% PAGE containing 8 M urea in 1× TBE buffer per kit instructions and hybridized with 10‐ to 20‐μg total RNA from either barley, yeast, or mouse for 16–18 h at 42°C. Reaction mixtures were digested with RNase A/T1 (1:100) for 30 min at 37°C and then stopped with inactivation buffer (Ambion), and protected fragments were precipitated by using 10‐μg yeast RNA as a carrier. The protected fragments were separated by 12.5% PAGE containing 8 M urea in 1× TBE buffer. γ‐³²ATP (Perkin Elmer Health Sciences) end‐labeled Decade Marker (Ambion), prepared per manufacturer's protocol, served as the size standard. Autoradiograms of RPA gels were uniformly scanned at 600‐dpi grayscale in a lossless format. The intensity of bands in the 21‐ to 24‐nt range were analyzed using ImageJ (Version 1.49E).

Nething D.B., Sukul A., Mishler‐Elmore J.W, & Held M.A. (2021). Posttranscriptional regulation of cellulose synthase genes by small RNAs derived from cellulose synthase antisense transcripts. Plant Direct, 5(9), e347.

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FBP1 Binding to EV71 5' UTR

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The 5' end-labeled EV71 5' UTR linker region was incubated at 4°C for 10 minutes with 2 μg of full-length FBP1 or 1.14 μg of FBP1^1-371 in binding buffer [57 (link)] containing 1 μl of 0.02 μg/μl RNase A or 0.02 U/ml RNase T1. The reactions were terminated with 10 μl of inactivation buffer (Ambion, Austin, TX). The cleavage products were separated in 12% acrylamide/7M urea gels, after which the gels were dried and subjected to a phosphor image scan. Nucleotide positions were determined through comparison with the Decade marker (Ambion). Footprinting experiments were performed in triplicate, with similar results derived overall.

Hung C.T., Kung Y.A., Li M.L., Brewer G., Lee K.M., Liu S.T, & Shih S.R. (2016). Additive Promotion of Viral Internal Ribosome Entry Site-Mediated Translation by Far Upstream Element-Binding Protein 1 and an Enterovirus 71-Induced Cleavage Product. PLoS Pathogens, 12(10), e1005959.

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In Vitro Characterization of 6S RNA Binding

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Eco Eσ⁷⁰ (100 nM) was incubated with 5′- or 3′-radioactively-labeled 6S RNA (5 nM) in 10 μL of binding buffer (10 mM Tris-HCl, pH 7.0, 100 mM KCl, 10 mM MgCl₂) at 37 °C for 10 min. Subsequently, 1 μL of RNase A (1 ng) or RNase V1 (0.001 U) (Ambion) was added and incubation was continued for 5 min. For the ladder (6S RNA cleaved at G-residues) 0.1 U of RNase T1 was added to the denatured RNA (according to Ambion protocol). Cleavage reactions were stopped by the addition of 20 μL of inactivation buffer (Ambion), and the transcripts were precipitated, suspended in 5 μL of a formamide-containing sample buffer and analyzed on an 8% polyacrylamide/8M urea sequencing gel run in 1× TBE.

Chen J., Wassarman K.M., Feng S., Leon K., Feklistov A., Winkelman J.T., Li Z., Walz T., Campbell E.A, & Darst S.A. (2017). 6S RNA mimics B-form DNA to regulate Escherichia coli RNA polymerase. Molecular cell, 68(2), 388-397.e6.

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Quantitative RNA Analysis by RPA

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Ribonuclease protection assays were performed by using the Ribonuclease Protection Assay (RPA) III kit (Ambion). Labeled riboprobes were gel-purified by 5% PAGE containing 8 M urea in 1XTBE buffer per kit instructions, and hybridized with 10-20 !g total RNA from either barley, yeast, or mouse for 16-18 h at 42 °C. Reaction mixtures were digested with RNase A/T1 (1:100) for 30 min at 37 °C, then stopped with inactivation buffer (Ambion) and protected fragments were precipitated by using 10 !g yeast RNA as a carrier. The protected fragments were separated by 12.5% PAGE containing 8 M urea in 1X TBE buffer. γ-32 ATP (Perkin Elmer Health Sciences) endlabeled Decade Marker (Ambion), prepared per manufacturer's protocol, served as the size standard. Autoradiograms of RPA gels were uniformly scanned at 600 dpi grayscale in a lossless format. The intensity of bands in the 21-24nt range were analyzed using ImageJ (Version 1.49E).

Nething D.B., Mishler‐Elmore J.W., & Held M. (2020). Post-transcriptional regulation of cellulose synthase genes by small RNAs derived from CESA antisense transcripts.

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