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HARTMANN ANALYTIC α 32 p adenosine triphosphate atp
α 32 P Adenosine Triphosphate Atp, supplied by HARTMANN ANALYTIC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/α 32 p adenosine triphosphate atp/product/HARTMANN ANALYTIC
Average 86 stars, based on 1 article reviews
Price from $9.99 to $1999.99
α 32 p adenosine triphosphate atp - by Bioz Stars, 2020-04
86/100 stars

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Clone Assay:

Article Title: Characterization of CRISPR RNA processing in Clostridium thermocellum and Methanococcus maripaludis
Article Snippet: Generation of RNA substrates The spacer2–repeat–spacer3 and repeat–spacer27–repeat RNA substrates were generated by in vitro run-off transcription using T7 RNA polymerase and internally labelled using [α-32 P] adenosine triphosphate (ATP) (5000 ci/mmol, Hartman Analytic) ( ). .. Templates for in vitro transcription were obtained by cloning of the pre-crRNA sequences with an upstream T7 RNA polymerase promotor sequence into pUC19 vector.

In Vitro:

Article Title: Characterization of CRISPR RNA processing in Clostridium thermocellum and Methanococcus maripaludis
Article Snippet: .. Generation of RNA substrates The spacer2–repeat–spacer3 and repeat–spacer27–repeat RNA substrates were generated by in vitro run-off transcription using T7 RNA polymerase and internally labelled using [α-32 P] adenosine triphosphate (ATP) (5000 ci/mmol, Hartman Analytic) ( ). .. The repeat RNAs and repeat RNAs with a substitution of the first unprocessed nucleotide against a dexoy nucleotide were synthesized by Eurofins MWG Operon.

Synthesized:

Article Title: Characterization of CRISPR RNA processing in Clostridium thermocellum and Methanococcus maripaludis
Article Snippet: Generation of RNA substrates The spacer2–repeat–spacer3 and repeat–spacer27–repeat RNA substrates were generated by in vitro run-off transcription using T7 RNA polymerase and internally labelled using [α-32 P] adenosine triphosphate (ATP) (5000 ci/mmol, Hartman Analytic) ( ). .. The repeat RNAs and repeat RNAs with a substitution of the first unprocessed nucleotide against a dexoy nucleotide were synthesized by Eurofins MWG Operon.

Generated:

Article Title: Characterization of CRISPR RNA processing in Clostridium thermocellum and Methanococcus maripaludis
Article Snippet: .. Generation of RNA substrates The spacer2–repeat–spacer3 and repeat–spacer27–repeat RNA substrates were generated by in vitro run-off transcription using T7 RNA polymerase and internally labelled using [α-32 P] adenosine triphosphate (ATP) (5000 ci/mmol, Hartman Analytic) ( ). .. The repeat RNAs and repeat RNAs with a substitution of the first unprocessed nucleotide against a dexoy nucleotide were synthesized by Eurofins MWG Operon.

Sequencing:

Article Title: Characterization of CRISPR RNA processing in Clostridium thermocellum and Methanococcus maripaludis
Article Snippet: Generation of RNA substrates The spacer2–repeat–spacer3 and repeat–spacer27–repeat RNA substrates were generated by in vitro run-off transcription using T7 RNA polymerase and internally labelled using [α-32 P] adenosine triphosphate (ATP) (5000 ci/mmol, Hartman Analytic) ( ). .. Templates for in vitro transcription were obtained by cloning of the pre-crRNA sequences with an upstream T7 RNA polymerase promotor sequence into pUC19 vector.

Plasmid Preparation:

Article Title: Characterization of CRISPR RNA processing in Clostridium thermocellum and Methanococcus maripaludis
Article Snippet: Generation of RNA substrates The spacer2–repeat–spacer3 and repeat–spacer27–repeat RNA substrates were generated by in vitro run-off transcription using T7 RNA polymerase and internally labelled using [α-32 P] adenosine triphosphate (ATP) (5000 ci/mmol, Hartman Analytic) ( ). .. Templates for in vitro transcription were obtained by cloning of the pre-crRNA sequences with an upstream T7 RNA polymerase promotor sequence into pUC19 vector.

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    HARTMANN ANALYTIC α 32 p atp
    Endoribonucleolytic activity of Sso aIF5A. (a) Lane 1, [α- 32 P] <t>ATP</t> was loaded on the gel. Lane 2, incubation of 5′-PPP-GGA*-″-3′ RNA for 22 min at 65°C. Lanes 3–14, time course of degradation of 5′-PPP-GGA*-″-3′ RNA at 65°C in the presence of N-His-aIF5A purified from E. coli . The blue arrows indicate the full-length RNA and the single A nucleotide. The red arrow indicates a stable degradation product. (b) Lane 1, RNA size marker. Lane 2, incubation of 5´end-labelled 2508sh RNA for 22 min at 65°C. Lanes 3–14, time course of degradation of 5´end-labelled 2508sh RNA at 65°C in the presence of N-His-aIF5A purified from E. coli . The arrow indicates the accumulation of a stable degradation product. Only the relevant part of the autoradiogram is shown.
    α 32 P Atp, supplied by HARTMANN ANALYTIC, used in various techniques. Bioz Stars score: 94/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/α 32 p atp/product/HARTMANN ANALYTIC
    Average 94 stars, based on 8 article reviews
    Price from $9.99 to $1999.99
    α 32 p atp - by Bioz Stars, 2020-04
    94/100 stars
      Buy from Supplier

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    Endoribonucleolytic activity of Sso aIF5A. (a) Lane 1, [α- 32 P] ATP was loaded on the gel. Lane 2, incubation of 5′-PPP-GGA*-″-3′ RNA for 22 min at 65°C. Lanes 3–14, time course of degradation of 5′-PPP-GGA*-″-3′ RNA at 65°C in the presence of N-His-aIF5A purified from E. coli . The blue arrows indicate the full-length RNA and the single A nucleotide. The red arrow indicates a stable degradation product. (b) Lane 1, RNA size marker. Lane 2, incubation of 5´end-labelled 2508sh RNA for 22 min at 65°C. Lanes 3–14, time course of degradation of 5´end-labelled 2508sh RNA at 65°C in the presence of N-His-aIF5A purified from E. coli . The arrow indicates the accumulation of a stable degradation product. Only the relevant part of the autoradiogram is shown.

    Journal: RNA Biology

    Article Title: Indications for a moonlighting function of translation factor aIF5A in the crenarchaeum Sulfolobus solfataricus

    doi: 10.1080/15476286.2019.1582953

    Figure Lengend Snippet: Endoribonucleolytic activity of Sso aIF5A. (a) Lane 1, [α- 32 P] ATP was loaded on the gel. Lane 2, incubation of 5′-PPP-GGA*-″-3′ RNA for 22 min at 65°C. Lanes 3–14, time course of degradation of 5′-PPP-GGA*-″-3′ RNA at 65°C in the presence of N-His-aIF5A purified from E. coli . The blue arrows indicate the full-length RNA and the single A nucleotide. The red arrow indicates a stable degradation product. (b) Lane 1, RNA size marker. Lane 2, incubation of 5´end-labelled 2508sh RNA for 22 min at 65°C. Lanes 3–14, time course of degradation of 5´end-labelled 2508sh RNA at 65°C in the presence of N-His-aIF5A purified from E. coli . The arrow indicates the accumulation of a stable degradation product. Only the relevant part of the autoradiogram is shown.

    Article Snippet: 10 pmol of dephosphorylated 2508sh RNA were radiolabeled at the 5´-end with T4 Polynucleotide Kinase (Thermo Scientific) in the presence of 15 pmol of [α-32 P]ATP (3000 Ci/mmol, Hartmann Analytic GmbH).

    Techniques: Activity Assay, Incubation, Purification, Marker

    AP hydrolyzes [γ- 32 P]ATP and [α- 32 P]ATP bound to VEGF-A 165 . VEGF-A 165 (15 μM) was labeled with radioactive ATP (5 μCi) in Tris-HCl (pH 7.5) at 37°C for 15 min. After labeling (t = 15 min), 300 ng of AP was added (lane 3 and 4). Incubation of all samples was continued for an additional period of 15 min followed by SDS-PAGE and autoradiography.

    Journal: BMC Biochemistry

    Article Title: Binding of ATP to vascular endothelial growth factor isoform VEGF-A165 is essential for inducing proliferation of human umbilical vein endothelial cells

    doi: 10.1186/1471-2091-12-28

    Figure Lengend Snippet: AP hydrolyzes [γ- 32 P]ATP and [α- 32 P]ATP bound to VEGF-A 165 . VEGF-A 165 (15 μM) was labeled with radioactive ATP (5 μCi) in Tris-HCl (pH 7.5) at 37°C for 15 min. After labeling (t = 15 min), 300 ng of AP was added (lane 3 and 4). Incubation of all samples was continued for an additional period of 15 min followed by SDS-PAGE and autoradiography.

    Article Snippet: Labeling of VEGF-A165 with [γ-32 P]ATP and [α-32 P]ATP For labeling, 3 μg VEGF-A165 (unless otherwise noted) was incubated with 5 μCi each of [γ-32 P]ATP or [α-32 P]ATP (Hartmann Analytic, Braunschweig, Germany) and combined with 0.01 mM non-radioactive ATP (optionally containing 0.1 mM MgCl2 ).

    Techniques: Labeling, Incubation, SDS Page, Autoradiography

    Effect of increased ionic strength on labeling of VEGF-A 165 with [γ- 32 P]ATP and [α- 32 P]ATP . VEGF-A 165 (3 μg) was incubated with radioactive ATP (5 μCi) in Tris-HCl (pH 7.5) at 37°C for 30 min. NaCl (100 mM) was added at times (t) indicated.

    Journal: BMC Biochemistry

    Article Title: Binding of ATP to vascular endothelial growth factor isoform VEGF-A165 is essential for inducing proliferation of human umbilical vein endothelial cells

    doi: 10.1186/1471-2091-12-28

    Figure Lengend Snippet: Effect of increased ionic strength on labeling of VEGF-A 165 with [γ- 32 P]ATP and [α- 32 P]ATP . VEGF-A 165 (3 μg) was incubated with radioactive ATP (5 μCi) in Tris-HCl (pH 7.5) at 37°C for 30 min. NaCl (100 mM) was added at times (t) indicated.

    Article Snippet: Labeling of VEGF-A165 with [γ-32 P]ATP and [α-32 P]ATP For labeling, 3 μg VEGF-A165 (unless otherwise noted) was incubated with 5 μCi each of [γ-32 P]ATP or [α-32 P]ATP (Hartmann Analytic, Braunschweig, Germany) and combined with 0.01 mM non-radioactive ATP (optionally containing 0.1 mM MgCl2 ).

    Techniques: Labeling, Incubation

    Labeling of VEGF-A 165 with [γ- 32 P]ATP and [α- 32 P]ATP . VEGF-A 165 (2 μg) was incubated with radioactive ATP (5 μCi) in Tris-HCl (pH 7.5) at 37°C. MgCl 2 (0.1 mM) was added prior to ATP (+). After 15 min of incubation SDS-PAGE and autoradiography were performed.

    Journal: BMC Biochemistry

    Article Title: Binding of ATP to vascular endothelial growth factor isoform VEGF-A165 is essential for inducing proliferation of human umbilical vein endothelial cells

    doi: 10.1186/1471-2091-12-28

    Figure Lengend Snippet: Labeling of VEGF-A 165 with [γ- 32 P]ATP and [α- 32 P]ATP . VEGF-A 165 (2 μg) was incubated with radioactive ATP (5 μCi) in Tris-HCl (pH 7.5) at 37°C. MgCl 2 (0.1 mM) was added prior to ATP (+). After 15 min of incubation SDS-PAGE and autoradiography were performed.

    Article Snippet: Labeling of VEGF-A165 with [γ-32 P]ATP and [α-32 P]ATP For labeling, 3 μg VEGF-A165 (unless otherwise noted) was incubated with 5 μCi each of [γ-32 P]ATP or [α-32 P]ATP (Hartmann Analytic, Braunschweig, Germany) and combined with 0.01 mM non-radioactive ATP (optionally containing 0.1 mM MgCl2 ).

    Techniques: Labeling, Incubation, SDS Page, Autoradiography

    hMTr2 activity and substrate requirements. Methyltransferase activity: In vitro transcribed RNA-GG molecules with the 32 P-labeled cap01 structure ( A ) were incubated with enzymes as indicated in the presence of SAM. Purified product RNA was digested with RNase T2. Digestion products were resolved on 21% polyacrylamide/8 M urea gel and visualized by autoradiography. BAP protein was used as negative control. RNA with 32 P-labeled cap structure created with the TbMTr2 enzyme was used as a reference. Specificity: autoradiography of two-dimensional chromatograms of 5′-phosphate nucleosides on thin layer cellulose plates. [α- 32 P] ATP-labeled in vitro transcribed cap01-RNA-GA was incubated with SAM in the absence, ( B ) or presence ( C ) of the hMTr2 protein. Product RNA was purified, cleaved by nuclease P1 and the resulting nucleotides were analyzed as described ( 44 ). 5′-monophosphate ribonucleosides of G, A, U, C, Am and m 6 A were used as standards. Substrate requirements: In vitro transcribed RNA-GG molecules with 32 P-labeled cap0 ( D ) or capG ( E) structure were incubated with one of the indicated enzymes, added in a given order, in the presence of SAM. After every modification step RNA molecules were purified by phenol/chloroform extraction and ethanol precipitation. Final products were digested and analyzed as described in legend for panel A. Asterisks indicate positions of 32 P-labeled phosphates.

    Journal: Nucleic Acids Research

    Article Title: 2?-O-ribose methylation of cap2 in human: function and evolution in a horizontally mobile family

    doi: 10.1093/nar/gkr038

    Figure Lengend Snippet: hMTr2 activity and substrate requirements. Methyltransferase activity: In vitro transcribed RNA-GG molecules with the 32 P-labeled cap01 structure ( A ) were incubated with enzymes as indicated in the presence of SAM. Purified product RNA was digested with RNase T2. Digestion products were resolved on 21% polyacrylamide/8 M urea gel and visualized by autoradiography. BAP protein was used as negative control. RNA with 32 P-labeled cap structure created with the TbMTr2 enzyme was used as a reference. Specificity: autoradiography of two-dimensional chromatograms of 5′-phosphate nucleosides on thin layer cellulose plates. [α- 32 P] ATP-labeled in vitro transcribed cap01-RNA-GA was incubated with SAM in the absence, ( B ) or presence ( C ) of the hMTr2 protein. Product RNA was purified, cleaved by nuclease P1 and the resulting nucleotides were analyzed as described ( 44 ). 5′-monophosphate ribonucleosides of G, A, U, C, Am and m 6 A were used as standards. Substrate requirements: In vitro transcribed RNA-GG molecules with 32 P-labeled cap0 ( D ) or capG ( E) structure were incubated with one of the indicated enzymes, added in a given order, in the presence of SAM. After every modification step RNA molecules were purified by phenol/chloroform extraction and ethanol precipitation. Final products were digested and analyzed as described in legend for panel A. Asterisks indicate positions of 32 P-labeled phosphates.

    Article Snippet: For generating internally, 32 P-labeled G-capped transcript RNA-GA [Gppp G p* A pGp(TpCp)12 ], transcription was carried out using pTZ19R-RNA-GA template, prepared as described above, with the addition of 10 μCi of [α-32 P] ATP (3000 Ci/mmol; Hartman Analytic GmbH) and the GpppG cap analog (Epicentre) following the manufacturer instructions.

    Techniques: Activity Assay, In Vitro, Labeling, Incubation, Purification, Autoradiography, Negative Control, Modification, Ethanol Precipitation