Structured Review

HARTMANN ANALYTIC α 32 p atp
AP hydrolyzes [γ- 32 <t>P]ATP</t> 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.
α 32 P Atp, supplied by HARTMANN ANALYTIC, used in various techniques. Bioz Stars score: 93/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Images

1) Product Images from "Binding of ATP to vascular endothelial growth factor isoform VEGF-A165 is essential for inducing proliferation of human umbilical vein endothelial cells"

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

Journal: BMC Biochemistry

doi: 10.1186/1471-2091-12-28

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.
Figure Legend 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.

Techniques Used: 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.
Figure Legend 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.

Techniques Used: 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.
Figure Legend 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.

Techniques Used: Labeling, Incubation, SDS Page, Autoradiography

2) Product Images from "Binding of ATP to vascular endothelial growth factor isoform VEGF-A165 is essential for inducing proliferation of human umbilical vein endothelial cells"

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

Journal: BMC Biochemistry

doi: 10.1186/1471-2091-12-28

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.
Figure Legend 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.

Techniques Used: 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.
Figure Legend 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.

Techniques Used: 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.
Figure Legend 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.

Techniques Used: Labeling, Incubation, SDS Page, Autoradiography

3) Product Images from "Intrinsic regulation of FIC-domain AMP-transferases by oligomerization and automodification"

Article Title: Intrinsic regulation of FIC-domain AMP-transferases by oligomerization and automodification

Journal: Proceedings of the National Academy of Sciences of the United States of America

doi: 10.1073/pnas.1516930113

NmFic E186G adenylylates GyrB. Autoradiographs obtained after incubation of NmFic wt ( Left ) and inhibition-relieved variant NmFic E186G ) ( Right ) with 40 nM α-[ 32 P]ATP, 15 mM MgCl 2 , and E. coli cell lysates overexpressing potential targets as indicated. Incubation was for 1 h at 30 °C.
Figure Legend Snippet: NmFic E186G adenylylates GyrB. Autoradiographs obtained after incubation of NmFic wt ( Left ) and inhibition-relieved variant NmFic E186G ) ( Right ) with 40 nM α-[ 32 P]ATP, 15 mM MgCl 2 , and E. coli cell lysates overexpressing potential targets as indicated. Incubation was for 1 h at 30 °C.

Techniques Used: Incubation, Inhibition, Variant Assay

Related Articles

Synthesized:

Article Title: Dual expression of CCA-adding enzyme and RNase T in Escherichia coli generates a distinct cca growth phenotype with diverse applications
Article Snippet: .. Preparation of in vitro tRNA substrates Radioactively labelled yeast tRNAPhe(GAA) substrate was synthesized via in vitro transcription in the presence of α-32 P ATP (Hartmann Analytic) and purified as described ( , ). .. in vitro nucleotide incorporation assay For qualitative analysis of crude extract activity, 2 ml of an overnight culture were pelleted.

Incubation:

Article Title: Interleukin-6 receptor specific RNA aptamers for cargo delivery into target cells
Article Snippet: Nucleic acids were radioactively labeled during T7 transcription by incorporation of [α-32 P]-ATP (3,000 Ci/mmol, Hartmann Analytic, SCP-207). .. After gel purification, constant amounts of labeled RNA ( < 1 nM) were incubated with increasing amounts of the target protein (0–500 nM) in 1× selection buffer.

Article Title: Binding of ATP to vascular endothelial growth factor isoform VEGF-A165 is essential for inducing proliferation of human umbilical vein endothelial cells
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 ). .. Incubation was performed in 25 mM Tris-HCl (pH 7.5, total volume 15 μL, 37°C, 15 min).

Article Title: Cyclic nucleotides in archaea: Cyclic di‐AMP in the archaeon Haloferax volcanii and its putative role, et al. Cyclic nucleotides in archaea: Cyclic di‐AMP in the archaeon Haloferax volcanii and its putative role
Article Snippet: As substrate, 100 µmol/L unlabeled ATP (Sigma) (solved in 500 mmol/L Tris/HCl, pH 7.5) mixed with [α‐32 P]ATP (Hartmann Analytic) to a final activity of 200 Ci/mmol (final concentration of [α‐32 P]ATP: 55 nmol/L) was used. .. Assays were incubated at 37°C for 1 hr and stopped by adding an equal volume of 0.5 M EDTA.

Activity Assay:

Article Title: Depupylase Dop Requires Inorganic Phosphate in the Active Site for Catalysis *
Article Snippet: .. [α-32 P]ATP was obtained from Hartmann Analytic (Braunschweig, Germany) at a specific activity of 15 TBq (400 Ci)/mmol. .. Polyethyleneimine TLC plates were provided by VWR International.

Article Title: Intrinsic regulation of FIC-domain AMP-transferases by oligomerization and automodification
Article Snippet: .. Adenylylation activity of NmFic was assessed by incubating NmFic with 5 mM ATP, 10 μCi [α-32 P]-ATP (Hartmann Analytic), 25 mM MgCl2 ). .. NmFicE156R and NmFicE156R,Y183F (10 mg/mL and 8 mg/mL, respectively) crystallized in 500 μL of precipitant solution containing 10 mM Tris (pH 7.8) and 100 mM NaCl using the batch crystallization method at 4 °C.

Article Title: The Lid Domain of Caenorhabditis elegans Hsc70 Influences ATP Turnover, Cofactor Binding and Protein Folding Activity
Article Snippet: .. Single-turnover ATPase activity experiments Single-turnover ATPase assays were based on the separation of [α-32 P]-ADP from [α-32 P]-ATP (Hartmann Analytic, Braunschweig, Germany) by thin layer chromatography . .. 30 µl of a solution containing 20 µM CeHsc70 and 30 µM DNJ-13 or BAG-1 in assay buffer (40 mM HEPES/KOH pH 7.5, 150 mM KCl, 5 mM MgCl2 ) were mixed to a final concentration of 5 µM ATP containing 1.0 µCi [α-32 P]-ATP.

Article Title: Cyclic nucleotides in archaea: Cyclic di‐AMP in the archaeon Haloferax volcanii and its putative role, et al. Cyclic nucleotides in archaea: Cyclic di‐AMP in the archaeon Haloferax volcanii and its putative role
Article Snippet: .. As substrate, 100 µmol/L unlabeled ATP (Sigma) (solved in 500 mmol/L Tris/HCl, pH 7.5) mixed with [α‐32 P]ATP (Hartmann Analytic) to a final activity of 200 Ci/mmol (final concentration of [α‐32 P]ATP: 55 nmol/L) was used. .. For testing activity with GTP, 100 µmol/L of unlabeled GTP (Sigma) (dissolved in 500 mmol/L Tris/HCl, pH 7.5) mixed with [α‐32 P]GTP (Hartmann Analytic) to a final activity of 167 Ci/mmol (final concentration of [α‐32 P]GTP: 66 nmol/L) was used.

Expressing:

Article Title: Intrinsic regulation of FIC-domain AMP-transferases by oligomerization and automodification
Article Snippet: Adenylylation assays were performed using cleared cell lysate of ectopically expressing E. coli as described previously ( ) [except using BL21 instead of BL21 (λDE3) cells for overexpression] or using purified proteins as described by Goepfert et al. ( ). .. Adenylylation activity of NmFic was assessed by incubating NmFic with 5 mM ATP, 10 μCi [α-32 P]-ATP (Hartmann Analytic), 25 mM MgCl2 ).

Over Expression:

Article Title: Intrinsic regulation of FIC-domain AMP-transferases by oligomerization and automodification
Article Snippet: Adenylylation assays were performed using cleared cell lysate of ectopically expressing E. coli as described previously ( ) [except using BL21 instead of BL21 (λDE3) cells for overexpression] or using purified proteins as described by Goepfert et al. ( ). .. Adenylylation activity of NmFic was assessed by incubating NmFic with 5 mM ATP, 10 μCi [α-32 P]-ATP (Hartmann Analytic), 25 mM MgCl2 ).

Hybridization:

Article Title: RNase E cleavage shapes the transcriptome of Rhodobacter sphaeroides and strongly impacts phototrophic growth
Article Snippet: .. Oligodeoxynucleotides used for hybridization are listed in Table S1. α-32 [P]-ATP (SRP-301; Hartmann Analytic) and T4 polynucleotide kinase (#EK0031; Fermentas) were used for the end-labeling reaction. .. For detection of SorX, a PCR product (primers listed in Table S1) was labeled with α-[32 P]-dCTP (SRP-205; Hartmann Analytic) using the Prime-a-Gene Labeling System (U1100; Promega) as described in the manufacturer’s manual.

Northern Blot:

Article Title: RNase E cleavage shapes the transcriptome of Rhodobacter sphaeroides and strongly impacts phototrophic growth
Article Snippet: Paragraph title: RNA isolation and Northern blot analysis ... Oligodeoxynucleotides used for hybridization are listed in Table S1. α-32 [P]-ATP (SRP-301; Hartmann Analytic) and T4 polynucleotide kinase (#EK0031; Fermentas) were used for the end-labeling reaction.

Generated:

Article Title: Comparative analysis of Cas6b processing and CRISPR RNA stability
Article Snippet: .. Radiolabeled spacer-repeat-spacer substrates were generated by in vitro run-off transcription reducing the amount of ATP to 2 mM and adding 25 µCi (α-32 P) adenosine triphosphate (ATP) (5,000 Ci/mmol, Hartman Analytic). .. For the production of crRNA substrates, unlabeled spacer-repeat-spacer RNAs were processed with 15 µM Cas6b and purified by phenol-chloroform extraction.

Protein Concentration:

Article Title: Cyclic nucleotides in archaea: Cyclic di‐AMP in the archaeon Haloferax volcanii and its putative role, et al. Cyclic nucleotides in archaea: Cyclic di‐AMP in the archaeon Haloferax volcanii and its putative role
Article Snippet: Final protein concentration of purified DacZ within the assay mixture was 10 µmol/L. .. As substrate, 100 µmol/L unlabeled ATP (Sigma) (solved in 500 mmol/L Tris/HCl, pH 7.5) mixed with [α‐32 P]ATP (Hartmann Analytic) to a final activity of 200 Ci/mmol (final concentration of [α‐32 P]ATP: 55 nmol/L) was used.

Polymerase Chain Reaction:

Article Title: RNase E cleavage shapes the transcriptome of Rhodobacter sphaeroides and strongly impacts phototrophic growth
Article Snippet: Absence of DNA contamination was confirmed by PCR against the gloB gene (RSP_0799). .. Oligodeoxynucleotides used for hybridization are listed in Table S1. α-32 [P]-ATP (SRP-301; Hartmann Analytic) and T4 polynucleotide kinase (#EK0031; Fermentas) were used for the end-labeling reaction.

Article Title: A Temporal Order in 5′- and 3′- Processing of Eukaryotic tRNAHis
Article Snippet: Preparation of tRNA Substrates DNA templates for T7-based in vitro transcription were prepared by overlap extension PCR. .. For internally labeled tRNAHis , transcription was performed in the presence of α-32 P-ATP (Hartman Analytics).

Article Title: Comparative analysis of Cas6b processing and CRISPR RNA stability
Article Snippet: Transcriptions for unlabeled RNAs were performed for 1 h at 37°C in a final volume of 20 µl [40 mM HEPES-KOH (pH 8.0); 22 mM MgCl2 ; 5 mM DTT; 1 mM spermidine; 4 mM UTP, CTP, GTP, ATP; 20 U RNase Inhibitor, 1 µg T7 RNAP, 1 µg PCR product]. .. Radiolabeled spacer-repeat-spacer substrates were generated by in vitro run-off transcription reducing the amount of ATP to 2 mM and adding 25 µCi (α-32 P) adenosine triphosphate (ATP) (5,000 Ci/mmol, Hartman Analytic).

Binding Assay:

Article Title: Interleukin-6 receptor specific RNA aptamers for cargo delivery into target cells
Article Snippet: Filter retention assay (FRA) To investigate the binding of RNA molecules to the target protein, filter retention assays were performed. .. Nucleic acids were radioactively labeled during T7 transcription by incorporation of [α-32 P]-ATP (3,000 Ci/mmol, Hartmann Analytic, SCP-207).

Molecular Weight:

Article Title: Type III restriction endonuclease EcoP15I is a heterotrimeric complex containing one Res subunit with several DNA-binding regions and ATPase activity
Article Snippet: Phusion Hot Start polymerase was purchased from Finnzyme, Prestained Protein Molecular Weight Marker from Fermentas and sinefungin from Sigma-Aldrich. .. [γ-32 P] ATP and [α-32 P] ATP were purchased from Hartmann Analytic GmbH.

Ion Exchange Chromatography:

Article Title: Depupylase Dop Requires Inorganic Phosphate in the Active Site for Catalysis *
Article Snippet: [α-32 P]ATP was obtained from Hartmann Analytic (Braunschweig, Germany) at a specific activity of 15 TBq (400 Ci)/mmol. .. [α-32 P]ATP was obtained from Hartmann Analytic (Braunschweig, Germany) at a specific activity of 15 TBq (400 Ci)/mmol.

Isolation:

Article Title: RNase E cleavage shapes the transcriptome of Rhodobacter sphaeroides and strongly impacts phototrophic growth
Article Snippet: Paragraph title: RNA isolation and Northern blot analysis ... Oligodeoxynucleotides used for hybridization are listed in Table S1. α-32 [P]-ATP (SRP-301; Hartmann Analytic) and T4 polynucleotide kinase (#EK0031; Fermentas) were used for the end-labeling reaction.

Article Title: Comparative analysis of Cas6b processing and CRISPR RNA stability
Article Snippet: PCR reactions with genomic DNA, isolated from Methanococcus maripaludis C5 and containing forward primers encoding the T7 RNA polymerase (RNAP) promoter, yielded spacer(n)-repeat-spacer(n+1) templates for in vitro run-off transcription ( ). .. Radiolabeled spacer-repeat-spacer substrates were generated by in vitro run-off transcription reducing the amount of ATP to 2 mM and adding 25 µCi (α-32 P) adenosine triphosphate (ATP) (5,000 Ci/mmol, Hartman Analytic).

Size-exclusion Chromatography:

Article Title: Depupylase Dop Requires Inorganic Phosphate in the Active Site for Catalysis *
Article Snippet: [α-32 P]ATP was obtained from Hartmann Analytic (Braunschweig, Germany) at a specific activity of 15 TBq (400 Ci)/mmol. .. [α-32 P]ATP was obtained from Hartmann Analytic (Braunschweig, Germany) at a specific activity of 15 TBq (400 Ci)/mmol.

Labeling:

Article Title: Interleukin-6 receptor specific RNA aptamers for cargo delivery into target cells
Article Snippet: .. Nucleic acids were radioactively labeled during T7 transcription by incorporation of [α-32 P]-ATP (3,000 Ci/mmol, Hartmann Analytic, SCP-207). .. After gel purification, constant amounts of labeled RNA ( < 1 nM) were incubated with increasing amounts of the target protein (0–500 nM) in 1× selection buffer.

Article Title: RNase E cleavage shapes the transcriptome of Rhodobacter sphaeroides and strongly impacts phototrophic growth
Article Snippet: Oligodeoxynucleotides used for hybridization are listed in Table S1. α-32 [P]-ATP (SRP-301; Hartmann Analytic) and T4 polynucleotide kinase (#EK0031; Fermentas) were used for the end-labeling reaction. .. For detection of SorX, a PCR product (primers listed in Table S1) was labeled with α-[32 P]-dCTP (SRP-205; Hartmann Analytic) using the Prime-a-Gene Labeling System (U1100; Promega) as described in the manufacturer’s manual.

Article Title: A Temporal Order in 5′- and 3′- Processing of Eukaryotic tRNAHis
Article Snippet: .. For internally labeled tRNAHis , transcription was performed in the presence of α-32 P-ATP (Hartman Analytics). .. In vitro transcripts for Thg1-catalyzed G-1 addition were 5′-labeled using γ-32 P-ATP (Perkin Elmer) and T4 polynucleotide kinase according to the manufacturer’s instructions (NEB).

Article Title: Binding of ATP to vascular endothelial growth factor isoform VEGF-A165 is essential for inducing proliferation of human umbilical vein endothelial cells
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 ). .. Incubation was performed in 25 mM Tris-HCl (pH 7.5, total volume 15 μL, 37°C, 15 min).

Article Title: Comparative analysis of Cas6b processing and CRISPR RNA stability
Article Snippet: Radiolabeled spacer-repeat-spacer substrates were generated by in vitro run-off transcription reducing the amount of ATP to 2 mM and adding 25 µCi (α-32 P) adenosine triphosphate (ATP) (5,000 Ci/mmol, Hartman Analytic). .. The obtained crRNAs were 5′-end labeled using (γ-32 P) ATP (5,000 Ci/mmol) and T4 polynucleotide kinase (PNK) in a volume of 20 µl [15 µl of purified RNA; 2 µl PNK buffer (New England Biolabs) and 25 U T4 PNK (Ambion)] for 1 h at 37°C.

Purification:

Article Title: Dual expression of CCA-adding enzyme and RNase T in Escherichia coli generates a distinct cca growth phenotype with diverse applications
Article Snippet: .. Preparation of in vitro tRNA substrates Radioactively labelled yeast tRNAPhe(GAA) substrate was synthesized via in vitro transcription in the presence of α-32 P ATP (Hartmann Analytic) and purified as described ( , ). .. in vitro nucleotide incorporation assay For qualitative analysis of crude extract activity, 2 ml of an overnight culture were pelleted.

Article Title: Depupylase Dop Requires Inorganic Phosphate in the Active Site for Catalysis *
Article Snippet: [α-32 P]ATP was obtained from Hartmann Analytic (Braunschweig, Germany) at a specific activity of 15 TBq (400 Ci)/mmol. .. [α-32 P]ATP was obtained from Hartmann Analytic (Braunschweig, Germany) at a specific activity of 15 TBq (400 Ci)/mmol.

Article Title: Intrinsic regulation of FIC-domain AMP-transferases by oligomerization and automodification
Article Snippet: Adenylylation assays were performed using cleared cell lysate of ectopically expressing E. coli as described previously ( ) [except using BL21 instead of BL21 (λDE3) cells for overexpression] or using purified proteins as described by Goepfert et al. ( ). .. Adenylylation activity of NmFic was assessed by incubating NmFic with 5 mM ATP, 10 μCi [α-32 P]-ATP (Hartmann Analytic), 25 mM MgCl2 ).

Article Title: Cyclic nucleotides in archaea: Cyclic di‐AMP in the archaeon Haloferax volcanii and its putative role, et al. Cyclic nucleotides in archaea: Cyclic di‐AMP in the archaeon Haloferax volcanii and its putative role
Article Snippet: Final protein concentration of purified DacZ within the assay mixture was 10 µmol/L. .. As substrate, 100 µmol/L unlabeled ATP (Sigma) (solved in 500 mmol/L Tris/HCl, pH 7.5) mixed with [α‐32 P]ATP (Hartmann Analytic) to a final activity of 200 Ci/mmol (final concentration of [α‐32 P]ATP: 55 nmol/L) was used.

Article Title: Comparative analysis of Cas6b processing and CRISPR RNA stability
Article Snippet: Radiolabeled spacer-repeat-spacer substrates were generated by in vitro run-off transcription reducing the amount of ATP to 2 mM and adding 25 µCi (α-32 P) adenosine triphosphate (ATP) (5,000 Ci/mmol, Hartman Analytic). .. For the production of crRNA substrates, unlabeled spacer-repeat-spacer RNAs were processed with 15 µM Cas6b and purified by phenol-chloroform extraction.

Protein Purification:

Article Title: Cyclic nucleotides in archaea: Cyclic di‐AMP in the archaeon Haloferax volcanii and its putative role, et al. Cyclic nucleotides in archaea: Cyclic di‐AMP in the archaeon Haloferax volcanii and its putative role
Article Snippet: DacZ activity assays were performed in a total volume of 25 µl in H. volcanii protein purification buffer supplemented with 10 mmol/L of MnCl2 (respectively 10 mmol/L of MgCl2 ; CaCl2 ; NiCl2 ; CoCl2 for the cofactor dependence experiment). .. As substrate, 100 µmol/L unlabeled ATP (Sigma) (solved in 500 mmol/L Tris/HCl, pH 7.5) mixed with [α‐32 P]ATP (Hartmann Analytic) to a final activity of 200 Ci/mmol (final concentration of [α‐32 P]ATP: 55 nmol/L) was used.

Marker:

Article Title: Type III restriction endonuclease EcoP15I is a heterotrimeric complex containing one Res subunit with several DNA-binding regions and ATPase activity
Article Snippet: Phusion Hot Start polymerase was purchased from Finnzyme, Prestained Protein Molecular Weight Marker from Fermentas and sinefungin from Sigma-Aldrich. .. [γ-32 P] ATP and [α-32 P] ATP were purchased from Hartmann Analytic GmbH.

Polyacrylamide Gel Electrophoresis:

Article Title: Binding of ATP to vascular endothelial growth factor isoform VEGF-A165 is essential for inducing proliferation of human umbilical vein endothelial cells
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 ). .. Proteins were separated by reducing sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE; 17.5%).

SDS Page:

Article Title: Binding of ATP to vascular endothelial growth factor isoform VEGF-A165 is essential for inducing proliferation of human umbilical vein endothelial cells
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 ). .. Proteins were separated by reducing sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE; 17.5%).

Selection:

Article Title: Interleukin-6 receptor specific RNA aptamers for cargo delivery into target cells
Article Snippet: Nucleic acids were radioactively labeled during T7 transcription by incorporation of [α-32 P]-ATP (3,000 Ci/mmol, Hartmann Analytic, SCP-207). .. After gel purification, constant amounts of labeled RNA ( < 1 nM) were incubated with increasing amounts of the target protein (0–500 nM) in 1× selection buffer.

In Vitro:

Article Title: Dual expression of CCA-adding enzyme and RNase T in Escherichia coli generates a distinct cca growth phenotype with diverse applications
Article Snippet: .. Preparation of in vitro tRNA substrates Radioactively labelled yeast tRNAPhe(GAA) substrate was synthesized via in vitro transcription in the presence of α-32 P ATP (Hartmann Analytic) and purified as described ( , ). .. in vitro nucleotide incorporation assay For qualitative analysis of crude extract activity, 2 ml of an overnight culture were pelleted.

Article Title: Intrinsic regulation of FIC-domain AMP-transferases by oligomerization and automodification
Article Snippet: Paragraph title: In Vitro Adenylylation Assay. ... Adenylylation activity of NmFic was assessed by incubating NmFic with 5 mM ATP, 10 μCi [α-32 P]-ATP (Hartmann Analytic), 25 mM MgCl2 ).

Article Title: A Temporal Order in 5′- and 3′- Processing of Eukaryotic tRNAHis
Article Snippet: Preparation of tRNA Substrates DNA templates for T7-based in vitro transcription were prepared by overlap extension PCR. .. For internally labeled tRNAHis , transcription was performed in the presence of α-32 P-ATP (Hartman Analytics).

Article Title: Cyclic nucleotides in archaea: Cyclic di‐AMP in the archaeon Haloferax volcanii and its putative role, et al. Cyclic nucleotides in archaea: Cyclic di‐AMP in the archaeon Haloferax volcanii and its putative role
Article Snippet: Paragraph title: In vitro activity determination of DacZ by thin‐layer chromatography using [α‐32 P]ATP ... As substrate, 100 µmol/L unlabeled ATP (Sigma) (solved in 500 mmol/L Tris/HCl, pH 7.5) mixed with [α‐32 P]ATP (Hartmann Analytic) to a final activity of 200 Ci/mmol (final concentration of [α‐32 P]ATP: 55 nmol/L) was used.

Article Title: Comparative analysis of Cas6b processing and CRISPR RNA stability
Article Snippet: .. Radiolabeled spacer-repeat-spacer substrates were generated by in vitro run-off transcription reducing the amount of ATP to 2 mM and adding 25 µCi (α-32 P) adenosine triphosphate (ATP) (5,000 Ci/mmol, Hartman Analytic). .. For the production of crRNA substrates, unlabeled spacer-repeat-spacer RNAs were processed with 15 µM Cas6b and purified by phenol-chloroform extraction.

Concentration Assay:

Article Title: The Lid Domain of Caenorhabditis elegans Hsc70 Influences ATP Turnover, Cofactor Binding and Protein Folding Activity
Article Snippet: Single-turnover ATPase activity experiments Single-turnover ATPase assays were based on the separation of [α-32 P]-ADP from [α-32 P]-ATP (Hartmann Analytic, Braunschweig, Germany) by thin layer chromatography . .. 30 µl of a solution containing 20 µM CeHsc70 and 30 µM DNJ-13 or BAG-1 in assay buffer (40 mM HEPES/KOH pH 7.5, 150 mM KCl, 5 mM MgCl2 ) were mixed to a final concentration of 5 µM ATP containing 1.0 µCi [α-32 P]-ATP.

Article Title: Cyclic nucleotides in archaea: Cyclic di‐AMP in the archaeon Haloferax volcanii and its putative role, et al. Cyclic nucleotides in archaea: Cyclic di‐AMP in the archaeon Haloferax volcanii and its putative role
Article Snippet: .. As substrate, 100 µmol/L unlabeled ATP (Sigma) (solved in 500 mmol/L Tris/HCl, pH 7.5) mixed with [α‐32 P]ATP (Hartmann Analytic) to a final activity of 200 Ci/mmol (final concentration of [α‐32 P]ATP: 55 nmol/L) was used. .. For testing activity with GTP, 100 µmol/L of unlabeled GTP (Sigma) (dissolved in 500 mmol/L Tris/HCl, pH 7.5) mixed with [α‐32 P]GTP (Hartmann Analytic) to a final activity of 167 Ci/mmol (final concentration of [α‐32 P]GTP: 66 nmol/L) was used.

Thin Layer Chromatography:

Article Title: Depupylase Dop Requires Inorganic Phosphate in the Active Site for Catalysis *
Article Snippet: [α-32 P]ATP was obtained from Hartmann Analytic (Braunschweig, Germany) at a specific activity of 15 TBq (400 Ci)/mmol. .. Polyethyleneimine TLC plates were provided by VWR International.

Article Title: The Lid Domain of Caenorhabditis elegans Hsc70 Influences ATP Turnover, Cofactor Binding and Protein Folding Activity
Article Snippet: .. Single-turnover ATPase activity experiments Single-turnover ATPase assays were based on the separation of [α-32 P]-ADP from [α-32 P]-ATP (Hartmann Analytic, Braunschweig, Germany) by thin layer chromatography . .. 30 µl of a solution containing 20 µM CeHsc70 and 30 µM DNJ-13 or BAG-1 in assay buffer (40 mM HEPES/KOH pH 7.5, 150 mM KCl, 5 mM MgCl2 ) were mixed to a final concentration of 5 µM ATP containing 1.0 µCi [α-32 P]-ATP.

Article Title: Cyclic nucleotides in archaea: Cyclic di‐AMP in the archaeon Haloferax volcanii and its putative role, et al. Cyclic nucleotides in archaea: Cyclic di‐AMP in the archaeon Haloferax volcanii and its putative role
Article Snippet: Paragraph title: In vitro activity determination of DacZ by thin‐layer chromatography using [α‐32 P]ATP ... As substrate, 100 µmol/L unlabeled ATP (Sigma) (solved in 500 mmol/L Tris/HCl, pH 7.5) mixed with [α‐32 P]ATP (Hartmann Analytic) to a final activity of 200 Ci/mmol (final concentration of [α‐32 P]ATP: 55 nmol/L) was used.

End Labeling:

Article Title: RNase E cleavage shapes the transcriptome of Rhodobacter sphaeroides and strongly impacts phototrophic growth
Article Snippet: .. Oligodeoxynucleotides used for hybridization are listed in Table S1. α-32 [P]-ATP (SRP-301; Hartmann Analytic) and T4 polynucleotide kinase (#EK0031; Fermentas) were used for the end-labeling reaction. .. For detection of SorX, a PCR product (primers listed in Table S1) was labeled with α-[32 P]-dCTP (SRP-205; Hartmann Analytic) using the Prime-a-Gene Labeling System (U1100; Promega) as described in the manufacturer’s manual.

Gel Purification:

Article Title: Interleukin-6 receptor specific RNA aptamers for cargo delivery into target cells
Article Snippet: Nucleic acids were radioactively labeled during T7 transcription by incorporation of [α-32 P]-ATP (3,000 Ci/mmol, Hartmann Analytic, SCP-207). .. After gel purification, constant amounts of labeled RNA ( < 1 nM) were incubated with increasing amounts of the target protein (0–500 nM) in 1× selection buffer.

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    HARTMANN ANALYTIC α 32 p atp
    AP hydrolyzes [γ- 32 <t>P]ATP</t> 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.
    α 32 P Atp, supplied by HARTMANN ANALYTIC, used in various techniques. Bioz Stars score: 93/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    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

    NmFic E186G adenylylates GyrB. Autoradiographs obtained after incubation of NmFic wt ( Left ) and inhibition-relieved variant NmFic E186G ) ( Right ) with 40 nM α-[ 32 P]ATP, 15 mM MgCl 2 , and E. coli cell lysates overexpressing potential targets as indicated. Incubation was for 1 h at 30 °C.

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Intrinsic regulation of FIC-domain AMP-transferases by oligomerization and automodification

    doi: 10.1073/pnas.1516930113

    Figure Lengend Snippet: NmFic E186G adenylylates GyrB. Autoradiographs obtained after incubation of NmFic wt ( Left ) and inhibition-relieved variant NmFic E186G ) ( Right ) with 40 nM α-[ 32 P]ATP, 15 mM MgCl 2 , and E. coli cell lysates overexpressing potential targets as indicated. Incubation was for 1 h at 30 °C.

    Article Snippet: Adenylylation activity of NmFic was assessed by incubating NmFic with 5 mM ATP, 10 μCi [α-32 P]-ATP (Hartmann Analytic), 25 mM MgCl2 ).

    Techniques: Incubation, Inhibition, Variant Assay