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Trevigen fpg
SSB, ALS and 8oxoG repair of UVA-irradiated KSCs and TAs (at 10 J/cm 2 ) using comet assay +- <t>Fpg/T4endoV.</t> The results are expressed as the median % of remaining lesions. The results for 8oxoG and TT-dimers were obtained by subtraction of comet assay results with and without Fpg and T4endoV, respectively; the mean of samples from 3 donors +- SD. Statistical analyses were performed for each population (dotted line for TAs and black line for KSCs) between each time and the previous time; * p
Fpg, supplied by Trevigen, used in various techniques. Bioz Stars score: 92/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/fpg/product/Trevigen
Average 92 stars, based on 12 article reviews
Price from $9.99 to $1999.99
fpg - by Bioz Stars, 2020-07
92/100 stars

Images

1) Product Images from "Keratinocyte stem cells are more resistant to UVA radiation than their direct progeny"

Article Title: Keratinocyte stem cells are more resistant to UVA radiation than their direct progeny

Journal: PLoS ONE

doi: 10.1371/journal.pone.0203863

SSB, ALS and 8oxoG repair of UVA-irradiated KSCs and TAs (at 10 J/cm 2 ) using comet assay +- Fpg/T4endoV. The results are expressed as the median % of remaining lesions. The results for 8oxoG and TT-dimers were obtained by subtraction of comet assay results with and without Fpg and T4endoV, respectively; the mean of samples from 3 donors +- SD. Statistical analyses were performed for each population (dotted line for TAs and black line for KSCs) between each time and the previous time; * p
Figure Legend Snippet: SSB, ALS and 8oxoG repair of UVA-irradiated KSCs and TAs (at 10 J/cm 2 ) using comet assay +- Fpg/T4endoV. The results are expressed as the median % of remaining lesions. The results for 8oxoG and TT-dimers were obtained by subtraction of comet assay results with and without Fpg and T4endoV, respectively; the mean of samples from 3 donors +- SD. Statistical analyses were performed for each population (dotted line for TAs and black line for KSCs) between each time and the previous time; * p

Techniques Used: Irradiation, Single Cell Gel Electrophoresis

SSB, ALS, 8oxoG and thymine dimer induction in UVA-irradiated KSCS and TAs (at 10 J/cm 2 ) by comet assay +- Fpg/T4 endoV. The results are expressed as the median % tail intensity. The results for 8oxoG and TT-dimers were obtained by subtraction of the comet assay results with and without Fpg and T4endoV, respectively; the mean of samples from 3 donors +- SD; * p
Figure Legend Snippet: SSB, ALS, 8oxoG and thymine dimer induction in UVA-irradiated KSCS and TAs (at 10 J/cm 2 ) by comet assay +- Fpg/T4 endoV. The results are expressed as the median % tail intensity. The results for 8oxoG and TT-dimers were obtained by subtraction of the comet assay results with and without Fpg and T4endoV, respectively; the mean of samples from 3 donors +- SD; * p

Techniques Used: Irradiation, Single Cell Gel Electrophoresis

2) Product Images from "Guanine-specific DNA damage induced by ?-irradiated histone"

Article Title: Guanine-specific DNA damage induced by ?-irradiated histone

Journal: Biochemical Journal

doi: 10.1042/BJ20050186

Effect of Fpg treatment on DNA damage induced by histone H1-hydroperoxides in the presence of Cu(I) Reaction mixtures contained the 32 P-5′-end-labelled 337 bp fragment, calf thymus DNA (20 μM in DNA bases), the indicated concentrations of histone H1-hydroperoxides and 20 μM Cu(I) in 200 μl of 10 mM sodium phosphate buffer (pH 7.4) containing 5 μM DTPA. The mixture was incubated at 37 °C for 60 min. The DNA fragments were then treated with Fpg and electrophoresed on an 8% polyacrylamide/8 M urea gel (12 cm×16 cm), and autoradiograms were obtained by exposing an X-ray film to the gel.
Figure Legend Snippet: Effect of Fpg treatment on DNA damage induced by histone H1-hydroperoxides in the presence of Cu(I) Reaction mixtures contained the 32 P-5′-end-labelled 337 bp fragment, calf thymus DNA (20 μM in DNA bases), the indicated concentrations of histone H1-hydroperoxides and 20 μM Cu(I) in 200 μl of 10 mM sodium phosphate buffer (pH 7.4) containing 5 μM DTPA. The mixture was incubated at 37 °C for 60 min. The DNA fragments were then treated with Fpg and electrophoresed on an 8% polyacrylamide/8 M urea gel (12 cm×16 cm), and autoradiograms were obtained by exposing an X-ray film to the gel.

Techniques Used: Incubation

Autoradiogram of 32 P-labelled DNA fragments incubated with various histone protein-hydroperoxides in the presence of Cu(I) The reaction mixtures contained the 32 P-5′-end-labelled 337 bp DNA fragment, calf thymus DNA (20 μM in DNA bases), 0.7 μM of histone protein-hydroperoxides (histone H1, H2A, H3 and H4) and 20 μM CuCl in 200 μl of 10 mM sodium phosphate buffer (pH 7.4) containing 5 μM DTPA. After incubation at 37 °C for 60 min, DNA fragments were treated with 10 units of Fpg protein at 37 °C for 120 min. The DNA fragments were electrophoresed on an 8% (w/v) polyacrylamide/8 M urea gel (12 cm×16 cm), and autoradiograms were obtained by exposing an X-ray film to the gel. Oligonucleotides generated by hydroperoxide-induced DNA damage are indicated by arrows. The bands above these oligonucleotides are single- and double-strand intact DNA fragments.
Figure Legend Snippet: Autoradiogram of 32 P-labelled DNA fragments incubated with various histone protein-hydroperoxides in the presence of Cu(I) The reaction mixtures contained the 32 P-5′-end-labelled 337 bp DNA fragment, calf thymus DNA (20 μM in DNA bases), 0.7 μM of histone protein-hydroperoxides (histone H1, H2A, H3 and H4) and 20 μM CuCl in 200 μl of 10 mM sodium phosphate buffer (pH 7.4) containing 5 μM DTPA. After incubation at 37 °C for 60 min, DNA fragments were treated with 10 units of Fpg protein at 37 °C for 120 min. The DNA fragments were electrophoresed on an 8% (w/v) polyacrylamide/8 M urea gel (12 cm×16 cm), and autoradiograms were obtained by exposing an X-ray film to the gel. Oligonucleotides generated by hydroperoxide-induced DNA damage are indicated by arrows. The bands above these oligonucleotides are single- and double-strand intact DNA fragments.

Techniques Used: Incubation, Generated

Site specificity of DNA damage induced by histone H1-hydroperoxides in the presence of Cu(I) The 32 P-5′-end-labelled 261 bp fragment ( A ) and 211 bp fragment ( B ) were exposed to 0.7 μM histone H1-hydroperoxides, 20 μM Cu(I) and calf thymus DNA (20 μM in DNA bases) in 200 μl of 10 mM sodium phosphate buffer (pH 7.4) containing 5 μM DTPA. After incubation at 37 °C for 60 min, DNA fragments were treated with 10 units of Fpg protein at 37 °C for 120 min. The DNA fragments were electrophoresed on an 8% polyacrylamide/8 M urea gel using a DNA-sequencing system and visualized by autoradiography. The relative amounts of oligonucleotide were measured by scanning the autoradiogram with a laser densitometer. Horizontal axis: the nucleotide number of the human c-Ha- ras -1 proto-oncogene ( A ) and p53 tumour suppressor gene ( B ).
Figure Legend Snippet: Site specificity of DNA damage induced by histone H1-hydroperoxides in the presence of Cu(I) The 32 P-5′-end-labelled 261 bp fragment ( A ) and 211 bp fragment ( B ) were exposed to 0.7 μM histone H1-hydroperoxides, 20 μM Cu(I) and calf thymus DNA (20 μM in DNA bases) in 200 μl of 10 mM sodium phosphate buffer (pH 7.4) containing 5 μM DTPA. After incubation at 37 °C for 60 min, DNA fragments were treated with 10 units of Fpg protein at 37 °C for 120 min. The DNA fragments were electrophoresed on an 8% polyacrylamide/8 M urea gel using a DNA-sequencing system and visualized by autoradiography. The relative amounts of oligonucleotide were measured by scanning the autoradiogram with a laser densitometer. Horizontal axis: the nucleotide number of the human c-Ha- ras -1 proto-oncogene ( A ) and p53 tumour suppressor gene ( B ).

Techniques Used: Incubation, DNA Sequencing, Autoradiography

3) Product Images from "Mitochondrial base excision repair assays"

Article Title: Mitochondrial base excision repair assays

Journal: Methods (San Diego, Calif.)

doi: 10.1016/j.ymeth.2010.02.020

Endogenous oxidative damage is overestimated in mtDNA due to oxidative lesions generated from the mtDNA purification process. Fpg/Southern blot analysis was used to measure 8-oxoG levels in nDNA as well as in mtDNA with and without mitochondrial isolation;
Figure Legend Snippet: Endogenous oxidative damage is overestimated in mtDNA due to oxidative lesions generated from the mtDNA purification process. Fpg/Southern blot analysis was used to measure 8-oxoG levels in nDNA as well as in mtDNA with and without mitochondrial isolation;

Techniques Used: Generated, Purification, Southern Blot, Isolation

4) Product Images from "NEIL2 Protects against Oxidative DNA Damage Induced by Sidestream Smoke in Human Cells"

Article Title: NEIL2 Protects against Oxidative DNA Damage Induced by Sidestream Smoke in Human Cells

Journal: PLoS ONE

doi: 10.1371/journal.pone.0090261

Increased oxidative DNA damage in various human cells. Following exposure to a series of dilutions of SSS–laden DMEM for 24 h, DNA isolation and digestion with Fpg and Nei followed by qPCR for amplification of HPRT large and a short fragment (LA-QPCR assay). Three cell lines, HEK293, hPF and BEAS-2B, all exhibited a dose-response relationship between levels of oxidative DNA damage and the SSS doses used in the LA-QPCR assay. PE: puff equivalents (the smoke from one puff in 1 ml of medium).
Figure Legend Snippet: Increased oxidative DNA damage in various human cells. Following exposure to a series of dilutions of SSS–laden DMEM for 24 h, DNA isolation and digestion with Fpg and Nei followed by qPCR for amplification of HPRT large and a short fragment (LA-QPCR assay). Three cell lines, HEK293, hPF and BEAS-2B, all exhibited a dose-response relationship between levels of oxidative DNA damage and the SSS doses used in the LA-QPCR assay. PE: puff equivalents (the smoke from one puff in 1 ml of medium).

Techniques Used: DNA Extraction, Real-time Polymerase Chain Reaction, Amplification

Effect of NEIL2 knockdown on induction of oxidative DNA damage by exposure to SSS for 24 HPRT gene using LA-QPCR. (A) left: NEIL2 knockdown from hPF was tested by qRT-PCR; right: Western blots showing shRNA-mediated knockdown of NEIL2 in HEK293 cells expressing Flag-NEIL2. (B) Cultured hPF with NEIL2 knockdown were treated with 3 doses of SSS extract as compared to the control followed by DNA isolation and digestion with Fpg and Nei as described in Materials and Methods. The DNA was used as template for HPRT large and short fragment amplification by LA-QPCR assay. The histogram depicts the mean % of DNA tail ± SEM of 3 to 4 independent experiments. The symbol * means that the differences between the Control and NEIL 2 knockdown were statistically significant (P
Figure Legend Snippet: Effect of NEIL2 knockdown on induction of oxidative DNA damage by exposure to SSS for 24 HPRT gene using LA-QPCR. (A) left: NEIL2 knockdown from hPF was tested by qRT-PCR; right: Western blots showing shRNA-mediated knockdown of NEIL2 in HEK293 cells expressing Flag-NEIL2. (B) Cultured hPF with NEIL2 knockdown were treated with 3 doses of SSS extract as compared to the control followed by DNA isolation and digestion with Fpg and Nei as described in Materials and Methods. The DNA was used as template for HPRT large and short fragment amplification by LA-QPCR assay. The histogram depicts the mean % of DNA tail ± SEM of 3 to 4 independent experiments. The symbol * means that the differences between the Control and NEIL 2 knockdown were statistically significant (P

Techniques Used: Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Western Blot, shRNA, Expressing, Cell Culture, DNA Extraction, Amplification

Effect of NEIL2 knockdown on induction of oxidative DNA damage by exposure to SSS for 24h in the human POLB gene using LA-QPCR. (A) Cultured primary hPF with NEIL2 knockdown were treated with 3 doses of SSS extract as compared to the control on the left, followed by processing of the DNA with Fpg and Nei. The DNA was used for Polβ ( POLB ) large and short fragment amplification by LA-QPCR. Quantification was with ImageQuant (Molecular Dynamics). The histogram depicts the mean % of DNA tail ± SEM of 3 to 4 independent experiments. The symbol * means that the differences between the Control and NEIL2 knockdown were statistically significant (P
Figure Legend Snippet: Effect of NEIL2 knockdown on induction of oxidative DNA damage by exposure to SSS for 24h in the human POLB gene using LA-QPCR. (A) Cultured primary hPF with NEIL2 knockdown were treated with 3 doses of SSS extract as compared to the control on the left, followed by processing of the DNA with Fpg and Nei. The DNA was used for Polβ ( POLB ) large and short fragment amplification by LA-QPCR. Quantification was with ImageQuant (Molecular Dynamics). The histogram depicts the mean % of DNA tail ± SEM of 3 to 4 independent experiments. The symbol * means that the differences between the Control and NEIL2 knockdown were statistically significant (P

Techniques Used: Real-time Polymerase Chain Reaction, Cell Culture, Amplification

5) Product Images from "Overexpression of PCNA Attenuates Oxidative Stress-Caused Delay of Gap-Filling during Repair of UV-Induced DNA Damage"

Article Title: Overexpression of PCNA Attenuates Oxidative Stress-Caused Delay of Gap-Filling during Repair of UV-Induced DNA Damage

Journal: Journal of Nucleic Acids

doi: 10.1155/2017/8154646

The effect of Fen-1 knockdown. (a) On repair of oxidative DNA damage. Top: western analysis of Fen-1 protein in AGS cells transfected with pLKO-1 sh Fen-1 to knock down Fen-1 or control vector; levels of PCNA protein were used as a loading control. Bottom: comet assay. AGS cells were treated with 20 μ M H 2 O 2 for indicated periods of time before being harvested for comet-Fpg/Endo III assay. (b) On DNA replication. S-phase cells were marked.
Figure Legend Snippet: The effect of Fen-1 knockdown. (a) On repair of oxidative DNA damage. Top: western analysis of Fen-1 protein in AGS cells transfected with pLKO-1 sh Fen-1 to knock down Fen-1 or control vector; levels of PCNA protein were used as a loading control. Bottom: comet assay. AGS cells were treated with 20 μ M H 2 O 2 for indicated periods of time before being harvested for comet-Fpg/Endo III assay. (b) On DNA replication. S-phase cells were marked.

Techniques Used: Western Blot, Transfection, Plasmid Preparation, Single Cell Gel Electrophoresis

The effect of PCNA knockdown on the repair of oxidative DNA damage. (a) Comet assay. AGS cells transfected with pLKO-1 shPCNA to knock down PCNA or control vector were treated with 20 μ M H 2 O 2 for indicated periods of time before being harvested for comet-Fpg/Endo III assay to monitor the levels of H 2 O 2 -induced DNA lesions in cells. Bottom panel: western analysis of PCNA protein with levels of β -actin protein as a loading control. (b) Immunostaining analysis and (c) ELISA. Cells transfected with pLKO-1 shPCNA or control vector were treated with 1 mM H 2 O 2 for 1 and 4 h before being harvested for immunofluorescence and ELISA assays of 8-oxo-dG. Typical images were shown. N for control, that is, cells without treatment. ∗∗ for p
Figure Legend Snippet: The effect of PCNA knockdown on the repair of oxidative DNA damage. (a) Comet assay. AGS cells transfected with pLKO-1 shPCNA to knock down PCNA or control vector were treated with 20 μ M H 2 O 2 for indicated periods of time before being harvested for comet-Fpg/Endo III assay to monitor the levels of H 2 O 2 -induced DNA lesions in cells. Bottom panel: western analysis of PCNA protein with levels of β -actin protein as a loading control. (b) Immunostaining analysis and (c) ELISA. Cells transfected with pLKO-1 shPCNA or control vector were treated with 1 mM H 2 O 2 for 1 and 4 h before being harvested for immunofluorescence and ELISA assays of 8-oxo-dG. Typical images were shown. N for control, that is, cells without treatment. ∗∗ for p

Techniques Used: Single Cell Gel Electrophoresis, Transfection, Plasmid Preparation, Western Blot, Immunostaining, Enzyme-linked Immunosorbent Assay, Immunofluorescence

6) Product Images from "Keratinocyte stem cells are more resistant to UVA radiation than their direct progeny"

Article Title: Keratinocyte stem cells are more resistant to UVA radiation than their direct progeny

Journal: PLoS ONE

doi: 10.1371/journal.pone.0203863

SSB, ALS and 8oxoG repair of UVA-irradiated KSCs and TAs (at 10 J/cm 2 ) using comet assay +- Fpg/T4endoV. The results are expressed as the median % of remaining lesions. The results for 8oxoG and TT-dimers were obtained by subtraction of comet assay results with and without Fpg and T4endoV, respectively; the mean of samples from 3 donors +- SD. Statistical analyses were performed for each population (dotted line for TAs and black line for KSCs) between each time and the previous time; * p
Figure Legend Snippet: SSB, ALS and 8oxoG repair of UVA-irradiated KSCs and TAs (at 10 J/cm 2 ) using comet assay +- Fpg/T4endoV. The results are expressed as the median % of remaining lesions. The results for 8oxoG and TT-dimers were obtained by subtraction of comet assay results with and without Fpg and T4endoV, respectively; the mean of samples from 3 donors +- SD. Statistical analyses were performed for each population (dotted line for TAs and black line for KSCs) between each time and the previous time; * p

Techniques Used: Irradiation, Single Cell Gel Electrophoresis

SSB, ALS, 8oxoG and thymine dimer induction in UVA-irradiated KSCS and TAs (at 10 J/cm 2 ) by comet assay +- Fpg/T4 endoV. The results are expressed as the median % tail intensity. The results for 8oxoG and TT-dimers were obtained by subtraction of the comet assay results with and without Fpg and T4endoV, respectively; the mean of samples from 3 donors +- SD; * p
Figure Legend Snippet: SSB, ALS, 8oxoG and thymine dimer induction in UVA-irradiated KSCS and TAs (at 10 J/cm 2 ) by comet assay +- Fpg/T4 endoV. The results are expressed as the median % tail intensity. The results for 8oxoG and TT-dimers were obtained by subtraction of the comet assay results with and without Fpg and T4endoV, respectively; the mean of samples from 3 donors +- SD; * p

Techniques Used: Irradiation, Single Cell Gel Electrophoresis

Related Articles

Alkaline Single Cell Gel Electrophoresis:

Article Title: Overexpression of PCNA Attenuates Oxidative Stress-Caused Delay of Gap-Filling during Repair of UV-Induced DNA Damage
Article Snippet: .. Comet Assay with Enzymes To measure oxidative DNA adducts with the alkaline comet assay, we performed an additional step immediately after the cell lysis step, that is, incubating the slides containing the nucleoids with Endo III and Fpg (from Trevigen Co.; 2 units of each enzyme per slide in buffer with 10 mM Tris-HCl pH 7.4) as described previously [ ]. ..

Incubation:

Article Title: TRANSGENIC OVEREXPRESSION OF NEUROGLOBIN ATTENUATES FORMATION OF SMOKE INHALATION-INDUCED OXIDATIVE DNA DAMAGE, IN VIVO, IN THE MOUSE BRAIN
Article Snippet: .. Accordingly, duplicate slide sets were used: all slides were equilibrated with the Fpg reaction buffer (10 mM HEPES-KOH, 100 mM KCl, 0.1 mg/ml BSA, pH 7.4) and then incubated either with or without Fpg (cat#4040-100-01, Gaithersburg, Trevigen, MD). .. Subsequently slides were incubated 40 min in alkaline buffer (0.3 M NaOH, 1 mM EDTA), electrophoresed at 1.2 V/cm (300 mA, 25 V), washed 3 times in neutralization buffer (0.4 M Tris, pH 7.5) and stained with 20 µg/ml ethidium bromide.

other:

Article Title: Guanine-specific DNA damage induced by ?-irradiated histone
Article Snippet: Nuclease P1 (400 units/mg) was purchased from Yamasa Shoyu (Chiba, Japan), Fpg ( Escherichia coli formamidopyrimidine-DNA glycosylase) from Trevigen (Gaithersburg, MD, U.S.A.) and histone H1 (subgroup f1-lysine-rich fraction, calf thymus) and other histone proteins from Sigma (St. Louis, MO, U.S.A.).

Single Cell Gel Electrophoresis:

Article Title: Overexpression of PCNA Attenuates Oxidative Stress-Caused Delay of Gap-Filling during Repair of UV-Induced DNA Damage
Article Snippet: .. Comet Assay with Enzymes To measure oxidative DNA adducts with the alkaline comet assay, we performed an additional step immediately after the cell lysis step, that is, incubating the slides containing the nucleoids with Endo III and Fpg (from Trevigen Co.; 2 units of each enzyme per slide in buffer with 10 mM Tris-HCl pH 7.4) as described previously [ ]. ..

Lysis:

Article Title: Keratinocyte stem cells are more resistant to UVA radiation than their direct progeny
Article Snippet: .. For revealing oxidized purines and pyrimidine dimers, Fpg (Trevigen, Gaithersburg, MD, USA) and T4endoV (BioLabs, Ipswich, MA, USA) were added after the lysis step at 0.05 U/μL for 45 min at 37°C. .. The slides were stained with GelRed, and comet analysis was performed using the image analysis Komet 6.0 software (Kinetic Imaging Ltd., Andor Technology plc., Belfast, Ireland).

Article Title: Overexpression of PCNA Attenuates Oxidative Stress-Caused Delay of Gap-Filling during Repair of UV-Induced DNA Damage
Article Snippet: .. Comet Assay with Enzymes To measure oxidative DNA adducts with the alkaline comet assay, we performed an additional step immediately after the cell lysis step, that is, incubating the slides containing the nucleoids with Endo III and Fpg (from Trevigen Co.; 2 units of each enzyme per slide in buffer with 10 mM Tris-HCl pH 7.4) as described previously [ ]. ..

Activity Assay:

Article Title: NEIL2 Protects against Oxidative DNA Damage Induced by Sidestream Smoke in Human Cells
Article Snippet: .. After quantification equal amounts of genomic DNA were digested with two E. coli BER enzymes, Fpg (Trevigen, Gaithersburg, MD) and Nei (provided from Dr. Tapas lab), which are able to remove a variety of oxidized purine and pyrimidine oxidized DNA bases and induce strand breaks by cleaving the phosphodiester bond with their associated AP lyase activity. ..

Chloramphenicol Acetyltransferase Assay:

Article Title: TRANSGENIC OVEREXPRESSION OF NEUROGLOBIN ATTENUATES FORMATION OF SMOKE INHALATION-INDUCED OXIDATIVE DNA DAMAGE, IN VIVO, IN THE MOUSE BRAIN
Article Snippet: .. Accordingly, duplicate slide sets were used: all slides were equilibrated with the Fpg reaction buffer (10 mM HEPES-KOH, 100 mM KCl, 0.1 mg/ml BSA, pH 7.4) and then incubated either with or without Fpg (cat4040-100-01, Gaithersburg, Trevigen, MD). .. Subsequently slides were incubated 40 min in alkaline buffer (0.3 M NaOH, 1 mM EDTA), electrophoresed at 1.2 V/cm (300 mA, 25 V), washed 3 times in neutralization buffer (0.4 M Tris, pH 7.5) and stained with 20 µg/ml ethidium bromide.

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    Trevigen fpg enzyme
    Correlation between the levels of <t>DNA</t> damage obtained in different assays. Correlation of the values obtained in standard comet assay and (A) buffer exposed slides, (B) <t>Fpg</t> exposed slides and (C) net 8-oxoG sites; full line—regression line; dashed line—95 confidence level.
    Fpg Enzyme, supplied by Trevigen, used in various techniques. Bioz Stars score: 89/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/fpg enzyme/product/Trevigen
    Average 89 stars, based on 5 article reviews
    Price from $9.99 to $1999.99
    fpg enzyme - by Bioz Stars, 2020-07
    89/100 stars
      Buy from Supplier

    85
    Trevigen e coli fpg protein
    <t>DNA</t> lesions detected as FPGSS in the c- fos gene. DNA base lesions were detected in genomic DNA from 24 mouse brains that underwent 90 min of ischemia and various durations (min, except 2 h) of reperfusion as indicated on top of each lane. Each lane contains DNA from a pool of four animals having the same reperfusion time. The <t>Fpg</t> protein-treated DNA samples (+) were resolved in parallel with the same DNA samples without Fpg protein treatment (-). The blot was hybridized to a cRNA probe (for the nontranscribed strand) from the c- fos ). The blot was then boiled to remove 32 ), and are not shown.
    E Coli Fpg Protein, supplied by Trevigen, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/e coli fpg protein/product/Trevigen
    Average 85 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    e coli fpg protein - by Bioz Stars, 2020-07
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    85
    Trevigen fpg glycosylase
    NF-κB binds specifically at the nucleosomal ends, but not at the nucleosomal dyad. ( A ) Upper panel : EMSA of NF-κB binding to naked 601_D 7 DNA (left) or to 601_D 7 nucleosome (right). Naked 32 P-end labeled 601_D 7 DNA or nucleosomes were incubated with increasing amount of NF-κB (1.5 fold serial dilution of NF-κB was performed starting with saturating amounts of 100 nM for naked DNA and 400 nM for nucleosome). The aliquots of the reaction mixtures were run on a 5% native PAGE. The positions of free DNA and nucleosomes are indicated, “cplx.” represents NF-κB – DNA/nuc complexes; lower panel: UV laser footprinting patterns of the NF-κB-DNA and NF-κB-nucleosomes complexes. The respective remaining mixtures were irradiated with a single 5 nanoseconds UV laser 266 nm pulse ( E pulse ∼0.1 J/cm 2 ), DNA was purified from the samples and then treated with <t>Fpg</t> <t>glycosylase.</t> The cleaved DNA fragments were separated on 8% sequencing gel and visualized by autoradiography; (*), NF-κB footprint at the high-affinity NF-κB binding site; (♦), NF-κB footprints at low-affinity sites. A schematic presentation of the nucleosomes is shown on the right side; the double headed arrow indicates the nucleosomal dyad. ( B ) UV laser footprinting of NF-κB bound to either naked 601_ D 7 -DNA (lanes 1 and 2) or to 601_D 7 -nucleosome (lanes 3 and 4) repeated with saturated amounts of NF-κB as indicated. ( C ) Same as (B), but for naked 601_D 0 DNA and 601_D 0 -nucleosomes. Note the absence of specific NF-κB footprint at the dyad in the case of the nucleosome.
    Fpg Glycosylase, supplied by Trevigen, used in various techniques. Bioz Stars score: 85/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/fpg glycosylase/product/Trevigen
    Average 85 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    fpg glycosylase - by Bioz Stars, 2020-07
    85/100 stars
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    Image Search Results


    Correlation between the levels of DNA damage obtained in different assays. Correlation of the values obtained in standard comet assay and (A) buffer exposed slides, (B) Fpg exposed slides and (C) net 8-oxoG sites; full line—regression line; dashed line—95 confidence level.

    Journal: PLoS ONE

    Article Title: Evaluation of Genotoxic Pressure along the Sava River

    doi: 10.1371/journal.pone.0162450

    Figure Lengend Snippet: Correlation between the levels of DNA damage obtained in different assays. Correlation of the values obtained in standard comet assay and (A) buffer exposed slides, (B) Fpg exposed slides and (C) net 8-oxoG sites; full line—regression line; dashed line—95 confidence level.

    Article Snippet: In all cases digestion with Fpg enzyme led to significant increase of DNA damage in comparison with corresponding standard alkaline and buffer treated control.

    Techniques: Single Cell Gel Electrophoresis

    DNA lesions detected as FPGSS in the c- fos gene. DNA base lesions were detected in genomic DNA from 24 mouse brains that underwent 90 min of ischemia and various durations (min, except 2 h) of reperfusion as indicated on top of each lane. Each lane contains DNA from a pool of four animals having the same reperfusion time. The Fpg protein-treated DNA samples (+) were resolved in parallel with the same DNA samples without Fpg protein treatment (-). The blot was hybridized to a cRNA probe (for the nontranscribed strand) from the c- fos ). The blot was then boiled to remove 32 ), and are not shown.

    Journal: Journal of neurochemistry

    Article Title: Up-Regulation of Base Excision Repair Activity for 8-Hydroxy-2?-Deoxyguanosine in the Mouse Brain After Forebrain Ischemia--Reperfusion

    doi:

    Figure Lengend Snippet: DNA lesions detected as FPGSS in the c- fos gene. DNA base lesions were detected in genomic DNA from 24 mouse brains that underwent 90 min of ischemia and various durations (min, except 2 h) of reperfusion as indicated on top of each lane. Each lane contains DNA from a pool of four animals having the same reperfusion time. The Fpg protein-treated DNA samples (+) were resolved in parallel with the same DNA samples without Fpg protein treatment (-). The blot was hybridized to a cRNA probe (for the nontranscribed strand) from the c- fos ). The blot was then boiled to remove 32 ), and are not shown.

    Article Snippet: DNA from four animals at each reperfusion time point was pooled, and a total of 24 μ g of DNA from each time point was tested for the presence of DNA base lesions sensitive to digestion by E. coli Fpg protein, which removes oh8dG lesions, as well as 5-hydroxycytosine and 2,6-diamino-4-hydroxy-5-formamidopyrimidine (open ring of dG modification lesions).

    Techniques:

    Activation of endogenous OGG1 activities in the nuclear extracts of mouse brain after FbIR. The endogenous nuclear OGG1 activity was measured by incubating 100 fmol of 32 P-labeled oligoZ (oh8dG/dC) duplex with 30 μ g of nuclear protein from the control (sham-operated without FbIR) and FbIR animals. The oligoZ duplex incubated with Fpg protein (lane marked “Fpg,” 1 U/assay) or without enzymes (lane labeled as “O”) was also included in each experiment. Results from control animals (n = 6) and FbIR animals (n = 3 each for 90/30, 90/60, 90/120, and 90/240) are shown in A (reaction in the sample from one animal per lane). Results from a second experiment in which pooled nuclear extracts from two animals were used in each reaction and then resolved in each lane are presented in B . A total of 46 animals were used in B: 12 animals in the control group, 6 animals each with FbIR of 90/0, 90/10, or 90/20, 8 animals with FbIR of 90/30, and 4 animals each with FbIR of 90/60 or 90/120. Arrowheads indicate the positions of the major 11-mer digested products of the mouse nuclear extracts; arrows indicate the 11-mer with 3′-PO 4 end (product of Fpg protein). The results in B (excluding the data at 120 min due to the variation in A) were quantitatively analyzed and presented in C . Asterisks indicate statistically significant ( p

    Journal: Journal of neurochemistry

    Article Title: Up-Regulation of Base Excision Repair Activity for 8-Hydroxy-2?-Deoxyguanosine in the Mouse Brain After Forebrain Ischemia--Reperfusion

    doi:

    Figure Lengend Snippet: Activation of endogenous OGG1 activities in the nuclear extracts of mouse brain after FbIR. The endogenous nuclear OGG1 activity was measured by incubating 100 fmol of 32 P-labeled oligoZ (oh8dG/dC) duplex with 30 μ g of nuclear protein from the control (sham-operated without FbIR) and FbIR animals. The oligoZ duplex incubated with Fpg protein (lane marked “Fpg,” 1 U/assay) or without enzymes (lane labeled as “O”) was also included in each experiment. Results from control animals (n = 6) and FbIR animals (n = 3 each for 90/30, 90/60, 90/120, and 90/240) are shown in A (reaction in the sample from one animal per lane). Results from a second experiment in which pooled nuclear extracts from two animals were used in each reaction and then resolved in each lane are presented in B . A total of 46 animals were used in B: 12 animals in the control group, 6 animals each with FbIR of 90/0, 90/10, or 90/20, 8 animals with FbIR of 90/30, and 4 animals each with FbIR of 90/60 or 90/120. Arrowheads indicate the positions of the major 11-mer digested products of the mouse nuclear extracts; arrows indicate the 11-mer with 3′-PO 4 end (product of Fpg protein). The results in B (excluding the data at 120 min due to the variation in A) were quantitatively analyzed and presented in C . Asterisks indicate statistically significant ( p

    Article Snippet: Although the OGG1/MMH protein is the mammalian equivalent of E. coli Fpg protein, it is different from Fpg protein in terms of structure and substrate specificity.

    Techniques: Activation Assay, Activity Assay, Labeling, Incubation

    NF-κB binds specifically at the nucleosomal ends, but not at the nucleosomal dyad. ( A ) Upper panel : EMSA of NF-κB binding to naked 601_D 7 DNA (left) or to 601_D 7 nucleosome (right). Naked 32 P-end labeled 601_D 7 DNA or nucleosomes were incubated with increasing amount of NF-κB (1.5 fold serial dilution of NF-κB was performed starting with saturating amounts of 100 nM for naked DNA and 400 nM for nucleosome). The aliquots of the reaction mixtures were run on a 5% native PAGE. The positions of free DNA and nucleosomes are indicated, “cplx.” represents NF-κB – DNA/nuc complexes; lower panel: UV laser footprinting patterns of the NF-κB-DNA and NF-κB-nucleosomes complexes. The respective remaining mixtures were irradiated with a single 5 nanoseconds UV laser 266 nm pulse ( E pulse ∼0.1 J/cm 2 ), DNA was purified from the samples and then treated with Fpg glycosylase. The cleaved DNA fragments were separated on 8% sequencing gel and visualized by autoradiography; (*), NF-κB footprint at the high-affinity NF-κB binding site; (♦), NF-κB footprints at low-affinity sites. A schematic presentation of the nucleosomes is shown on the right side; the double headed arrow indicates the nucleosomal dyad. ( B ) UV laser footprinting of NF-κB bound to either naked 601_ D 7 -DNA (lanes 1 and 2) or to 601_D 7 -nucleosome (lanes 3 and 4) repeated with saturated amounts of NF-κB as indicated. ( C ) Same as (B), but for naked 601_D 0 DNA and 601_D 0 -nucleosomes. Note the absence of specific NF-κB footprint at the dyad in the case of the nucleosome.

    Journal: PLoS Genetics

    Article Title: Binding of NF-?B to Nucleosomes: Effect of Translational Positioning, Nucleosome Remodeling and Linker Histone H1

    doi: 10.1371/journal.pgen.1003830

    Figure Lengend Snippet: NF-κB binds specifically at the nucleosomal ends, but not at the nucleosomal dyad. ( A ) Upper panel : EMSA of NF-κB binding to naked 601_D 7 DNA (left) or to 601_D 7 nucleosome (right). Naked 32 P-end labeled 601_D 7 DNA or nucleosomes were incubated with increasing amount of NF-κB (1.5 fold serial dilution of NF-κB was performed starting with saturating amounts of 100 nM for naked DNA and 400 nM for nucleosome). The aliquots of the reaction mixtures were run on a 5% native PAGE. The positions of free DNA and nucleosomes are indicated, “cplx.” represents NF-κB – DNA/nuc complexes; lower panel: UV laser footprinting patterns of the NF-κB-DNA and NF-κB-nucleosomes complexes. The respective remaining mixtures were irradiated with a single 5 nanoseconds UV laser 266 nm pulse ( E pulse ∼0.1 J/cm 2 ), DNA was purified from the samples and then treated with Fpg glycosylase. The cleaved DNA fragments were separated on 8% sequencing gel and visualized by autoradiography; (*), NF-κB footprint at the high-affinity NF-κB binding site; (♦), NF-κB footprints at low-affinity sites. A schematic presentation of the nucleosomes is shown on the right side; the double headed arrow indicates the nucleosomal dyad. ( B ) UV laser footprinting of NF-κB bound to either naked 601_ D 7 -DNA (lanes 1 and 2) or to 601_D 7 -nucleosome (lanes 3 and 4) repeated with saturated amounts of NF-κB as indicated. ( C ) Same as (B), but for naked 601_D 0 DNA and 601_D 0 -nucleosomes. Note the absence of specific NF-κB footprint at the dyad in the case of the nucleosome.

    Article Snippet: The purified DNA was resuspended in resuspension buffer (10 mM Tris, pH 7.4, 30 mM NaCl, 1 mM EDTA, 1 mM DTT, 100 µg/ml BSA, 0.01% NP40) and cleaved with 0.1 units of either Fpg glycosylase or T4 endonuclease V (Trevigen) or both.

    Techniques: Binding Assay, Labeling, Incubation, Serial Dilution, Clear Native PAGE, Footprinting, Irradiation, Purification, Sequencing, Autoradiography

    Dilution driven H2A–H2B dimer eviction allows binding of NF-κB to Nucleosome Core Particle. ( A ) 152 bp DNA fragment derived from X. borealis somatic 5 S RNA gene containing single NF-κB site near the dyad NF1 (53–56) was amplified by PCR and uniquely 3′ end labeled with α- 32 P by Klenow. Nucleosomes were reconstituted on this labeled fragment as described previously. The DNA and nucleosomes at a concentration 40 nM were incubated with increasing amounts of NF-κB as indicated to allow the formation of stable complexes which were subsequently irradiated by a single high intensity UV laser pulse ( E pulse ∼0.1 J/cm 2 ). The formation of complexes was checked by EMSA (upper panel, DNA lane 1–3, nucleosomes lane 4–9), the positions of free DNA and nucleosomes are indicated, “cplx.” represents NF-κB – DNA/nuc complexes. The samples were split into two parts, DNA was purified and treated with Fpg glycosylase to cleave 8-oxoG (represented by ▸,lower panel, DNA lane 1–3 and nucleosome lane 4–9) and with T4 endonuclease V to cleave CPDs (represented by ◊, DNA lane 1′–3′ and nucleosome lane 4′–9′). The cleaved DNA fragments were visualized by 8% sequencing gel. ( B ) The same 152 bp 5S fragment was 5′ end labeled with γ- 32 P by T4 polynucleotide kinase and used for nucleosome reconstitution. DNA and nucleosomes, at 10 nM final concentration, were incubated with increasing amounts of NF-κB as indicated to allow the formation of complexes. The assembly of the complexes was checked by EMSA (upper panel, DNA lane 1–5, nucleosomes lane 1′–5′). The samples were irradiated with a single high intensity UV laser pulse ( E pulse ∼0.1 J/cm 2 ), treated with a mix of Fpg glycosylase and T4 endonuclease V to cleave both the 8-oxoG (▸) and CPDs (◊). Finally, the cleaved products were visualized by 8% sequencing gel (DNA, lane 1–5; nucleosomes lane 1′–5′). The NF-κB binding sites (vertical bold lines) and the NF-κB recognition sequences are shown. The arrows designated the nucleosomal dyad.

    Journal: PLoS Genetics

    Article Title: Binding of NF-?B to Nucleosomes: Effect of Translational Positioning, Nucleosome Remodeling and Linker Histone H1

    doi: 10.1371/journal.pgen.1003830

    Figure Lengend Snippet: Dilution driven H2A–H2B dimer eviction allows binding of NF-κB to Nucleosome Core Particle. ( A ) 152 bp DNA fragment derived from X. borealis somatic 5 S RNA gene containing single NF-κB site near the dyad NF1 (53–56) was amplified by PCR and uniquely 3′ end labeled with α- 32 P by Klenow. Nucleosomes were reconstituted on this labeled fragment as described previously. The DNA and nucleosomes at a concentration 40 nM were incubated with increasing amounts of NF-κB as indicated to allow the formation of stable complexes which were subsequently irradiated by a single high intensity UV laser pulse ( E pulse ∼0.1 J/cm 2 ). The formation of complexes was checked by EMSA (upper panel, DNA lane 1–3, nucleosomes lane 4–9), the positions of free DNA and nucleosomes are indicated, “cplx.” represents NF-κB – DNA/nuc complexes. The samples were split into two parts, DNA was purified and treated with Fpg glycosylase to cleave 8-oxoG (represented by ▸,lower panel, DNA lane 1–3 and nucleosome lane 4–9) and with T4 endonuclease V to cleave CPDs (represented by ◊, DNA lane 1′–3′ and nucleosome lane 4′–9′). The cleaved DNA fragments were visualized by 8% sequencing gel. ( B ) The same 152 bp 5S fragment was 5′ end labeled with γ- 32 P by T4 polynucleotide kinase and used for nucleosome reconstitution. DNA and nucleosomes, at 10 nM final concentration, were incubated with increasing amounts of NF-κB as indicated to allow the formation of complexes. The assembly of the complexes was checked by EMSA (upper panel, DNA lane 1–5, nucleosomes lane 1′–5′). The samples were irradiated with a single high intensity UV laser pulse ( E pulse ∼0.1 J/cm 2 ), treated with a mix of Fpg glycosylase and T4 endonuclease V to cleave both the 8-oxoG (▸) and CPDs (◊). Finally, the cleaved products were visualized by 8% sequencing gel (DNA, lane 1–5; nucleosomes lane 1′–5′). The NF-κB binding sites (vertical bold lines) and the NF-κB recognition sequences are shown. The arrows designated the nucleosomal dyad.

    Article Snippet: The purified DNA was resuspended in resuspension buffer (10 mM Tris, pH 7.4, 30 mM NaCl, 1 mM EDTA, 1 mM DTT, 100 µg/ml BSA, 0.01% NP40) and cleaved with 0.1 units of either Fpg glycosylase or T4 endonuclease V (Trevigen) or both.

    Techniques: Binding Assay, Derivative Assay, Amplification, Polymerase Chain Reaction, Labeling, Concentration Assay, Incubation, Irradiation, Purification, Sequencing