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t2aa 21921  (Cayman Chemical)


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    Cayman Chemical t2aa 21921
    Pharmacological inhibitors targeting both DNA replication DNA polymerase and DNA repair polymerase abolish or significantly reduce AAV2 DNA replication in vitro . (A) Purification of AAV2 Rep68 His protein. AAV2 Rep68 His protein was purified as described in Materials and Methods. The eluted protein from peaked fractions were separated on an SDS-(12%) PAGE gel, along with a protein size ladder (M), and stained with Coomassie brilliant blue. (B) In vitro DNA replication assay. Various inhibitors HAMNO (specifically targeting to RPA complex) at 100 μM, <t>T2AA</t> (to PCNA complex) at 1 μM, aphidicolin (to Pol α/δ/ε) at 10 μg/mL, MK886 (to Pol κ/η) at 200 μM, and PNR7-02 (to Pol η/λ) at 20 μM were added into the reaction of in vitro replication assays. DMSO served as a vehicle control. After DpnI digestion for 1 h at 37°C, the in vitro replicated products (lanes 2 to 7) were resolved on 1% agarose gel, and stained with ethidium bromide (EB) for DpnI digested inputs (lower panel). The dehydrated gel was exposed to a phosphor screen and scanned on a Typhoon FLA 9000 scanner. (B, lane 1), the reaction product without DpnI digestion was loaded as a size maker (~4.7 kb). The arrowhead indicates the in vitro replicated AAV2 genome. (C) Quantification of the relative replication efficiency. The fully in vitro replicated (DpnI-digestion resistant) viral DNA signals at ~4.7 kb were quantified by using ImageQuant Tl (IQTL) 8.2 software (Cytiva). The quantities are presented as relative levels to the DMSO control (B, lane 7). The data were obtained from triple experiments. ****, P < 0.0001.
    T2aa 21921, supplied by Cayman Chemical, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/t2aa+21921/pmc09973366-186-5-7?v=Cayman+Chemical
    Average 90 stars, based on 1 article reviews
    t2aa 21921 - by Bioz Stars, 2026-07
    90/100 stars

    Images

    1) Product Images from "Adeno-Associated Virus Monoinfection Induces a DNA Damage Response and DNA Repair That Contributes to Viral DNA Replication"

    Article Title: Adeno-Associated Virus Monoinfection Induces a DNA Damage Response and DNA Repair That Contributes to Viral DNA Replication

    Journal: mBio

    doi: 10.1128/mbio.03528-22

    Pharmacological inhibitors targeting both DNA replication DNA polymerase and DNA repair polymerase abolish or significantly reduce AAV2 DNA replication in vitro . (A) Purification of AAV2 Rep68 His protein. AAV2 Rep68 His protein was purified as described in Materials and Methods. The eluted protein from peaked fractions were separated on an SDS-(12%) PAGE gel, along with a protein size ladder (M), and stained with Coomassie brilliant blue. (B) In vitro DNA replication assay. Various inhibitors HAMNO (specifically targeting to RPA complex) at 100 μM, T2AA (to PCNA complex) at 1 μM, aphidicolin (to Pol α/δ/ε) at 10 μg/mL, MK886 (to Pol κ/η) at 200 μM, and PNR7-02 (to Pol η/λ) at 20 μM were added into the reaction of in vitro replication assays. DMSO served as a vehicle control. After DpnI digestion for 1 h at 37°C, the in vitro replicated products (lanes 2 to 7) were resolved on 1% agarose gel, and stained with ethidium bromide (EB) for DpnI digested inputs (lower panel). The dehydrated gel was exposed to a phosphor screen and scanned on a Typhoon FLA 9000 scanner. (B, lane 1), the reaction product without DpnI digestion was loaded as a size maker (~4.7 kb). The arrowhead indicates the in vitro replicated AAV2 genome. (C) Quantification of the relative replication efficiency. The fully in vitro replicated (DpnI-digestion resistant) viral DNA signals at ~4.7 kb were quantified by using ImageQuant Tl (IQTL) 8.2 software (Cytiva). The quantities are presented as relative levels to the DMSO control (B, lane 7). The data were obtained from triple experiments. ****, P < 0.0001.
    Figure Legend Snippet: Pharmacological inhibitors targeting both DNA replication DNA polymerase and DNA repair polymerase abolish or significantly reduce AAV2 DNA replication in vitro . (A) Purification of AAV2 Rep68 His protein. AAV2 Rep68 His protein was purified as described in Materials and Methods. The eluted protein from peaked fractions were separated on an SDS-(12%) PAGE gel, along with a protein size ladder (M), and stained with Coomassie brilliant blue. (B) In vitro DNA replication assay. Various inhibitors HAMNO (specifically targeting to RPA complex) at 100 μM, T2AA (to PCNA complex) at 1 μM, aphidicolin (to Pol α/δ/ε) at 10 μg/mL, MK886 (to Pol κ/η) at 200 μM, and PNR7-02 (to Pol η/λ) at 20 μM were added into the reaction of in vitro replication assays. DMSO served as a vehicle control. After DpnI digestion for 1 h at 37°C, the in vitro replicated products (lanes 2 to 7) were resolved on 1% agarose gel, and stained with ethidium bromide (EB) for DpnI digested inputs (lower panel). The dehydrated gel was exposed to a phosphor screen and scanned on a Typhoon FLA 9000 scanner. (B, lane 1), the reaction product without DpnI digestion was loaded as a size maker (~4.7 kb). The arrowhead indicates the in vitro replicated AAV2 genome. (C) Quantification of the relative replication efficiency. The fully in vitro replicated (DpnI-digestion resistant) viral DNA signals at ~4.7 kb were quantified by using ImageQuant Tl (IQTL) 8.2 software (Cytiva). The quantities are presented as relative levels to the DMSO control (B, lane 7). The data were obtained from triple experiments. ****, P < 0.0001.

    Techniques Used: In Vitro, Purification, Staining, Control, Agarose Gel Electrophoresis, Software



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    t2aa 21921  (Cayman Chemical)
    90
    Cayman Chemical t2aa 21921
    Pharmacological inhibitors targeting both DNA replication DNA polymerase and DNA repair polymerase abolish or significantly reduce AAV2 DNA replication in vitro . (A) Purification of AAV2 Rep68 His protein. AAV2 Rep68 His protein was purified as described in Materials and Methods. The eluted protein from peaked fractions were separated on an SDS-(12%) PAGE gel, along with a protein size ladder (M), and stained with Coomassie brilliant blue. (B) In vitro DNA replication assay. Various inhibitors HAMNO (specifically targeting to RPA complex) at 100 μM, <t>T2AA</t> (to PCNA complex) at 1 μM, aphidicolin (to Pol α/δ/ε) at 10 μg/mL, MK886 (to Pol κ/η) at 200 μM, and PNR7-02 (to Pol η/λ) at 20 μM were added into the reaction of in vitro replication assays. DMSO served as a vehicle control. After DpnI digestion for 1 h at 37°C, the in vitro replicated products (lanes 2 to 7) were resolved on 1% agarose gel, and stained with ethidium bromide (EB) for DpnI digested inputs (lower panel). The dehydrated gel was exposed to a phosphor screen and scanned on a Typhoon FLA 9000 scanner. (B, lane 1), the reaction product without DpnI digestion was loaded as a size maker (~4.7 kb). The arrowhead indicates the in vitro replicated AAV2 genome. (C) Quantification of the relative replication efficiency. The fully in vitro replicated (DpnI-digestion resistant) viral DNA signals at ~4.7 kb were quantified by using ImageQuant Tl (IQTL) 8.2 software (Cytiva). The quantities are presented as relative levels to the DMSO control (B, lane 7). The data were obtained from triple experiments. ****, P < 0.0001.
    T2aa 21921, supplied by Cayman Chemical, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/t2aa+21921/pmc09973366-186-5-7?v=Cayman+Chemical
    Average 90 stars, based on 1 article reviews
    t2aa 21921 - by Bioz Stars, 2026-07
    90/100 stars
    		  Buy from Supplier

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    Pharmacological inhibitors targeting both DNA replication DNA polymerase and DNA repair polymerase abolish or significantly reduce AAV2 DNA replication in vitro . (A) Purification of AAV2 Rep68 His protein. AAV2 Rep68 His protein was purified as described in Materials and Methods. The eluted protein from peaked fractions were separated on an SDS-(12%) PAGE gel, along with a protein size ladder (M), and stained with Coomassie brilliant blue. (B) In vitro DNA replication assay. Various inhibitors HAMNO (specifically targeting to RPA complex) at 100 μM, T2AA (to PCNA complex) at 1 μM, aphidicolin (to Pol α/δ/ε) at 10 μg/mL, MK886 (to Pol κ/η) at 200 μM, and PNR7-02 (to Pol η/λ) at 20 μM were added into the reaction of in vitro replication assays. DMSO served as a vehicle control. After DpnI digestion for 1 h at 37°C, the in vitro replicated products (lanes 2 to 7) were resolved on 1% agarose gel, and stained with ethidium bromide (EB) for DpnI digested inputs (lower panel). The dehydrated gel was exposed to a phosphor screen and scanned on a Typhoon FLA 9000 scanner. (B, lane 1), the reaction product without DpnI digestion was loaded as a size maker (~4.7 kb). The arrowhead indicates the in vitro replicated AAV2 genome. (C) Quantification of the relative replication efficiency. The fully in vitro replicated (DpnI-digestion resistant) viral DNA signals at ~4.7 kb were quantified by using ImageQuant Tl (IQTL) 8.2 software (Cytiva). The quantities are presented as relative levels to the DMSO control (B, lane 7). The data were obtained from triple experiments. ****, P < 0.0001.

    Journal: mBio

    Article Title: Adeno-Associated Virus Monoinfection Induces a DNA Damage Response and DNA Repair That Contributes to Viral DNA Replication

    doi: 10.1128/mbio.03528-22

    Figure Lengend Snippet: Pharmacological inhibitors targeting both DNA replication DNA polymerase and DNA repair polymerase abolish or significantly reduce AAV2 DNA replication in vitro . (A) Purification of AAV2 Rep68 His protein. AAV2 Rep68 His protein was purified as described in Materials and Methods. The eluted protein from peaked fractions were separated on an SDS-(12%) PAGE gel, along with a protein size ladder (M), and stained with Coomassie brilliant blue. (B) In vitro DNA replication assay. Various inhibitors HAMNO (specifically targeting to RPA complex) at 100 μM, T2AA (to PCNA complex) at 1 μM, aphidicolin (to Pol α/δ/ε) at 10 μg/mL, MK886 (to Pol κ/η) at 200 μM, and PNR7-02 (to Pol η/λ) at 20 μM were added into the reaction of in vitro replication assays. DMSO served as a vehicle control. After DpnI digestion for 1 h at 37°C, the in vitro replicated products (lanes 2 to 7) were resolved on 1% agarose gel, and stained with ethidium bromide (EB) for DpnI digested inputs (lower panel). The dehydrated gel was exposed to a phosphor screen and scanned on a Typhoon FLA 9000 scanner. (B, lane 1), the reaction product without DpnI digestion was loaded as a size maker (~4.7 kb). The arrowhead indicates the in vitro replicated AAV2 genome. (C) Quantification of the relative replication efficiency. The fully in vitro replicated (DpnI-digestion resistant) viral DNA signals at ~4.7 kb were quantified by using ImageQuant Tl (IQTL) 8.2 software (Cytiva). The quantities are presented as relative levels to the DMSO control (B, lane 7). The data were obtained from triple experiments. ****, P < 0.0001.

    Article Snippet: We used HAMNO (#S0148, Selleckchem), T2AA (#21921, Cayman Chemical), aphidicolin (#14007, Cayman Chemical), MK886 (#1311, Tocris Bioscience), and PNR7-02 (#2965, Axon Medchem).

    Techniques: In Vitro, Purification, Staining, Control, Agarose Gel Electrophoresis, Software