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Epigenetics and Nuclear Signaling DNA / RNA DNA / Nucleotides
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Anti-Thymine Dimer antibody [H3] (ab10347)

  • Datasheet
  • SDS
Reviews (7)Q&A (11)References (9)

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Immunocytochemistry/ Immunofluorescence - Anti-Thymine Dimer antibody [H3] (ab10347)

    Key features and details

    • Mouse monoclonal [H3] to Thymine Dimer
    • Suitable for: Southern Blot, ICC, Competitive ELISA, ELISA, ICC/IF
    • Reacts with: Species independent
    • Isotype: IgG1

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    Overview

    • Product name

      Anti-Thymine Dimer antibody [H3]
    • Description

      Mouse monoclonal [H3] to Thymine Dimer
    • Host species

      Mouse
    • Tested applications

      Suitable for: Southern Blot, ICC, Competitive ELISA, ELISA, ICC/IFmore details
    • Species reactivity

      Reacts with: Species independent
    • Immunogen

      Chemical/ Small Molecule.

    • Positive control

      • ICC/IF: HeLa cells.
    • General notes


      Non-radioactive labeling of DNA is typically based on the enzymatic incorporation of modified nucleotides, carrying a small chemical moiety such as biotin, digoxigenin or fluorescein. These tags are subsequently detected by specific reagents such as streptavidin or a specific antibody coupled to a signal-producing enzyme. Although very efficient and reliable, labeling by in vitro polymerization is time-consuming, expensive, and may require various post-label purification steps to remove an excess of unincorporated precursors. An alternative strategy for DNA labeling, is based on the UV-induced formation of cyclobutane thymine dimers. Several methods have been described for the detection of thymine dimers, which are based on chromato-graphic analysis, and on biochemical analysis with endonucleases specific for UV-irradiated DNA. In addition, methods utilizing antibodies specific for pyrimidine dimers and other UV-induced DNA lesions have evolved, which permit the study of the induction and repair of these lesions without the requirement of in vivo radiolabeling of DNA. Photoimmunodetection, is a rapid, reliable and low-cost supplement to existing methods for nonradioactive DNA labeling. It enables a sensitive and non-radioactive method for labeling, detection, and quantification of high molecular weight (HMW) DNA fragments. The method is based on the introduction of thymine dimers into DNA after separa-tion by pulse field gel electrophoresis (PFGE), followed by detection with thymine dimer specific antibodies. The method does not require any enzymatic or chemical manipulation of the DNA sample. Monoclonal anti-bodies reacting specifically with thymine dimer, facilitate investigations on the apoptotic process and the role of UV-induced pyrimidine dimers in the process of photocarcinogenesis.

    Properties

    • Form

      Liquid
    • Storage instructions

      Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
    • Storage buffer

      pH: 7.40
      Preservative: 0.097% Sodium azide
      Constituent: PBS
    • Concentration information loading...
    • Purity

      Protein G purified
    • Primary antibody notes

      Non-radioactive labeling of DNA is typically based on the enzymatic incorporation of modified nucleotides, carrying a small chemical moiety such as biotin, digoxigenin or fluorescein. These tags are subsequently detected by specific reagents such as streptavidin or a specific antibody coupled to a signal-producing enzyme. Although very efficient and reliable, labeling by in vitro polymerization is time-consuming, expensive, and may require various post-label purification steps to remove an excess of unincorporated precursors. An alternative strategy for DNA labeling, is based on the UV-induced formation of cyclobutane thymine dimers. Several methods have been described for the detection of thymine dimers, which are based on chromato-graphic analysis, and on biochemical analysis with endonucleases specific for UV-irradiated DNA. In addition, methods utilizing antibodies specific for pyrimidine dimers and other UV-induced DNA lesions have evolved, which permit the study of the induction and repair of these lesions without the requirement of in vivo radiolabeling of DNA. Photoimmunodetection, is a rapid, reliable and low-cost supplement to existing methods for nonradioactive DNA labeling. It enables a sensitive and non-radioactive method for labeling, detection, and quantification of high molecular weight (HMW) DNA fragments. The method is based on the introduction of thymine dimers into DNA after separa-tion by pulse field gel electrophoresis (PFGE), followed by detection with thymine dimer specific antibodies. The method does not require any enzymatic or chemical manipulation of the DNA sample. Monoclonal anti-bodies reacting specifically with thymine dimer, facilitate investigations on the apoptotic process and the role of UV-induced pyrimidine dimers in the process of photocarcinogenesis.
    • Clonality

      Monoclonal
    • Clone number

      H3
    • Isotype

      IgG1
    • Research areas

      • Epigenetics and Nuclear Signaling
      • DNA / RNA
      • DNA / Nucleotides

    Associated products

    • Compatible Secondaries

      • Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
      • Goat Anti-Mouse IgG H&L (HRP) (ab205719)
    • Isotype control

      • Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control (ab170190)

    Applications

    The Abpromise guarantee

    Our Abpromise guarantee covers the use of ab10347 in the following tested applications.

    The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

    Application Abreviews Notes
    Southern Blot
    Use a concentration of 0.5 - 1 µg/ml.
    ICC
    Use at an assay dependent dilution.
    Competitive ELISA
    Use at an assay dependent dilution.
    ELISA (1)
    Use at an assay dependent dilution.
    ICC/IF (2)
    Use at an assay dependent concentration.
    Notes
    Southern Blot
    Use a concentration of 0.5 - 1 µg/ml.
    ICC
    Use at an assay dependent dilution.
    Competitive ELISA
    Use at an assay dependent dilution.
    ELISA
    Use at an assay dependent dilution.
    ICC/IF
    Use at an assay dependent concentration.

    Images

    • Immunocytochemistry/ Immunofluorescence - Anti-Thymine Dimer antibody [H3] (ab10347)
      Immunocytochemistry/ Immunofluorescence - Anti-Thymine Dimer antibody [H3] (ab10347)This image is courtesy of an anonymous Abreview.
      ab10347 staining Thymine Dimer in HeLa cells by Immunocytochemistry/ Immunofluorescence.
      Cells were fixed in formaldehyde, permabilized using 0.5% Triton X-100, blocked with 5% BSA for 15 minutes at 20°C, then incubated with ab10347 at a 1/250 dilution for 16 hours at 4°C. The secondary used was an Alexa-Fluor 488 conjugated rabbit anti mouse polyclonal, used at a 1/500 dilution.

      See Abreview

    Protocols

    • Immunocytochemistry & immunofluorescence protocols

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet
    • SDS
  • References (9)

    Publishing research using ab10347? Please let us know so that we can cite the reference in this datasheet.

    ab10347 has been referenced in 9 publications.

    • Tedeschi F  et al. EFFECTOR OF TRANSCRIPTION factors are novel plant-specific regulators associated with genomic DNA methylation in Arabidopsis. New Phytol 221:261-278 (2019). PubMed: 30252137
    • Cobb AM  et al. Disruption of PCNA-lamins A/C interactions by prelamin A induces DNA replication fork stalling. Nucleus 7:498-511 (2016). PubMed: 27676213
    • Torregrosa-Muñumer R  et al. Low doses of ultraviolet radiation and oxidative damage induce dramatic accumulation of mitochondrial DNA replication intermediates, fork regression, and replication initiation shift. Mol Biol Cell 26:4197-208 (2015). PubMed: 26399294
    • Deng Y  et al. A sunblock based on bioadhesive nanoparticles. Nat Mater 14:1278-85 (2015). PubMed: 26413985
    • Anda S  et al. Cell-cycle analyses using thymidine analogues in fission yeast. PLoS One 9:e88629 (2014). PubMed: 24551125
    • Dantas TJ  et al. Defective nucleotide excision repair with normal centrosome structures and functions in the absence of all vertebrate centrins. J Cell Biol 193:307-18 (2011). WB . PubMed: 21482720
    • Myakishev-Rempel M  et al. Investigation of the peak action wavelength of light-activated gene transduction. Gene Ther 18:1043-51 (2011). PubMed: 21490685
    • Moriel-Carretero M & Aguilera A A Postincision-Deficient TFIIH Causes Replication Fork Breakage and Uncovers Alternative Rad51- or Pol32-Mediated Restart Mechanisms. Mol Cell 37:690-701 (2010). PubMed: 20227372
    • Parro V  et al. SOLID2: an antibody array-based life-detector instrument in a Mars Drilling Simulation Experiment (MARTE). Astrobiology 8:987-99 (2008). ELISA . PubMed: 19105755

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-10 of 18 Abreviews or Q&A

    Other abreview for Anti-Thymine Dimer antibody [H3]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Other - Do not use
    Sample
    Human Cell lysate - nuclear (WI38 cell line)
    Read More

    Abcam user community

    Verified customer

    Submitted Dec 08 2017

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Thymine Dimer antibody [H3]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (Human skin equivalent model, with and without UVB-)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: pH 8.0 EDTA buffer (90¯C/40min)
    Permeabilization
    No
    Specification
    Human skin equivalent model, with and without UVB-
    Blocking step
    Biocare Medical Background SNIPER as blocking agent for 10 minute(s) · Concentration: 100% · Temperature: 24°C
    Fixative
    10% NBF
    Read More

    Abcam user community

    Verified customer

    Submitted Jan 04 2017

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Thymine Dimer antibody [H3]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
    Sample
    Human Cell (U2OS)
    Specification
    U2OS
    Permeabilization
    Yes - triton-X 100
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Oct 21 2014

    ELISA abreview for Anti-Thymine Dimer antibody [H3]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    ELISA
    Sample
    Human Cell (u2os)
    Specification
    u2os
    Type
    Direct
    Blocking step
    Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
    Read More

    Dr. Andrew Cobb

    Verified customer

    Submitted Jul 16 2013

    Question

    Dear Tech support

    The datasheet for product ab10347 doesn't mention species reactivity.
    Is the antibody expected to react with Pig derived samples? Could you offer the Abcam guarantee for pig IC/IFF with this antibody? Do you think IHC-P would work as well? could this application be guaranteed?

    Thank you very much for your kind support and have a great week.
    Regards,

    Read More

    Abcam community

    Verified customer

    Asked on Oct 15 2012

    Answer

    Thank you for your message.

    I can confirm that Thymine Dimer is not a species specific target, and so this antibody should work in pig.

    As far as we are aware, ab10347 has never been tested in IHC-P. All tested applications covered by the 6 month guarantee are specified on our individual datasheets, and these are updated as soon as any new information is brought to our attention.

    If the customer would like to test ab10347 in IHC-P, please do not hesitate to contact me again prior to the purchase by replying to this message as they may be eligible for our testing discount program.

    Otherwise, we like to encourage all of our customers to submit an Abreview via the online product datasheet. We always appreciate customer feedback, whether positive or negative, and we make all product information available to researchers. Plus, each Abreview earns Abpoints that can be used for discounts on future purchases or rewards such as Amazon.com gift certificates.

    To find out more about our Abreview system, please see the following link:

    https://www.abcam.com/abreviews

    I hope this information is helpful. Please do not hesitate to contact me for any further advice or information.

    Read More

    Abcam Scientific Support

    Answered on Oct 15 2012

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Thymine Dimer antibody [H3]

    Below Average
    Abreviews
    Abreviews
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (Whole skin)
    Specification
    Whole skin
    Fixative
    Formaldehyde
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citrate
    Permeabilization
    Yes - 0.1% Tween-20
    Blocking step
    Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    Submitted Oct 11 2012

    Question

    1) Abcam product code ab 10347
    2) Abcam order reference number or product batch number : lot
    3) Description of the problem : Unable to see any positive staining of thymine dimers.
    4) Sample preparation:
    Species Mouse (Balb/c)
    Type of sample: Fresh frozen sections, perfusion fixed frozen sections, PFA/formalin fixed paraffin embedded sections, cells in culture, other: formalin fixed paraffin embedded sections
    Sample preparation
    Positive control: Untreated skin irradiated for 30 min with 8Kj/m2 UVB- Collected 30 min and 24 hour post irradiation
    5) Fixation step
    Yes/No
    If yes: Fixative agent and concentration : 10% buffered formalin
    Fixation time: 24 H
    Fixation temperature 4C
    *also tried unfixed (fresh), and acetone/methanol fixed*
    6) Antigen retrieval method : Sodium Citrate Buffer (10mM Sodium Citrate, 0.05% Tween 20, pH 6.0)- 30 min @ 95C (in steamer).
    Also tried tris-EDTA Buffer (10mM Tris Base, 1mM EDTA Solution, 0.05% Tween 20, pH 9.0
    7) Permeabilization method:
    Did you do a permeabilization step (details please) or add permeabilizing agent in any dilution buffers?
    Permeabilizing agent and concentration: Tried both triton X-100 (0.05%) and tween 20 (0.1%) in PBS: used in wash buffers
    8) Blocking agent (eg BSA, serum…): Tried both goat serum and horse serum at 10%
    Concentration: 10%
    Blocking time 1 hour, 2 hours, overnight Blocking temperature: 1 hour and 2 hours: RT; overnight: 4C
    9) Endogenous peroxidases blocked? Yes
    Endogenous biotins blocked? Yes (dako kit)
    10) Primary antibody (If more than one was used, describe in “additional notes”) :
    Concentration or dilution : Tried, 1:50, 1:100, 1:400, 1:1000, 1:2000, 1:4000
    Diluent buffer : Tried 1X TBS (10X TBS= (0.5M Tris Base, 9% NaCl, pH 8.4) ) with 10% serum. Also tried a PBS, 2mM EDTA, 1% BSA diluent.
    Incubation time: 1 hour, 2 hours, 3 hours, overnight
    11) Secondary antibody:
    Species: Used DAKO animal research kit to biotinylate primary antibody prior to application to tissue. Secondary used was peroxidise HRP
    Reacts against: biotin
    Concentration or dilution : as per kit
    Diluent buffer : as per kit
    Incubation time : 30 min, 1 hour, 2 hours
    Fluorochrome or enzyme conjugate: DAB chromagen/ substrate, as per kit.
    12) Washing after primary and secondary antibodies:
    Buffer: TBST
    Number of washes 3-5x 2-5 min washes (tried both)
    13) Detection method: Nikon inverted microscope, light microscopy
    14) How many times have you run this staining? 7 times
    Do you obtain the same results every time? Yes: no staining at all

    Read More

    Abcam community

    Verified customer

    Asked on Sep 27 2012

    Answer

    Thank you for taking time to complete our questionnaire and for contacting us. I am sorry to hear this antibody is not providing satisfactory results.

    Having reviewed the protocol details, and after all the optimisation attempts, I am afraid there are no protocol tips I could suggest to obtain better results from this antibody in IHC-P. It may well be this antibody does not work in this application.

    The discount code was issued for testing the antibody with mouse samples in ICC. Have you had the opportunity to test the antibody in these conditions?


    I hope this information is helpful. Please do not hesitate to contact us for further assistance.

    Read More

    Abcam Scientific Support

    Answered on Sep 27 2012

    Question

    These codes below are not working. I need to place an order today using them. Can you solve this problem? Thanks!

    Read More

    Abcam community

    Verified customer

    Asked on Jul 31 2012

    Answer

    I am sending you the new discount codes below, but would suggest if you encounter problems with using the codes for an online order, please give us a call so thatCustomer Service can help you with applying the codes to the order.

    DISCOUNT CODE: xxx for ab10347
    DISCOUNT CODE: xxx for ab23722
    EXPIRATION: xxx

    Please let me know if I can help you with anything else.

    Read More

    Abcam Scientific Support

    Answered on Jul 31 2012

    Immunohistochemistry (Frozen sections) abreview for Anti-Thymine Dimer antibody [H3]

    Average
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Human Tissue sections (Skin)
    Specification
    Skin
    Fixative
    Acetone
    Permeabilization
    No
    Read More

    Abcam user community

    Verified customer

    Submitted Jul 06 2012

    Question

    I found the original e-mails with the codes and they expired 1 day after I received them . Could you extend the expiration date for these codes? Thanks

    Read More

    Abcam community

    Verified customer

    Asked on Jul 06 2012

    Answer

    Thank you for your reply.

    I apologize for this oversight on our end, and I am resending you new codes that should work for you.

    DISCOUNT CODE: xxx for ab10347
    DISCOUNT CODE: xxx for ab23722
    EXPIRATION: xxx

    Thank you so much for submitting your Abreviews and testing these antibodies.

    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Use our products? Submit an Abreview. Earn rewards!
    https://www.abcam.com/abreviews

    Read More

    Abcam Scientific Support

    Answered on Jul 06 2012

    1-10 of 18 Abreviews or Q&A

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