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    dna2-antibody-ab96488.pdf

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Epigenetics and Nuclear Signaling DNA / RNA DNA Synthesis Other
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Anti-DNA2 antibody (ab96488)

  • Datasheet
Reviews (3)Q&A (1)References (22)

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Western blot - Anti-DNA2 antibody (ab96488)
  • Western blot - Anti-DNA2 antibody (ab96488)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DNA2 antibody (ab96488)

Key features and details

  • Rabbit polyclonal to DNA2
  • Suitable for: WB, IHC-P
  • Reacts with: Human
  • Isotype: IgG

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Overview

  • Product name

    Anti-DNA2 antibody
    See all DNA2 primary antibodies
  • Description

    Rabbit polyclonal to DNA2
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-Pmore details
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse, Rat
  • Immunogen

    Synthetic peptide corresponding to Human DNA2 aa 1000 to the C-terminus (C terminal) conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab42548)

  • Positive control

    • This antibody gave a positive signal in the following Human Whole Cell lysates; HEK293, HEK293 (Nuclear) and U2OS and FFPE human liver tissue sections.
  • General notes

    ab96488 is raised using the same immunogen as ab42439. Batches sold under ab96488 have passed QC testing in Western Blot. In our hands, this product did not work in ICC/IF. For further information please contact our Customer Services team.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

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: 0.02% Sodium azide
    Constituent: PBS

    Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

    • Epigenetics and Nuclear Signaling
    • DNA / RNA
    • DNA Synthesis
    • Other

Associated products

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)

Applications

Our Abpromise guarantee covers the use of ab96488 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
WB Use a concentration of 1 µg/ml. Detects a band of approximately 125 kDa (predicted molecular weight: 120 kDa).
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Target

  • Function

    May function in chromosomal DNA replication.
  • Sequence similarities

    Belongs to the DNA2/NAM7 helicase family.
  • Target information above from: UniProt accession P51530 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 1763 Human
    • Entrez Gene: 327762 Mouse
    • Entrez Gene: 309762 Rat
    • Omim: 601810 Human
    • SwissProt: P51530 Human
    • SwissProt: Q6ZQJ5 Mouse
    • SwissProt: D3ZG52 Rat
    • Unigene: 532446 Human
    • Unigene: 21492 Mouse
    see all
  • Alternative names

    • DNA replication ATP-dependent helicase-like homolog antibody
    • DNA replication helicase 2 homolog antibody
    • DNA replication helicase 2, yeast, homolog of antibody
    • DNA replication helicase 2-like antibody
    • DNA replication helicase/nuclease 2 antibody
    • DNA2 antibody
    • DNA2-like helicase antibody
    • DNA2L antibody
    • DNA2L_HUMAN antibody
    • HDNA2 antibody
    • KIAA0083 antibody
    • PEOA6 antibody
    • SCKL8 antibody
    see all

Images

  • Western blot - Anti-DNA2 antibody (ab96488)
    Western blot - Anti-DNA2 antibody (ab96488)
    All lanes : Anti-DNA2 antibody (ab96488) at 1 µg/ml

    Lane 1 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
    Lane 2 : HEK293 (Human embryonic kidney cell line) Nuclear Lysate
    Lane 3 : U2OS (Human osteosarcoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 120 kDa
    Observed band size: 125 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 16 kDa, 18 kDa, 210 kDa, 37 kDa, 38 kDa, 54 kDa, 60 kDa, 70 kDa, 76 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 10 minutes


    This antibody detects multiple bands in addition to the band of interest, reflecting low levels of expression of DNA2. siRNA knockdown experiments have demonstrated that this antibody detects a band at ~120 kDa corresponding to human DNA2 (please see image courtesy of Julien Duxin).
  • Western blot - Anti-DNA2 antibody (ab96488)
    Western blot - Anti-DNA2 antibody (ab96488)Image courtesy of Julien Duxin and Sheila Stewart
    All lanes : Anti-DNA2 antibody (ab96488)

    Lane 1 : U2OS cells plus control shRNA
    Lane 2 : U2OS cells plus shDNA2-1 shRNA
    Lane 3 : U2OS cells plus shDNA2-2 shRNA

    Secondary
    All lanes : Anti-rabbit at 1/3000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 120 kDa
    Observed band size: 120 kDa



    Western blot showing detection of DNA2 by ab42439. A 120 kDa band corresponding to DNA2 was detected in lysates of U2OS cells treated with control shRNA (lane 1) but not in those from knockdown cells treated with two different shRNAs for DNA2 (lanes 2 and 3). Following pre-blocking for 30 mins in 5% milk, PBS-Tween, the membrane was incubated overnight at 4°C with ab42439 at a dilution of 1/1000 in 5% milk, PBS-Tween. The membrane was then incubated with the secondary antibody at 1/3000 dilution in 5% milk, PBS-Tween for 2 hours at RT, and washed 3 x 10 mins with PBS-Tween prior to ECL detection.
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DNA2 antibody (ab96488)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DNA2 antibody (ab96488)
    IHC image of ab96488 staining in human liver carcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab96488, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

Protocols

  • Western blot protocols
  • Immunohistochemistry protocols

Click here to view the general protocols

Datasheets and documents

    • Datasheet
  • References (22)

    Publishing research using ab96488? Please let us know so that we can cite the reference in this datasheet.

    ab96488 has been referenced in 22 publications.

    • Yu H  et al. Global crotonylome reveals CDYL-regulated RPA1 crotonylation in homologous recombination-mediated DNA repair. Sci Adv 6:eaay4697 (2020). PubMed: 32201722
    • Mariotti L  et al. The iron-sulphur cluster in human DNA2 is required for all biochemical activities of DNA2. Commun Biol 3:322 (2020). PubMed: 32576938
    • Garzón J  et al. Human RIF1-Protein Phosphatase 1 Prevents Degradation and Breakage of Nascent DNA on Replication Stalling. Cell Rep 27:2558-2566.e4 (2019). PubMed: 31141682
    • Mason JM  et al. Non-enzymatic roles of human RAD51 at stalled replication forks. Nat Commun 10:4410 (2019). PubMed: 31562309
    • Panier S  et al. SLX4IP Antagonizes Promiscuous BLM Activity during ALT Maintenance. Mol Cell 76:27-43.e11 (2019). PubMed: 31447390
    View all Publications for this product

    Customer reviews and Q&As

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

    1-4 of 4 Abreviews or Q&A

    Immunoprecipitation abreview for Anti-DNA2 antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunoprecipitation
    Sample
    Human Cell lysate - whole cell (U2OS cells)
    Total protein in input
    500 µg
    Immuno-precipitation step
    Protein A/G
    Specification
    U2OS cells
    Read More

    Abcam user community

    Verified customer

    Submitted Jul 18 2017

    Western blot abreview for Anti-DNA2 antibody

    Below Average
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (U2OS cells)
    Gel Running Conditions
    Reduced Denaturing (4-12% Gradient Gel)
    Loading amount
    50 µg
    Specification
    U2OS cells
    Blocking step
    Licor Blocking Buffer as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    Submitted Oct 26 2016

    Question

    Dear Tech Support Team,   Kindly advise regarding ab96488:   ICC/IF abreview is shown in the abreviews sections but for some reason ICC/IF does not appear in the tested applications section in the website's datasheet. Will the customer get ABpromise for ICC/IF?   Thank you very much for your reply.   Have a nice day,

    Read More

    Abcam community

    Verified customer

    Asked on Sep 21 2011

    Answer

    Thank you for contacting us. Unfortunately, this antibody does not appear to be appropriate for ICC/IF. The Abreview showed poor results for this application and the in house testing has confirmed that the antibody should not be recommended for this application. This also means that it would not be covered by the Abpromise. We do not have any appropriate alternatives to offer from within our catalogue but you may be more successful in searching using biocompare: http://www.biocompare.com/ I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Read More

    Abcam Scientific Support

    Answered on Sep 21 2011

    Immunocytochemistry/ Immunofluorescence abreview for Anti-DNA2 antibody

    Poor
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (HeLa)
    Specification
    HeLa
    Fixative
    Paraformaldehyde
    Permeabilization
    Yes - 0.5% Triton X100
    Read More

    Dr. Kirk Mcmanus

    Verified customer

    Submitted Dec 09 2010

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com

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