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  1. Link

    ds-dna-antibody-35i9-dna-bsa-and-azide-free-ab27156.pdf

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Anti-ds DNA antibody [35I9 DNA] - BSA and Azide free (ab27156)

  • Datasheet
Reviews (10)Q&A (10)References (71)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ds DNA antibody [35I9 DNA] - BSA and Azide free (ab27156)

    Key features and details

    • Mouse monoclonal [35I9 DNA] to ds DNA - BSA and Azide free
    • Suitable for: IHC-P
    • Reacts with: Species independent
    • Isotype: IgG2a

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    Overview

    • Product name

      Anti-ds DNA antibody [35I9 DNA] - BSA and Azide free
      See all ds DNA primary antibodies
    • Description

      Mouse monoclonal [35I9 DNA] to ds DNA - BSA and Azide free
    • Host species

      Mouse
    • Specificity

      Primarily Double stranded DNA. Measurements by immuno-CE yielded KD's of 0.71 µM and 0.09 µM, for the interaction of this antibody with ss- and dsDNA, respectively. Strong reactivity with both ss- and dsDNA has been observed on dotblots as well as very weak reactivity with RNA.
    • Tested applications

      Suitable for: IHC-Pmore details
    • Species reactivity

      Reacts with: Species independent
    • Immunogen

      The details of the immunogen for this antibody are not available.

    • Positive control

      • IHC-P: human skin tissue sections.
    • General notes

      This antibody has been shown to be useful, in the detection of dsDNA in fx. Crithidia luciliae -a monoflagellate protozoan,containing a giant mitochondrion. Very good reactivity both with dsDNA and ssDNA on NC-dotblots has been observed. The minimal size for DNA binding for this antibody is >16 bases and there is a inverse proportionality between binding and ionic strength.

      Abcam recommended secondaries - Goat Anti-Mouse HRP (ab205719) and Goat Anti-Mouse Alexa Fluor® 488 (ab150113).

      See other anti-mouse secondary antibodies that can be used with this antibody.

      The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. 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, along with publications, customer reviews and Q&As

    Properties

    • Form

      Liquid
    • Storage instructions

      Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
    • Storage buffer

      pH: 7.20
      Constituent: PBS
    • Carrier free

      Yes
    • Concentration information loading...
    • Purity

      Protein A purified
    • Primary antibody notes

      This antibody has been shown to be useful, using immunofluorescence, in the detection of dsDNA in fx. Crithidia luciliae -a monoflagellate protozoan,containing a giant mitochondrion. The antibody can be used for easy quantification of in vitro cell proliferation by ELISA and has successfully been used for monitoring cytotoxity and apoptosis as well as ELISA-based co-culture angiogenesis and proliferation assays. Very good reactivity both with dsDNA and ssDNA on NC-dotblots has been observed. The minimal size for DNA binding for this antibody is >16 bases and there is a inverse proportionality between binding and ionic strength.
    • Clonality

      Monoclonal
    • Clone number

      35I9 DNA
    • Myeloma

      x63-Ag8.653
    • Isotype

      IgG2a
    • Light chain type

      kappa
    • 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 IgG2a, kappa monoclonal [MG2a-53] - Isotype control (ab18415)

    Applications

    The Abpromise guarantee

    Our Abpromise guarantee covers the use of ab27156 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
    IHC-P (1)
    Use a concentration of 0.05 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
    Notes
    IHC-P
    Use a concentration of 0.05 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    Target

    • Relevance

      dsDNA (double stranded deoxyribonucleic acid) is the genetic material of all cells and many viruses and is a polymer of nucleotides. The monomer consists of phosphorylated 2-deoxyribose N-glycosidically linked to one of four bases, adenine, cytosine, guanine or thymine. These are linked together by 3',5'-phosphodiester bridges. In the Watson-Crick double-helix model, two complementary strands are wound in a right-handed helix and held together by hydrogen bonds between complementary base pairs.
    • Cellular localization

      Nuclear
    • Alternative names

      • Double stranded deoxyribonucleic acid antibody
      • Double stranded DNA antibody
      • dsDNA antibody

    Images

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ds DNA antibody [35I9 DNA] - BSA and Azide free (ab27156)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ds DNA antibody [35I9 DNA] - BSA and Azide free (ab27156)

       IHC image of ds DNA staining in a section of formalin-fixed paraffin-embedded normal human skin* 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 20mins. The section was then incubated with ab27156, 0.05ug/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. The inset secondary-only control image is taken from an identical assay without primary antibody.

      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

    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet download

      Download

    References (71)

    Publishing research using ab27156? Please let us know so that we can cite the reference in this datasheet.

    ab27156 has been referenced in 71 publications.

    • Shen JZ  et al. FBXO44 promotes DNA replication-coupled repetitive element silencing in cancer cells. Cell 184:352-369.e23 (2021). PubMed: 33357448
    • Sercel AJ  et al. Stable transplantation of human mitochondrial DNA by high-throughput, pressurized isolated mitochondrial delivery. Elife 10:N/A (2021). PubMed: 33438576
    • Hayden H  et al. ELISA detection of MPO-DNA complexes in human plasma is error-prone and yields limited information on neutrophil extracellular traps formed in vivo. PLoS One 16:e0250265 (2021). PubMed: 33886636
    • Sato A  et al. Immunofluorescence microscopy-based assessment of cytosolic DNA accumulation in mammalian cells. STAR Protoc 2:100488 (2021). PubMed: 34041502
    • Wan L  et al. Translation stress and collided ribosomes are co-activators of cGAS. Mol Cell 81:2808-2822.e10 (2021). PubMed: 34111399
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a question

    1-10 of 10 Q&A

    Question

    What concentration was this antibody used at in ELISA?

    Read More

    Abcam community

    Verified customer

    Asked on Mar 21 2014

    Answer


    ab27156 was validated in ELISA, coated with 20 ug/ml dsDNA in PBS. The antibody should be diluted from 0.5 to 2 µg/ml, depending on the batch.

    Read More

    Kevin Hanson

    Abcam Scientific Support

    Answered on Mar 21 2014

    Question

    I am looking for monoclonal antibody against ds DNA (I think it does not matter if it recognizes also ss DNA), that would be specific or at least would work with Staphylococcus aureus DNA for IF experiments. Thus I need antibody that will not bind to protein A. I would also need a secondary antibody coupled with a red fluorophore (against this DNA antibody). Could you give me some advice to choose the best antibodies for my experiment and eventually their price?

    Many thanks in advance for your help.

    Best Regards,

    Read More

    Abcam community

    Verified customer

    Asked on Oct 23 2012

    Answer

    Thank you for contacting us.

    Please check the following primary antibody datasheets

    https://www.abcam.com/ds-DNA-antibody-HYB331-01-ab27156.html

    https://www.abcam.com/ds-DNA-antibody-11B6-ab418.html

    The compatible secondary antibodies are;

    https://www.abcam.com/Goat-polyclonal-Secondary-Antibody-to-Mouse-IgG2a-heavy-chain-PE-Texas-Red-ab51410.html

    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 Oct 23 2012

    Question

    How much DNA can be detected using these antibodies? Do you have information on levels of detection? i.e. how much DNA can be detected per ug of antibody?

    Read More

    Abcam community

    Verified customer

    Asked on May 16 2012

    Answer

    Thank you for your inquiry and I appreciate your patience with this reply.
    For ab27156,the lab did nothave any data on how much DNA can be detected with the ds DNA antibody. However different specificities regarding the binding characteristics of the antibody are described below:
    Specificity: Primarily double stranded DNA (dsDNA). Measurements by immuno-CE yielded KD’s of 0.71 μM and 0.09 μM, for the interaction of ab27156 with ss- and dsDNA, respectively (1). Strong reactivity with both ss- and dsDNA has been observed on dot blots as well as very weak reactivity with RNA.
    Reactivity:In combination with immunofluorescence the antibody is well suited for detection of dsDNA in fx. Crithidia luciliae, a monoflagellate protozoa, containing a giant mitochondrion (2). The antibody can be used for easy quantification of in vitro cell proliferation by ELISA and has successfully been used for monitoring cytotoxity and apoptosis as well as ELISA-based co-culture angiogenesis and proliferation assays (3). Very good reactivity both with dsDNA and ssDNA on NC-dotblots has been observed. The minimal size for DNA bindingab27156 is >16 bases and there is an inverse proportionality between binding and ionic strength (1).
    1) N.H.H. Heegaard, D.T. Olsen and K.P. Larsen (1996) Journal of Chromatography A, 744: 285-294. (attached)
    2) J.B Peter and Y. Schoenfeld (1996) Autoantibodies, Elsevier. .230-236.
    3) T. Friis, B. Kjaer Sorensen, A.M. Engel, J. Rygaard and G. Houen (2003) APMIS. 111(6):658-68.
    For ab78856, the lab did not have any further data on the detection limits of the antibody andno information on the amount of DNA detected per ug of the antibody.
    I hope this information helps. Please contact us with any other questions.

    Read More

    Abcam Scientific Support

    Answered on May 16 2012

    Question

    Dear Abcam, One of customers is asking for antibodies against RNA and DNA for use at ELISA experiments. I realise that is odd question. Can you advise anything? Thank you,  

    Read More

    Abcam community

    Verified customer

    Asked on Nov 25 2011

    Answer

    Thank you for contacting us. Please check the datasheet for anti DNA antibodies ab27156; https://www.abcam.com/ds-DNA-antibody-HYB331-01-ab27156.html ab418; https://www.abcam.com/ds-DNA-antibody-11B6-ab418.html Please check the compatible secondary’s also ab8516 ab8517 ab8518 ab9175 ab97007 ab97008 ab97010 We unfortunately do not have anti RNA antibodies. Please select antibodies against RNA associated proteins.  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 Nov 25 2011

    Question

    I would like to use your antibody anti-ds DNA antibody [HYB331-01] to immunoprecipitate cross-linked shreaded DNA and perform a ligation reaction directly on the precipitation beads. - What does your antibody exactly recognizes? A specific sequence? Phosphodiester bonds? Can you tell me? - Would the interaction be maintained in a ligation buffer (typically 400 mM Tris-HCl, 100 mM MgCl2, 100 mM DTT, 5 mM ATP, pH 7.8 at 25°C)? - What is a good buffer to bind dsDNA?

    Read More

    Abcam community

    Verified customer

    Asked on Oct 06 2011

    Answer

    Thank you for contacting us. To answer you questions regarding ab27156: 1) Unfortunately, the details of the immunogen for this antibody are not available. Therefore, I do not know where the antibody binds and what the epitope would be. But we do know that the antibody is specific primarily to double stranded DNA. Measurements by immuno-CE yielded KD's of 0.71 µM and 0.09 µM, for the interaction of this antibody with ss- and dsDNA, respectively. Strong reactivity with both ss- and dsDNA has been observed on dotblots as well as very weak reactivity with RNA. The minimal size for DNA binding for this antibody is >16 bases and there is a inverse proportionality between binding and ionic strength. 2) IP has not been tested with this antibody yet. The ligation buffer contains 100 mM DTT which would be of concern as this would reduce the S-S bonds of the antibody and the antibody structure would fall apart (into heavy chains and light chains). The other buffer components should be fine. If you like to try out the antibody, you can take advantage of our testing discount program. For UNTESTED species and/or applications, we have established a testing discount program. Here is a brief description of how it works: The testing discount program is for customers who like to use an antibody/protein on an untested species/application. You would purchase the antibody at full price, test it and submit an Abreview with your data (positive or negative). On your next order you will receive a discount for ONE antibody at the full price (100%) of the antibody you have tested. The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount. 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 Oct 06 2011

    Question

    Can you please confirm that this antibody does recongize also poly dA - dT sequences? Thank you very much!

    Read More

    Abcam community

    Verified customer

    Asked on Aug 25 2011

    Answer

    Thank you for contacting us. I am happy to confirm that the laboratory answered that this antibody recognizes dsDNA independently of the sequence. Although the laboratory has not tested specifically the poly dA-dT composition, we will cover this application with our guarantee for you. Should you have any problems with this antibody recognizing this sequence, please contact us and we will do the necessary. I would like also to encourage you, to submit an Abreview with your results. Abreviews are very useful for sharing your experience with an antibody with other researchers and you will also get rewards for this. For more information, please visit www.abcam.com/abreviews Thank you very much. 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 Aug 25 2011

    Question

    Erica is looking for further information in regards to how this antibody binds and how 'strong' it will be or any information about the immunogen. There is very little information on the datasheet.

    Read More

    Abcam community

    Verified customer

    Asked on Sep 27 2007

    Answer

    Thank you for your enquiry. I can suggest having a look at the following references which I hope will be helpful to you. I 1) J.B Peter and Y. Schoenfeld (1996) Autoantibodies, Elsevier. 230-236. 2) T. Friis, B. Kjaer Sorensen, A.M. Engel, J. Rygaard and G. Houen (2003) A quantitative ELISA-based co-culture angiogenesis and cell proliferation assay. APMIS, 111(6):658-68. 3) N.H.H. Heegaard, D.T. Olsen and K.P. Larsen (1996) Immuno-capillary electrophoresis for the characterization of a monoclonal antibody against DNA. Journal of Chromatography A, 744: 285-294. This third reference states the following: 'A monoclonal antibody against DNA established from a mouse strain that spontaneously develops systemic lupus erythematosus was characterized by migration shift immuno-capillary electrophoresis. The minimal size for DNA binding antibody was >16 bases and the interaction with a double-stranded 32-met oligonucleotide was almost one order of magnitude stronger than the interaction with a single-stranded oligonucleotide. The binding was highly dependent on the ionic strength conditions with an increase in binding with a decrease in ionic strength. The estimate of the dissociation constant for the antibody binding of a single-stranded 32-met oligonucleotide was 0.62 μM at pH 7.90. This value was in good agreement with the value of 0.44 μM measured by an independent method using biosensor (surface plasmon resonance) technology.' I hope this information is useful. Should you have any further questions, please do not hesitate to contact us again.

    Read More

    Abcam Scientific Support

    Answered on Sep 27 2007

    Question

    Thanks so much for your quick response. You say that this antibody is not specific for prokaryotic DNA. By this do you mean that it has not been tested against it or it has been tested and does not work. It is interesting that in your data sheet on this product it does say that it has worked using immunofluorescence on a protozoan. while this is obviously not a prokaryote it is at least a lower eukaryote. So I was hoping that this antibody would have a higher chance of success against prokaryotic DNA than others I have looked at.

    Read More

    Abcam community

    Verified customer

    Asked on Nov 08 2006

    Answer

    Thank you for getting back to me. I am sorry but I do not have any further details other than this antibody has been tested and shown to work on lower eukaryotes as you indicate in addition to human cell lines. In theory there is no reason that this antibody should not work on both eukaryotic and prokaryotic forms of DNA. The only difference that I can see in the naked forms is that one is circular and the other is linear. I am sorry that I cannot be of more assistance on this occasion.

    Read More

    Abcam Scientific Support

    Answered on Nov 09 2006

    Question

    I am interested in your anti DNA antibody ab27156. I am trying to localise bacterial DNA at the electron microscope level. Do you have any information on whether this antibody is also specific for procaryotic DNA and whether it has been used in electron microscopic applications.

    Read More

    Abcam community

    Verified customer

    Asked on Nov 08 2006

    Answer

    Thank you for your enquiry. This antibody is not specific for prokaryotic DNA. I can tell you that the testing of this antibody was performed using immunocytochemistry using acetone fixed Hep2 and Hela cells. It has been shown to work using ELISA, Immunocytochemistry/Immunofluorescence and Western blot. To our knowledge it has not been tested and shown to work using electron microscopy. That is not to say that it does not work, just that it is yet to be tested.

    Read More

    Abcam Scientific Support

    Answered on Nov 08 2006

    Question

    I have a scientist who was enquiring about ds DNA antibodies- I gave him some information but he wanted me to contact Abcam for further information- please refer to his reply: I wonder what part of DNA it recognizes (true, DNA is DNA) and if it will work on LR white resin embedded thin section (70 -100nm thick). you could forward these questions to the company so I have a better and whole story?

    Read More

    Abcam community

    Verified customer

    Asked on Apr 19 2006

    Answer

    Thank you for your enquiry. I can tell you that the testing of this antibody was performed by immunohistochemistry using acetone fixed Hep2 and Hela cells. Unfortunately I do not have further details as to where this antibody has been applied on LR white resin embedded thin sections. The epitope of this antibody is dsDNA. Unfortunately I do not have specific information as to what within the DNA molecule it recognises other than that it recognises ds as opposed to ss DNA and requires a minimum of 10 bases (one complete helical turn) to recognise its epitope. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

    Read More

    Abcam Scientific Support

    Answered on Apr 19 2006

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