Overview

  • Product name
    Anti-Dengue Virus NS1 glycoprotein antibody [DN3]
    See all Dengue Virus NS1 glycoprotein primary antibodies
  • Description
    Mouse monoclonal [DN3] to Dengue Virus NS1 glycoprotein
  • Host species
    Mouse
  • Tested applications
    Suitable for: WB, ICC/IFmore details
  • Species reactivity
    Reacts with: Dengue virus 1, Dengue virus 2, Dengue virus 3, Dengue virus 4
  • Immunogen

    Full length native protein purified from Dengue Virus 2 (16681) infected supernatant.

  • Positive control
    • It is important to ensure that DEN4 infection is effective. DEN4 is notoriously difficult to grow to good titres and NS1 antigens will not be found if infection is not well established.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer
    Preservative: 0.065% Sodium azide
  • Purity
    Tissue culture supernatant
  • Clonality
    Monoclonal
  • Clone number
    DN3
  • Isotype
    IgG1
  • Light chain type
    kappa
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab41616 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 1/50 - 1/100. Use under non reducing condition. Detects a band of approximately 40, 80 kDa (predicted molecular weight: 40, 80 kDa). Block membrane with PBS containing 5% non-fat skimmed milk (PBS-SM) for 30 minutes. Wash membrane and probe with serum diluted in PBS-SM with 10 mM sodium azide by incubating overnight on a rocker at room temperature.
ICC/IF 1/5 - 1/20.

Target

  • Relevance
    NS1 is one of 7 Dengue Virus non-structural proteins which are thought to be involved in viral replication. NS1 exists as a monomer in its immature form but is rapidly processed in the endoplasmic reticulum to form a stable dimer. A small amount of NS1 remains associated with intracellular organelles where it is thought to be involved in viral replication. The rest of NS1 is found either associated with the plasma membrane or secreted as a soluble hexadimer. NS1 is essential for viral viability but its precise biological function is unknown. Antibodies raised in response to NS1 in viral infection can cross react with cell surface antigens on epithelial cells and platelets and this has been implicated in the development of Dengue Hemorrhagic fever.
  • Database links
    • Alternative names
      • Dengue NS1 antibody
      • Dengue Virus non structural protein 1 antibody

    Images

    • All lanes : Anti-Dengue Virus NS1 glycoprotein antibody [DN3] (ab41616)

      Lane 1 : Dengue Virus 1 infected C6/36 cell lysate (unheated)
      Lane 2 : Dengue Virus 1 infected C6/36 cell lysate (boiled)
      Lane 3 : Dengue Virus 2 (NGC) infected C6/36 cell lysate (unheated)
      Lane 4 : Dengue Virus 2 (NGC) infected C6/36 cell lysate (boiled)
      Lane 5 : Dengue Virus 2 (16681) infected C6/36 cell lysate (unheated)
      Lane 6 : Dengue Virus 2 (16681) infected C6/36 cell lysate (boiled)
      Lane 7 : Dengue Virus 3 infected C6/36 cell lysate (unheated)
      Lane 8 : Dengue Virus 3 infected C6/36 cell lysate (boiled)
      Lane 9 : Dengue Virus 4 infected C6/36 cell lysate (unheated)
      Lane 10 : Dengue Virus 4 infected C6/36 cell lysate (boiled)

      Predicted band size: 40, 80 kDa
      Observed band size: 40,80 kDa
      why is the actual band size different from the predicted?



      Recognises NS1 dimer (~80 kDa) in unheated samples and NS1 monomer (~40 kDa) when samples are boiled.
    • Dengue Virus 2 infected Vero cells stained according to protocol with ab41349 antibody against Dengue Virus envelope glycoprotein NS1 (green). Cell nuclei are blue.
    • ab41616 staining NS1 glycoprotein in Dengue Virus 2 infected Huh-7.5 cells by Immunocytochemistry/ Immunofluorescence.
      Huh-7.5 cells were plated on glass coverslips, infected with Dengue Virus 2, fixed in methanol, and blocked, and antibody was added in 1× PBS + 0.1% saponin. Antibodies used include FASN polyclonal, left image (ab3844) or NS1 monoclonal antibody, middle image (ab41616). Secondary antibodies were species-appropriate Alexa-Fluor 488 or 594 secondary antibody. Stained coverslips were mounted with DAPI, right merged image.

    References

    This product has been referenced in:
    • Lennemann NJ & Coyne CB Dengue and Zika viruses subvert reticulophagy by NS2B3-mediated cleavage of FAM134B. Autophagy 13:322-332 (2017). Read more (PubMed: 28102736) »
    • Cai H  et al. Cell-death-inducing DFFA-like Effector B Contributes to the Assembly of Hepatitis C Virus (HCV) Particles and Interacts with HCV NS5A. Sci Rep 6:27778 (2016). Read more (PubMed: 27282740) »
    See all 8 Publications for this product

    Customer reviews and Q&As

    1-10 of 29 Abreviews or Q&A

    Abcam has not validated the combination of species/application used in this Abreview.
    Application
    Flow Cytometry
    Sample
    Human Cell (HeLa at 62hpi with DV2)
    Permeabilization
    Yes - BD CytoFix/CytoPerm
    Gating Strategy
    single cells by FSC and SSC
    Specification
    HeLa at 62hpi with DV2
    Preparation
    Cell harvesting/tissue preparation method: spatula scraping
    Sample buffer: cold PBS
    Fixation
    BD CytoFix/CytoPerm

    Lee Gehrke

    Verified customer

    Submitted Oct 07 2013

    Answer

    Thank you for contacting us.

    DN3 is just a clone name. The antibody is reactive to all serotypes.

    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

    Answer

    Thank you for your message which has been forwarded to me as my colleague Anja is currently out of the office.

    We appreciate the feedback from the customer, thank you for sending this information.

    In order to obtain the discount, the customer will need to submit this informationan Abreview. They can do this by clicking on the Abreviews tab on the datasheet, then click on the link to 'submit an Abreview'.

    Please click the 'Submit and Abreview' link on the following page:

    https://www.abcam.com/Dengue-Virus-NS1-glycoprotein-antibody-DN3-ab41616-reviews.html

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

    https://www.abcam.com/abreviews

    I hope this will be helpful. If you have any further questions regarding submission of the Abreview, please do not hesitate to letme know.

    Read More
    Abcam has not validated the combination of species/application used in this Abreview.
    Application
    ELISA
    Sample
    Escherichia coli Recombinant protein (Dengue NS1)
    Specification
    Dengue NS1
    Blocking step
    BSA as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
    Type
    Indirect

    Abcam user community

    Verified customer

    Submitted Jul 26 2012

    Question

    Dear Sir/ Madam,
    We are trying to develop and ELISA for Dengue NS1 detection and having been working with your products in some of our previous and other present work, we have considered using your dengue products as well. However, during our preliminary experiments, we have noticed that the dengue NS1 antigen (ab64456) cannot be detected by either the anti-dengue NS1 antibody (ab41616) that you are also suggesting for other costumers in the technical support page, or the anti-histidine tag antibody (ab9136). This has led us to decide for a replacement of the antigen NS1 antigen (ab64456).
    Moving forward, we outsourced another dengue NS1 antigen from a different supplier and also an anti-dengue NS1 antibody which has been used in ELISA and WB applications for the detection of native NS1 antigens. (The antibody reacts with the 3rd and 4th conformational epitope of the Dengue NS1 based on the immunogen preparation.) We further tested the anti-dengue NS1 antibody (ab41616) using the new NS1 antigen and comparing with the new anti-dengue NS1 antibody as well. However, anti-dengue NS1 antibody (ab41616) still does not react with the new dengue NS1 antigen even at higher concentrations of 1:250. In comparison, the new antibody detected the dengue NS1 antigen adsorbed, and is also detected and validated by the anti-histidine tag antibody (ab9136).
    Since, it took a little time for us to outsource another dengue NS1 antigen to further test the reactivity of the anti-dengue NS1 antibody (ab41616) and perform evaluation, we are very much sorry for the delay but we would like to request for the possibility of replacement for the anti-dengue NS1 antibody (ab41616) into Anti-E.coli antibody (ab25823) that we would like to use for our other project.

    We are looking forward for a positive response from you. Thank you in advance.
    Kind regards,

    Read More
    Answer

    Thank you for taking the time to contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody.

    I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been successful.

    I would like to reassure you that this antibody is tested and covered by our guarantee for WB. Before deciding how to proceed, I would like to investigate this particular case further for you, and also obtain some further information for our quality records to determine what may be the cause of the problem.

    In order to do this, I have enclosed a questionnaire below. I would appreciate if you could complete this. It will help you put the information we require together very easily. I have completed the sections for which information has already been provided.

    It would also be beneficial if you are able toconfirm:

    1. What other samples have been tested with this antibody? Has it detected endogenous protein in any samples? I can suggest it would be beneficial to try this if possible.

    2. Has the protein been stained with comassie on an SDS gel? Is it intact and running at the correct molecular weight?

    In the event that a product is not functioning in the tested applications and species cited on the product data sheet (and the problem has been reported within 6 months of purchase), we will be pleased to provide a credit note, free of charge replacement or refund.

    Thank you for your time and cooperation. We look forward to receiving the completed questionnaire.


    Order Details
    Antibody code:

    Problem
    Choose: Non-specific band Multiple bands No signal or weak signal High background

    Lot number

    Purchase order number
    or preferably Abcam order number:



    General Information
    Antibody storage conditions (temperature/reconstitution etc)


    Description of the problem (high background, wrong band size, more bands, no band etc.)


    Sample (Species/Cell extract/Nuclear extract/Purified protein/Recombinant protein etc.)


    Sample preparation (Buffer/Protease inhibitors/Heating sample etc.)


    Amount of protein loaded


    Electrophoresis/Gel conditions (Reducing or Non-reducing gel, % of the gel etc.)


    Transfer and blocking conditions (Buffer/time period, Blocking agent etc.)


    Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)


    Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)


    Detection method (ECL, ECLPlus etc.)


    Positive and negative controls used (please specify)



    Optimization attempts (problem solving)
    How many times have you tried the Western?



    Have you run a "No Primary" control?
    Yes No

    Do you obtain the same results every time?
    Yes No
    e.g. are the background bands always in the same place?


    What steps have you altered?


    Additional Notes:


    Image:
    We would appreciate if you are able to provide an image (including molecular weight markers) which would help us to asess the results.

    Read More

    Answer

    Thank youand your customer very much for your interest in anti dengue virusNS1 antibodies.

    I can confirm that we have three in our catalog: ab41490, ab41616 and ab41623.

    To our knowledge, none of these antibodies has been tested in ELISA yet.

    Therefore, I can offer a discount off a future purchase if your customerbuysone of them now, tests it in ELISA and submits feedback to us in the form of an Abreview. It doesn’t matter whether the Abreview is positive or negative, we would just really like to receive your customer'sfeedback. The discount would be to the value ofthe antibodyand can be used for the next order of any product.

    If your customer is interested in this offer, please follow these steps:

    1. Reply to this e-mail to let me know that your customer ( please include theire-mail address)would like to proceed and confirm which one of these antibodiesyour customer would like to testin ELISA . I will then send a discount code. This code must be issued before purchasingthe antibodyso please wait for my reply before ordering.

    2. Purchasethe antibodyeither by phone, fax, or online (www.abcam.com).

    3. Test it in ELISA.

    4. Let us know the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out how to submit an Abreview, please visit: https://www.abcam.com/abreviews.

    5. After the review is submitted to us, the discount code becomes active. Simply place your customer's new order by phone, fax, or on the web and mention the discount code. The discount can be redeemed for any product ordered and the discount code is valid for 4 months after issue.

    We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even if teh antibody turns out to be unsuitable for ELISA, your customerwill still receive the discount ontheir next purchase after your Abreview has been submitted.

    Please let me know if you or your customer have any questions about this offer and I would be happy to help you further.

    The Terms and Conditions of this offer can be found at: www.abcam.com/collaborationdiscount.

    Read More

    Answer

    Thank youfor your inquiry.

    I can confirm that we do have three anti NS1 antibodies for Dengue virus in our catalog..

    Isotype control are not included and have to be purchased separately.

    ab41616 (Anti-Dengue Virus NS1 glycoprotein antibody [DN3]):

    https://www.abcam.com/index.html?datasheet=41616 (or use the following: https://www.abcam.com/index.html?datasheet=41616).

    Heaton NS et al. Dengue virus nonstructural protein 3 redistributes fatty acid synthase to sites of viral replication and increases cellular fatty acid synthesis. Proc Natl Acad Sci U S A 107:17345-50 (2010). ICC/IF. PubMed: 20855599

    Isotype control: ab18447 (Mouse IgG1, kappa monoclonal [MG1-45] - Isotype control)
    https://www.abcam.com/index.html?datasheet=18447 (or use the following: https://www.abcam.com/index.html?datasheet=18447).

    ab41623 (Anti-Dengue Virus NS1 glycoprotein antibody [DN2]):

    https://www.abcam.com/index.html?datasheet=41623 (or use the following: https://www.abcam.com/index.html?datasheet=41623).

    Smith KM et al. Structural mutants of dengue virus 2 transmembrane domains exhibit host-range phenotype. Virol J 8:289 (2011). IF. PubMed: 21658241

    Isotype control: ab18447 (Mouse IgG1, kappa monoclonal [MG1-45] - Isotype control)
    https://www.abcam.com/index.html?datasheet=18447 (or use the following: https://www.abcam.com/index.html?datasheet=18447).

    ab41490 (Anti-Dengue Virus NS1 glycoprotein antibody [DN1]):

    https://www.abcam.com/index.html?datasheet=41490 (or use the following: https://www.abcam.com/index.html?datasheet=41490).

    Currently we do not know of any references using this antibody.

    Isotype control: ab18449 (Mouse IgG2a, kappa [MOPC-173] (low endotoxin) - isotype control)
    https://www.abcam.com/index.html?datasheet=18449 (or use the following: https://www.abcam.com/index.html?datasheet=18449).

    We do not have set expiration dates for our products. Most antibodies are stable and can last for anywhere from a few months to several years if stored properly, so we strongly recommend that you follow the storage instructions on the datasheet for the antibody you purchased. These conditions will vary among our antibodies, therefore, it is important to verify the storage conditions for each of our products when you receive them. We guarantee all of our products to work for at leastsix monthfrom the date of purchase when stored correctly.

    For more information on antibody storage and stability, please visit our Antibody Storage Guide on our Protocols and Troubleshooting Tips page (https://www.abcam.com/index.html?pageconfig=popular_protocols).

    We do not sell directly to Pakistan. Please contact our trusted distributor for pricing information:

    Friends Scientific Corporation
    3-B Ali Continental,
    1-Mozang Road
    Lahore
    54000
    Pakistan

    Tel 4237236176 Fax 4237321500 Email mailto:friends1@wol.net.pk

    I hope this information is helpful and wish you good luck with your experiments.

    Read More

    Answer

    Gracias por tu respuesta.
    He consultado esta información con mis compañeros del laboratorio.
    La glicoproteína (ab64456) contiene toda la secuencia excepto el dominio transmembranal. Además contiene 2x6 His tags, uno localizado en el extremo N terminal y el otro en el C terminal.
    Respecto al anticuerpo (ab41616) me comentan que el dímero del virus Dengue NS1 está presente en las muestras sin calentar (forma sin reducir), y por efecto del calor se convierte al monómero. El epítopo es por tanto conformacional. En cualquier caso imagino que para llevar a cabo el ensayo ELISA no usáis muestras reducidas. Si tenéis la oportunidad os aconsejaría que probarais la combinación de proteína /anticuerpo mediante WB usando las muestras sin reducir, para comprobar si el anticuerpo realmente está detectando la proteína.
    El epítopo del anticuerpo no se ha mapeado, por tanto no puedo saber si reconoce el extremo N terminal o C terminal de la proteína donde se localizan los His tags.
    En cualquier caso, como no hay más Dengue Virus NS1 glicoproteínas en el catálogo, puedo mandaros de reemplazo otro primario contra la misma, esperando que esta vez si la reconozca. Estas son las alternativas:
    https://www.abcam.com/Dengue-Virus-NS1-glycoprotein-antibody-DN1-ab41490.html
    https://www.abcam.com/Dengue-Virus-NS1-glycoprotein-antibody-DN2-ab41623.html]
    Espero vuestra respuesta para saber cómo proceder y qué solución os podemos dar.

    Read More

    Answer

    Thank you for your inquiry.

    ab41616 is a mouse monoclonal IgG1 with a kappa light chain, un-conjugated.

    Therefore a isotype control is needed that is also un-conjugated and of the IgG1 Isotype (preferably with a kappa light chain). Here are three possible antibodies:

    ab18447
    https://www.abcam.com/index.html?datasheet=18447 (or use the following: https://www.abcam.com/index.html?datasheet=18447).

    ab18443
    https://www.abcam.com/index.html?datasheet=18443 (or use the following: https://www.abcam.com/index.html?datasheet=18443).

    ab91353
    https://www.abcam.com/index.html?datasheet=91353 (or use the following: https://www.abcam.com/index.html?datasheet=91353).

    I hope this information is helpful and wish your customer good luck with their research.

    Read More

    Question

    Dear Sir/Madam,
    We are trying to develop an ELISA for Dengue NS1 detection and since we have been been working for a long time with your products, we have decided to use your dengue products. We are in the process of validating our immunocapture system. The capture antibody we are using is a monoclonal antibody against DEN1-4 NS1 (ab41616) that we purchased together with theDengue NS1 glycoprotein (ab64456). We have already validated the presence of the mouse monoclonalantibody against DEN1-4 NS1 (ab41616) in our ELISA support using a Goat polyclonal Anti-mouse (ab20043). After looking at the citation for themouse monoclonalantibody against DEN1-4 NS1 we are expecting a good immunocapture. However on our initial tries, we cannot detect the presence of the dengue NS1 antigen. So we checked some details in the technical support and we found out that the NS1 antigen has a histidine tag which have been missing in the initial data sheet we had. And so we decided to purchase lasttime a unit of Anti-histidine tag HRP (ab1187) to check on theDengue NS1 glycoprotein (HisTag) (ab64456) but unfortunately even at 2 ug/ml concentration, we are not able to detect its presence. We are thinking of the possibility that the antigen we received may not have histidine tag as initially stated in the product sheet? but as you have guaranteed on our previous inquiry, the antigen is containing histidine tag. As loyal costumers of your company, we would like to ask you for the possibility of requesting replacement of the dengue NS1 glycoprotein (ab64456) that we have initially purchased with a different batch preferably to proceed with our experiments.
    We are looking forward in working further with your dengue products.
    Thank you for your usual costumer support.
    Regards,

    Read More
    Answer

    Thank you for contacting us.
    I am sorry to hear you are having problems with one of our products. I appreciate the details provided. I am afraid that product ab41616 is not guaranteed to work in ELISA. We have tested this antibody in WB and ICC, but we are not aware of its suitability for ELISA assays.
    As far as I understand you are performing an indirect ELISA, coating your plate with the antigen ab64456 and then capturing it with the monoclonal antibody, using ab20043 (secondary) to detect your primary, is that correct? It could well be the primary antibody is not recognising ab64456 as it may not work for this application.
    I can also confirm the peptide ab64456 is tagged with 6 histidines. It is also possible the antibody is not detecting this protein if the epitope it recognizes is masked with the histidine tags.
    If you wish to I can try to find out the epitope targeted by ab41616 and check whether it is comprised within the tag region.
    Regarding the failure of ab1187 (Anti-6X His tag® antibody (HRP)) to detect the glycoprotein, it could be due to the presence of sodium azide to preserve the protein. The antibody is linked to HRP and this enzyme is very sensitive to sodium azide.
    In any case, as all of our products are covered by our Abpromise guarantee, if you wish to receive a replacement for ab64456, I will be more than happy to send it to you.
    However, if you want to receive the same protein, you may still experience the same problems. In order to target this protein, I can try to find within our catalogue a primary against it that does not detect the epitope within the tagged sequence, or any other solution you may prefer.
    I look forward to receiving your reply.

    Read More

    1-10 of 29 Abreviews or Q&A

    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

    Sign up