FITC Anti-HIV1 tat antibody (ab43016)

Submit a review Q&A (8)References (2)
Flow Cytometry - FITC Anti-HIV1 tat antibody (ab43016)

    Key features and details

    • FITC Rabbit polyclonal to HIV1 tat
    • Suitable for: Flow Cyt
    • Conjugation: FITC. Ex: 493nm, Em: 528nm
    • Isotype: IgG

    Overview

    • Product name

      FITC Anti-HIV1 tat antibody
      See all HIV1 tat primary antibodies

    • Description

      FITC Rabbit polyclonal to HIV1 tat

    • Host species

      Rabbit

    • Conjugation

      FITC. Ex: 493nm, Em: 528nm

    • Specificity

      This antibody is specific for Tat protein and GST-Tat fusion protein.

    • Tested applications

      Suitable for: Flow Cytmore details

    • Species reactivity

      Reacts with: Other species

    • Immunogen

      Recombinant full length protein expressed in E. coli.

    • General notes

      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

    Applications

    The Abpromise guarantee

    Our Abpromise guarantee covers the use of ab43016 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
    Flow Cyt
    Notes
  • Application notes
    Flow Cyt: 1/200.
    For general use, dilute at 1:1 (V/V) in PBS.

    Not yet tested in other applications.
    Optimal dilutions/concentrations should be determined by the end user.

    • Target

    • Relevance

      HIV1 tat (Transactivator of Transcription) protein is a pleiotropic factor that induces a broad range of biological effects in numerous cell types. At the HIV promoter, tat is a powerful transactivator of gene expression which acts by inducing chromatin remodeling and by recruiting elongation-competent transcriptional complexes on to the viral LTR.

    • Cellular localization

      Nuclear. Upon stimulation with EDN1, it is exported from the nucleus to the perinuclear region.

    • Alternative names

      • p14 antibody
      • Protein Tat antibody
      • Transactivating regulatory protein antibody

    Images

    • Flow Cytometry - FITC Anti-HIV1 tat antibody (ab43016)
      Flow Cytometry - FITC Anti-HIV1 tat antibody (ab43016)
      ab43016 at a 1/200 dilution.

    References (2)

    Publishing research using ab43016? Please let us know so that we can cite the reference in this datasheet.

    ab43016 has been referenced in 2 publications.

    • Li H  et al. The glucocorticoid receptor-FKBP51 complex contributes to fear conditioning and posttraumatic stress disorder. J Clin Invest 130:877-889 (2020). PubMed: 31929189
    • Hategan A  et al. HIV Tat protein and amyloid-ß peptide form multifibrillar structures that cause neurotoxicity. Nat Struct Mol Biol 24:379-386 (2017). PubMed: 28218748

    Customer reviews and Q&As

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

    1-8 of 8 Abreviews or Q&A

    Question

    Your email has been very helpful! In your attachment, I see the antibody is most reactive at Tat 1 - 20, I assume that means amino acids 1 - 20? Because different publications refer to different amino acid numbering, just to be sure, would it be possible to have the amino acid sequence (or DNA sequence) to what 1 - 20 refers to? Given that I'm looking for Tat expression inside the cell...will all of these antibodies move inside the cell?
    Again, thank you so much for your help!

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    Answer


    Please find attached to this email theTAT sequence used for the epitope mapping. "TAT 1-20" corresponds to amino acids 1 to 20.

    Concerning the cell localisation, we recommend to permeabilise the cells with a standard protocol using Triton or another permeabilising detergent. For more details on Flow Cytometry intracellular staining I would recommend this link : https://www.abcam.com/index.html?pageconfig=resource&rid=11448

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    Question

    I would like to inquire what epitope/sequence this antibody binds on the Tat protein? I am interested in measuring the Tat levels in fixed ACH2 cells, which have a mutation at nucleotide xxx. I want to be sure that this antibody will work before purchasing. I saw a previous inquiry which mentioned it binds amino acids 1-20-- I was wondering if I could get a copy of the Tat sequence used for epitope mapping. I also wanted to inquire if this Tat sequence was from an HIV-1 subtype B or NL4-3 based Tat. Thank you in advance!

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    Answer

    Please find attached to this email the TAT sequence from clade B used for the epitope mapping, with reactivity scoring for different peptides. "TAT 1-20" corresponds to amino acids 1 to 20. Your nucleotide substitution is within that range and we do not know how that will affect reactivity.

    Here is the sequence used for the mapping:

    mepvdprlepwkhpgsqpktactncyckkccfhcqvcfitkalgisygrkkrrqrrrppqgsqthqvslskqptsqsrgdptgpke .

    The first 20 aa are identical in NL4-3 Tat.

    A comparison of the first 48 amino acids of Tat from several clades and NL4-3 is found in Figure 5 of the article at this link:

    http://www.ncbi.nlm.nih.gov/pmc/articles/PMC165250/

    J Virol. 2003 August; 77(15): 8602–8606. PMID: 12857933

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    Question

    hi
    I have included the original e-mail so that you can remember our previous communication.
    I have tested once the anti-TAT N3 clone (ab63957) antibody by Flow Cytometry in HEK293T transfected cells using an anti-FITC secondary antibody from DAKO but it did not work at all. Because the secondary antibody was out of date (expired but a PhD student was telling me that it is working) I have to test it with another secondary antibody that I am going to have this week. Thus I can have more clear and conclusive data by mid next week. Thus I will let you know and write an Abcam review.
    As you mention in this e-mail "I am also able to offer a testing discount for anti-Jun B (ab31421) and anti-Jun D (ab28837) to be tested in Flow Cytometry". Now I will be able to test both these antibodies by Flow Cytometry in several cell lines that I have confirmed for c-Jun and c-Fos. As thus what is the discount that you can offer me for both these products and can you generate a quote for these? May I order just one of them instead of both?
    Thank you very much in advance and best wishes

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    Answer

    Thank you for getting back in contact and letting me know of your progress.

    I am sorry to hear that ab63957 has as yet not produced the desired results using flow cytometry. I hope that using the new secondary antibody may provide you with better results. As mentioned in one of my previous emails, unfortunately we have not used this antibody in flow cytometry ourselves and have no reports as yet from any of our customers with how this antibody performs in this application. We cannot therefore guarantee that the antibody is suitable for this application. If you would like, I could have a look at the protocol you have been using in order to see if there are any suggestions I could make in order to improve the results observed so far. If you would like for me to do this could you please provide me with the protocol details by filling out the questionnaire I have attached to this email.

    I am please to hear that you may be interested in participating in the Abcam testing discount scheme to test ab31421 and ab28837 in flow cytometry. The offer I would be able to make is our current testing discount scheme which applies to antibodies in which the application or species you intend to use have not been specified on the datasheet of the antibody. This means that we do not currently have information on if the antibody is likely to work in that particular species or application and are therefore unable to guarantee that the antibody will work. In this case, neither ab31421or ab28837 have been tested inflow cytometry. This does not mean that they would not work in this application, but we simply do not have the information in order to guarantee it.

    This discount scheme works by you purchasing ab31421 and/or ab28837 (full price), and testing it in flow cytometry. Once you have tested the antibodies in this application and submitted the results through an Abreview (regardless of if the results are positive or negative) you are then entitled to a free primary antibody of your choice (or the equivalent of the original price off any product in our catalogue). The only restriction is that the antibody to be tested needs to be purchased, tested, the Abreview submitted and the free product claimed within 4 months.

    More information on this offer can be found from the following link:

    https://www.abcam.com/collaboratordiscount

    You can participate in this scheme with either both ab31421 and ab28837 or either one individually. Please do let meknow if you would liketo participate and with whichantibody as adiscount code needs to be generated before purchase of the antibody. Could you also please let me know which cell linesyou are intending touse? From which speciesdo the originate?

    I hope this information has been of help. If you require any further information please do not hesitate to contact us again.

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    Question

    Oui je souhaiterai recevoir un nouveau tube de ab43016 gratuit et je prendrai en compte vos recommandations d'utilisation.

    Merci.

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    Answer

    Merci pour votre réponse.

    Je me permets de vous transférer le numéro de commande de remplacement gratuit de ce produit : xx . Vous recevrez prochainement un mail de confirmation comprenant les détails d'expédition.

    J'espère que vous auriez des bons résultats avec le nouveau tube. N'hésitez pas à nous recontacter si vous avez des autres questions.

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    Question

    Bonjour,
    je vous contacte car j'ai récemment acheté un anticorps anti-Tat FITC, ab43016, et je rencontre quelques problèmes.
    Comme préconnisé dans la datasheet j'ai congelé l'anticorps à -20°C pour une longue conservation. Après une mise au point et quelques cycles de congélétion/décongélations j'ai obtenu un marquage sur une lignée Jurkat. Depuis j'essaie de reproduire ce résultat mais impossible, l'anticorps ne marque plus. Je suppose que les quelques cycles de congélétion/décongélations (environ 5) ont perturbé le produit. Si ceci est correcte c'est un peu décevant pour un anticorps acheté le 13 février! Que proposez-vous? Dans les guaranties il est stipulé que tous produits non performant seront remplacés, est-ce possible?
    xx

    Je vous remercie par avance de votre réponse.

    Cordialement,



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    Answer

    Merci de nous avoir contactés. Nous sommes désolés d'apprendre que le produit que vous avez reçu ne fonctionne pas comme attendu.

    Nous allons vous remplacer ce produit avec un nouveau tube. Par contre j'aimerai bien attirer votre attention sur le fait que nous recommandons d'éviter des cycles de congélation/décongélation (freeze/thaw cycles). Je vous conseille donc fortement, de faire des aliquots du nouveau tube, et une fois décongelée un des aliquots, le garder à 4 °C à l'abri de la lumière.

    Veuillez me confirmer que vous souhaitez recevoir un nouveau tube de ab43016.

    Merci de votre coopération. Je me réjouis de votre réponse.

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    Question

    Hello

    I am a PhD student in Australia, interested in the FITC-conjugated Tat antibody, and had a few questions (ab43016) (please excuse my naivety, I am quite new to using antibodies!)

    https://www.abcam.com/HIV1-tat-antibody-FITC-ab43016.html

    On your website, you mention it binds to full-length HIV Tat protein. How specific is this? Would it bind other proteins in the cell at all? Also, would this antibody bind to Tat that is made from just ORF1, or does it just bind full length? I am interested in detecting Tat protein from the first bit (before env in the genome), and do not expect full length Tat to be made. Where is the epitope on Tat for your antibody? One last question - if I expect to detect small amounts of my protein, what is the best mechanism to upscale the FACS results?

    Thank

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    Answer

    Thank you for your inquiry and your patience.

    Indeed, the questionsyou raised are very important. I have contacted the laboratory in order to find out more about this antibody.
    The antibody against Tat has been generated against the full length protein. However the laboratory has mapped the epitope and there is a high reactivity on the initial amino acids. Please see the attachment for the detailed results.

    The antibody specificity has been tested against
    - biotinilated Red Blood Cells linked to Tat recombinant protein
    - microspheres linked to Tat recombinant protein
    - Tat protein internalizing human macrophages
    - dendritic cells

    Flow cytometry expriments can be optimised by adjusting the blocking and the amount of antibody per target mainly. To upscale the antibody signal, I can recommend to use a biotinylated secondary antibody, followed by a fluorescent conjugated strepatvidin.The secondary antibodya as well as biotin-streptavidin will enhance the signal as well. I can recommend to have a look at our online flow cytometry protocols for further tips:
    https://www.abcam.com/index.html?pageconfig=resource&rid=11381
    https://www.abcam.com/index.html?pageconfig=resource&rid=11443
    https://www.abcam.com/index.html?pageconfig=popular_protocols

    I would like to reassure you also, that we do guarantee all our products to work as stated on the datasheet (https://www.abcam.com/abguarantee).

    I hope this information is helpful. Please do not hesitate to contact us again should you have any additional questions.

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    Question

    Dear Karin You may find it confusing with all these e-mails and as thus please disregard the two previous e-mails. In relation to ab63957 I am willing to accept the 50% discount that you ahve offered myself and order it this week on Wednesday. I am still willing to offer a review to Abcam in terms of performance of the antibody in Flow Cytometry analysis and obtain the same or another antibody in the future free of charge. However can you provide any discount for anti-Jun B (ab31421) and anti-Jun D (ab28837) to be tested by FACS-Flow Cytometry? My delivery address is: Thank you very much in advance and best wishes

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    Answer

    I'm sorry I was not available to deal with your query yesterday. I have generated the 50% discount for ab63957 as discussed which will be sent in a separate email. Let me know if you do not receive it. The testing discount code for this antibody is: 100%ABR-T7TY8 which expires on the 6th of August, 2012. In order to redeem this offer. Please order the antibody, test it in flow cytometry and tell us what you find through publishing an Abreview, putting the code given in the "Additional comments" section. Once this has been received you can place your order for the free primary antibody by placing your order in the usual way and quoting the discount code: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code. The terms and conditions of this offer can be found here: www.abcam.com/collaborationdiscount. I am also able to offer a testing discount for anti-Jun B (ab31421) and anti-Jun D (ab28837) to be tested in Flow Cytometry, could you just confirm with what species you intend to use these antibodies with and I will send them to you directly.

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    Question

    Dear xxxx I have tried the anti-TAT FITC antibody again (catalogue number: ab43016) at a dilution of 1/200 and 3/200 in much fewer cells (approxomately 5x105) staining overnight (>16h) and still it did not work very well. As thus and because I need an antibody immediately can you please sent me the replacement antibody:anti-HIV1 tat antibody [N3] (ab63957) that we have discussed over the phone? I would be really very grateful Thank you very much in advance and kind regards

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    Answer

    Thank you for letting me know of your progress. I am still waiting for a response as to the exact epitope recognised by this antibody, it may be as we discussed over the phone that the antibody is not able to recognise the portion of Tat-protein which you have taken to use as your fusion. Unfortunately as the datasheet states that the antibody recognises the full Tat protein I am unable to offer a free of charge replacement of your antibody. As I understand your disappointment and the effort you have gone to in order to try and make it work, I can offer as goodwill gesture a 50% discount on the alternative antibody ab63957. However, this antibody has not been tested in flow cytometry as yet and it cannot therefore be quarenteed to work I am also able to offer a testing discount. This will entitle you to a free primary antibody of your choice if you submit an abreview with the results of your experiments with ab63957(whether positive or negative). I am sorry that I could not be of more help in this instance. I look forward to hearing how you would like to proceed.

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