Overview

  • Product name
    Anti-Desmoplakin I+II antibody [DP2.15]
    See all Desmoplakin I+II primary antibodies
  • Description
    Mouse monoclonal [DP2.15] to Desmoplakin I+II
  • Host species
    Mouse
  • Tested applications
    Suitable for: ICC, ICC/IF, WB, IHC-Frmore details
    Unsuitable for: IHC-P
  • Species reactivity
    Reacts with: Mouse, Rat, Chicken, Cow, Human
  • Immunogen

    Full length protein (Cow).

  • Positive control
    • Positive cell lines: Human A432 and MCF-7; Rat MH1C1; Bovine MDBK and BMGE. Stratified epithelia, simple epithelia, including glands, urothelium, thymic reticular epithelium, hepatocytes, intercalated disks of myocardium and arachnoid cells of meninges.
  • General notes
    Suitable for use in the detection of primary and metastatic carcinomas.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Add glycerol to a final volume of 50% for extra stability and aliquot. Store at -20°C. Avoid freeze / thaw cycle.
  • Storage buffer
    Preservative: 0.1% Sodium Azide
    Constituents: 0.5% BSA, PBS, pH 7.4
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Clonality
    Monoclonal
  • Clone number
    DP2.15
  • Isotype
    IgG1
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab16434 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
ICC 1/200.
ICC/IF 1/100.
WB Use at an assay dependent concentration. Predicted molecular weight: 331 kDa.
IHC-Fr 1/10.
  • Application notes
    Is unsuitable for IHC-P.
  • Target

    • Function
      Major high molecular weight protein of desmosomes. Involved in the organization of the desmosomal cadherin-plakoglobin complexes into discrete plasma membrane domains and in the anchoring of intermediate filaments to the desmosomes.
    • Tissue specificity
      Isoform DPI is apparently an obligate constituent of all desmosomes. Isoform DPII resides predominantly in tissues and cells of stratified origin.
    • Involvement in disease
      Keratoderma, palmoplantar, striate 2
      Cardiomyopathy, dilated, with woolly hair and keratoderma
      Arrhythmogenic right ventricular dysplasia, familial, 8
      Skin fragility-woolly hair syndrome
      Epidermolysis bullosa, lethal acantholytic
      Cardiomyopathy, dilated, with woolly hair, keratoderma, and tooth agenesis
    • Sequence similarities
      Belongs to the plakin or cytolinker family.
      Contains 17 plectin repeats.
      Contains 1 SH3 domain.
      Contains 6 spectrin repeats.
    • Domain
      Its association with epidermal and simple keratins is dependent on the tertiary structure induced by heterodimerization of these intermediate filaments proteins and most likely involves recognition sites located in the rod domain of these keratins.
      The N-terminal region is required for localization to the desmosomal plaque and interacts with the N-terminal region of plakophilin 1.
      The three tandem plakin repeat regions in the C-terminus mediate binding to intermediate filaments.
    • Post-translational
      modifications
      Ser-2849 is probably phosphorylated by a cAMP-dependent protein kinase. Phosphorylation on Ser-2849 probably affects its association with epidermal, simple cytokeratins and VIM intermediate filaments.
      Substrate of transglutaminase. Some glutamines and lysines are cross-linked to other desmoplakin molecules, to other proteins such as keratin, envoplakin, periplakin and involucrin, and to lipids like omega-hydroxyceramide (PubMed:9651377).
    • Cellular localization
      Cell junction, desmosome. Cytoplasm, cytoskeleton. Cell membrane. Innermost portion of the desmosomal plaque. Colocalizes with epidermal KRT5-KRT14 and simple KRT8-KRT18 keratins and VIM intermediate filaments network (PubMed:12802069). Localizes at the intercalated disk in cardiomyocytes (By similarity).
    • Information by UniProt
    • Database links
    • Alternative names
      • 250/210 kDa paraneoplastic pemphigus antigen antibody
      • DCWHKTA antibody
      • Desmoplakin (DPI DPII) antibody
      • Desmoplakin antibody
      • Desmoplakin I antibody
      • Desmoplakin II antibody
      • DESP_HUMAN antibody
      • DP antibody
      • DP I antibody
      • DP II antibody
      • DPI antibody
      • DPII antibody
      • DSP antibody
      • KPPS2 antibody
      • PPKS 2 antibody
      • PPKS2 antibody
      see all

    Images

    • ab16434 staining A431 cells by ICC/IF. Cells were methanol fixed and blocked in 10% serum for 1 hour at 25°C. The primary antibody was diluted 1/100 (PBS+10%FBS) and incubated with the sample for 1 hour at 25°C. An Alexa Fluor® 488 conjugated goat anti-mouse antibody, diluted 1/1500, was used as the secondary.

      See Abreview

    • ab16434 staining Desmoplakin I+II in rat glial cultured cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde and blocked with 1% serum for 10 minutes at 25°C. Samples were incubated with primary antibody (1/200 in PBS + 5% serum) for 2 hours at 25°C. An Alexa Fluor® 488-conjugated donkey anti-mouse IgG polyclonal (1/300) was used as the secondary antibody.

      See Abreview

    • All lanes : Anti-Desmoplakin I+II antibody [DP2.15] (ab16434) at 1/500 dilution

      Lane 1 : Control MDA-MB-231 (human breast adenocarcinoma cell line) cell lysate
      Lane 2 : MDA-MB-231 (human breast adenocarcinoma cell line) treated with Desmoplakin I+II RNAi, cell lysate

      Lysates/proteins at 40 µg per lane.

      Developed using the ECL technique.

      Predicted band size: 331 kDa

      See Abreview

    References

    This product has been referenced in:
    • Clabault H  et al. Effects of selective serotonin-reuptake inhibitors (SSRIs) on human villous trophoblasts syncytialization. Toxicol Appl Pharmacol 349:8-20 (2018). WB . Read more (PubMed: 29679653) »
    • Kolahi K  et al. Real-Time Tracking of BODIPY-C12 Long-Chain Fatty Acid in Human Term Placenta Reveals Unique Lipid Dynamics in Cytotrophoblast Cells. PLoS One 11:e0153522 (2016). Read more (PubMed: 27124483) »
    See all 15 Publications for this product

    Customer reviews and Q&As

    1-10 of 10 Abreviews or Q&A

    Answer

    Thank you for your message and for providing this further information. Unfortunately, I am unsure which details about your protocol the mentioned publication explains. The expression of desmoplakin I+II in MDCK cells is of course not in question and studies of the various desmosomal components is absolutely feasible with the appropriate antibodies. This has nicely been demonstrated Mattey & Garrod (PMIDs 3793787 und 3793798) with polyclonal antibodies that had been generated in guinea-pig, which you certainly are aware of. As previously mentioned, our mouse monoclonal anti-Desmoplakin I+II antibody ab16434 has only been tested for its cross-reactivity with human, mouse, rat, cow and chicken. It has not been tested if it also recognises the target protein in MDCK cells, and we therefore cannot guarantee that positive signals can be obtained. You may want to test this in your experimental setting; however, in this case we unfortunately cannot provide a free of charge replacement or a credit note as the antibody is used in an untested species. Having said this, we would be pleased to arrange for a replacement or a credit note, if it turns out that the antibody is not working with positive controls such as human cells A432 and MCF-7. Also, we will happily offer technical support in an attempt to improve the results obtained, and therefore we would appreciate some more information on the sample type and preparation and which fixation methods have already been tried. Thank you for your cooperation. We appreciate your understanding of our policies. Please feel free to contact us again if you have any additional questions or concerns.

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    Application
    Immunocytochemistry
    Sample
    Rat Cultured Cells (Glial Cells)
    Specification
    Glial Cells
    Blocking step
    Serum as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 25°C
    Fixative
    Paraformaldehyde

    Abcam user community

    Verified customer

    Submitted Oct 06 2014

    Question
    Answer

    My colleague told us that she did meet you and that you explainedyour disappointment about ab16434: no staining in ICC/IF after adding glycerol as recommended.

    Iam sorry to hear you are experiencing difficulties with this product. We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.

    I am attaching our questionnaire so that we can gather further information regarding the samples tested and the protocol used. It will take you five to ten minutes to fill the questionnaire. Once we receive the completed questionnaire, we will look at the protocol and see if there are any suggestions we can make that may improve the results. This information will also allow us to investigate this case internally and initiate additional testing where necessary. If the product was purchased in the last six months and is being used according to our Abpromise, we would be happy to replace or refund the antibody.


    I look forward to receiving your reply.

    Read More

    Answer

    Thank you very much for your enquiry.

    Please find attached the standard protocols used with our antibodies. These protocols and further tips can also be found on our website (https://www.abcam.com/index.html?pageconfig=popular_protocols).

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

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    Answer

    Vielen Dank für Ihre Anfrage und diese weiteren Informationen. Leider bin ich mir nicht ganz sicher, welche Fragen die mitgeschickte Publikation beantwortet. Die Expression von Desmoplakin I+II in MDCK-Zellen steht außer Frage und eine Untersuchung der verschiedenen Desmosomenkomponenten ist mit geeigneten Antikörpern natürlich möglich. Dies wurde in den Publikationen von Mattey & Garrod (PMIDs 3793787 und 3793798) mit Antikörpern durchgeführt, die im Meerschwein generiert wurden. Leider wurde für unseren Maus-monoklonalen anti-Desmoplakin I+II- Antikörper ab16434 bislang nicht getestet, ob er auch die Zielproteine in MDCK-Zellen erkennt, und daher können wir leider nicht garantieren, dass man damit positive Signale erzielen kann. Der Endanwender kann dies gerne ausprobieren; leider können wir keinen Ersatz anbieten, sollte es nicht funktionieren, da der Antikörper in einer bisher nicht getesteten Anwendung oder Spezies verwendet wird. Allerdings sind wir gerne bereit, einen Ersatz oder eine Gutschrift zu arrangieren, falls sich anhand von Positivkontrollen wie zum Beispiel den humanen Zelllinien A432 and MCF-7 herausstellt, dass der Antikörper nicht funktioniert. Um Vorschläge zur Optimierung des Protokolls geben zu können, benötige ich weitere Informationen, zum Beispiel zur Probenvorbereitung oder bereits ausgetesteten Alternativen zur Fixierung. Vielen Dank für Ihre Kooperation. Bitte zögern Sie nicht, sich wieder bei uns zu melden, falls Sie weitere Fragen haben.

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    Answer

    Vielen Dank für Ihre Anfrage. Es tut mir leid, dass der Kunde Probleme mit unserem Anti-Desmoplakin I+II Antikörper ab16434 hatte. Leider ist ab16434 nicht für die Anwendung mit MDCK (dog cells) getestet und wir können nicht garantieren, dass er funktioniert. Wir sind jedoch sehr gerne bei der Optimierung des Protokolls behilflich, um eventuell eine Verbesserung der Ergebnisse zu erzielen. Daher hätte ich noch einige Fragen an den Kunden: I am sorry to hear you have had difficulty obtaining satisfactory results from this Desmoplakin I+II antibody. I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been successful. Unfortunately, this antibody has not been tested on MDCK cells yet and therefore we cannot guarantee that it works. However, we are pleased to offer technical support in an attempt to improve the results obtained. Thus, I would appreciate if you can confirm some further details: 1) What kind of difficulties do you encounter: no staining or non-specific staining? It would be great if you could provide an image which would help us to assess the results. 2) Could you provide some more details on the sample type and sample preparation, please? What kind of positive control did you run? 3) May I ask whether you tried other fixatives apart from 4% PFA, such as methanol or acetone? If it turns out that the antibody is not working under optimised conditions with the appropriate positive control (for example human cell line A432 or MCF7, as indicated on the datasheet), we will provide a credit note, free of charge replacement or refund, if the problem has been reported within 6 months of purchase. In order to proceed with this, could you please let me know the lot number, the order number or the date of purchase? Thank you for your cooperation. We look forward to your reply. Vielen Dank für Ihre Kooperation und Hilfsbereitschaft. Der Kunde kann natürlich auch gerne auf Deutsch oder Französisch antworten, falls das die Kommunikation erleichtert. Ich wünsche Ihnen ein schönes Wochenende!

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    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (A431)
    Specification
    A431
    Fixative
    Methanol
    Permeabilization
    No
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C

    Abcam user community

    Verified customer

    Submitted Oct 31 2008

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (MDA-MB-231 breast cancer cell line)
    Loading amount
    40 µg
    Specification
    MDA-MB-231 breast cancer cell line
    Gel Running Conditions
    Reduced Denaturing (gel 6 %)
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

    Abcam user community

    Verified customer

    Submitted Oct 08 2008

    Answer

    Thank you for your enquiry. I am sorry but we do not have any information where either of these antibodies have been applied in publications. Desmoplakin I+II antibody [2Q400] (ab16434) has been shown to work by Western blot, Immunohistochemistry (Frozen sections) and Immunocytochemistry. Desmoplakin antibody (ab14418) has been demonstrated to work by Western blot, Immunoprecipitation, ELISA and Immunofluorescence. We make an "Abpromise" to our customers such that we guarantee that our antibodies behave as we say they will. Should this not be the case we offer a replacement vial or credit note (provided the antibody has been purchased within the past 90 days). Unfortunately I do not have details as to the accession number that the cDNA used to generate the immunogen for ab14418 corresponds to. I can however tell you that ab14418 recognizes both Desmoplakins 1 and 2. These or any of our antibodies can be ordered by fax, e-mail (orders@abcam.com) or on line. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

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