Anti-Survivin DEx3 抗体 (ab3731)

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Immunocytochemistry/ Immunofluorescence - Anti-Survivin DEx3 antibody (ab3731)

    Key features and details

    • Rabbit polyclonal to Survivin DEx3
    • Suitable for: ICC/IF
    • Reacts with: Human
    • Isotype: IgG

    製品の概要

    • 製品名

      Anti-Survivin DEx3 antibody

    • 製品の詳細

      Rabbit polyclonal to Survivin DEx3

    • 由来種

      Rabbit

    • 特異性

      This antibody has been tested in Western blotting against equal loadings of purified wild type survivin, 2B and DEx3. When used at 1/250, it detects a single clean (but weak) band of the appropriate size for DEx3, but no band is detected with wild type survivin or 2B. We have not succeeded in detecting endogenous Survivin DEx3 using this antibody (Survivin DEx3 is expressed at very low levels). This antibody has been used successfully however, to detect overexpressed Survivin DEx3 in whole cell extracts.

    • アプリケーション

      適用あり: ICC/IFmore details

    • 種交差性

      交差種: Human

    • 免疫原

      Synthetic peptide corresponding to Human Survivin DEx3 aa 50-150.
      (Peptide available as ab13763)

    • ポジティブ･コントロール

      • This antibody gave a positive result in HeLa cell line in ICC/IF

    • 特記事項

      This antibody should be aliquoted to reduce freeze-thaw cycles as it loses performance quickly after multiple freeze-thaws.

      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.

    製品の特性

    • 製品の状態

      Liquid

    • 保存方法

      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.

    • バッファー

      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...

    • 精製度

      Immunogen affinity purified

    • ポリ/モノ

      ポリクローナル

    • アイソタイプ

      IgG

    • 研究分野

    アプリケーション

    Our Abpromise guarantee covers the use of ab3731 in the following tested applications.

    The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

    アプリケーション Abreviews 特記事項
    ICC/IF Use a concentration of 5 µg/ml.

    ターゲット情報

    • 関連性

      Survivin is expressed in G2/M phase in a cycle regulated manner. At the beginning of mitosis, survivin associates with mitotic spindle microtubules, this is essential for survivin's anti-apoptosis function. Survivin may counteract a default induction of apoptosis in G2 / M phase. Survivin over-expression in cancer may overcome this apoptotic checkpoint. Survivin DEx3 (lacking exon 3) and survivin-2B (retaining a part of intron 2 as a cryptic exon) are isoforms of survivin that behave differently to survivin. Skipping of exon 3 in survivin-Ex3 results in a modification of the BIR domain, followed by an additional frame shift in exon 4. In consequence, the open reading frame of Survivin DEx3 ends in the 3' untranslated region of survivin at position 581 and generates a novel 63 amino acids-long COOH-terminal tail in survivin-Ex3 coded by 87 nucleotides of exon 4 and 103 nucleotides of the original 3' untranslated region. See Mahotka et al. 1999 for more details.

    • 細胞内局在

      Cytoplasmic and Nuclear

    • 参照データベース

      • 別名

        • API4 antibody
        • Apoptosis Inhibitor 4 antibody
        • BIRC5 antibody
        • EPR1 antibody
        • IAP4 antibody
        • TIAP antibody
        see all

      画像

      • Immunocytochemistry/ Immunofluorescence - Anti-Survivin DEx3 antibody (ab3731)
        Immunocytochemistry/ Immunofluorescence - Anti-Survivin DEx3 antibody (ab3731)
        ICC/IF image of ab3731 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab3731, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

      参考文献 (6)

      ab3731 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

      ab3731 は 6 報の論文で使用されています。

      • Waligórska-Stachura J  et al. Survivin DEx3 as a biomarker of thyroid cancers: A study at the mRNA and protein level. Oncol Lett 13:2437-2441 (2017). IHC-P ; Human . PubMed: 28454416
      • Waligorska-Stachura J  et al. Decreased expression of survivin 2B in human pituitary adenomas. A preliminary study. Folia Histochem Cytobiol 55:21-25 (2017). PubMed: 28509313
      • Doucette T  et al. Survivin transcript variant 2 drives angiogenesis and malignant progression in proneural gliomas. Neuro Oncol N/A:N/A (2014). Mouse . PubMed: 24676140
      • Végran F  et al. Association of p53 gene alterations with the expression of antiapoptotic survivin splice variants in breast cancer. Oncogene 26:290-7 (2007). PubMed: 16847456
      • Noton EA  et al. Molecular analysis of survivin isoforms: evidence that alternatively spliced variants do not play a role in mitosis. J Biol Chem 281:1286-95 (2006). PubMed: 16291752
      • Caldas H  et al. Survivin splice variants regulate the balance between proliferation and cell death. Oncogene 24:1994-2007 (2005). PubMed: 15688031

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      1-4 of 4 Abreviews or Q&A

      Question

      Gerne bestätige ich Ihnen, dass ich die Antikörper für die IHC-P austesten möchte.

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      Answer

      DISCOUNT CODES: ab3729: xxxxx ab3731: xxxxx Ablaufdatum (jeweils): xxxx2012 Es freut mich zu hören, dass Sie unser Angebot unseren Antikörper ab3729 und ab3731 in der Immunhistochemie an Paraffinschnitten (IHC-P) zu testen, annehmen möchten. Wie am Telefon besprochen, können Sie den Gutscheincode jeweils zum Kauf eines weiteren Antikörpers einsetzen, sobald Sie uns ein Abreview für diesen Antikörper eingereicht haben. Die Discountcodes haben obige Nummern und können zum Kauf weiterer Produkte eingesetzt werden. Der Code wird jeweils gültig, sobald Sie uns ein Abreview für diesen Antikörper über den Test in der IHC-P eingereicht haben. Bitte geben Sie diesen Code auch im Abschnitt “Additional Notes” des Abreviews mit an, so dass wir wissen, dass sich dieses Abreview auf die Gutscheinaktion bezieht. Der Code wird dadurch aktiviert. Bitte kontaktieren Sie bei der nächsten Bestellung unsere Kundendienst mit dem Code und der ursprünglichen Bestellnummer. Bitte zögern Sie nicht, sich wieder an uns zu wenden, falls Sie weitere Fragen haben. Wir freuen uns auf Ihr Abreview, egal ob Ihre Ergebnisse positiv oder negativ sind und wünschen Ihnen viel Glück für Ihre Forschung. Die Bedingungen dieses Angebotes befinden sich unter dem folgenden Link: www.abcam.com/collaborationdiscount  

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      Question

      I am overexpressing human survivin delta Ex3 in a cell line.  Shouldn’t this antibody (ab76424) detect this form since it corresponds to the N terminus of survivin?  I have tried using the survivin DEx3 antibody (ab3731) but can’t detect it with that antibody either.  I appreciate any feedback you can give me.

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      Answer

      Thank you for contacting Abcam regarding ab76424 and ab3731. I am sorry that you have been experiencing difficulties getting these antibodies to work in WB.  I am following up with the lab regarding reactivity of ab76424 with Delta Ex3.  Regarding ab3731, one point to note is that this antibody only works well in Western blotting on cell lysates overexpressing DEx3(see PMID 16847456), and it first requires IP with another antibody, before blotting with this antibody. It has been previously used with an IP performed from 5 million cells overexpressing DEx3.  If you have not performed an IP, the antibody will not likely detect the protein in WB. I hope this information is helpful.  Please do not hesitate to contact us if you have any additional questions.

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      Question

      I am getting a band ~35 kDa. The blot is clean without any background and that is the only band I am getting. Please let me know whether this antibody has some non-specific binding around 35 kDa. I dont think the signals I am getting at 35 kDa are non-specific (Because the intensity of the bands are not same - I have loaded equal amount of protein - based on actin and BCA assay).

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      Answer

      Thank you for your email. We have not had any reports of a band being detected at approximately 35 kDa with ab3731. We do sell the blocking peptide for this antibody which should be helpful for you - it is ab13763. Interestingly, it is noted for ab469 - Rabbit polyclonal to Survivin - that in addition to the survivin band, additional bands are seen at approximately 37 and 50 kDa. However, we do not know what these bands are. Please note that we have not succeeded in detecting endogenous Survivin DEx3 using ab3731 (Survivin DEx3 is expressed at very low levels). This antibody has been used successfully however, to detect overexpressed Survivin DEx3 in whole cell extracts. This antibody only works well in Western blotting on cell lysates overexpressing DEx3, and it first requires IP with another antibody, before blotting with this antibody. It has been previously used with an IP performed from 5 million cells overexpressing DEx3. I hope this helps; if you have any additional questions, please contact us again.

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      Question

      What size band is detected with this antibody in WB?

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      Answer

      Thank you for your enquiry. Ab3731 should detect a band of ~ 14 kDa. Please let me know if you have any additional questions or require additional assistance.

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