Anti-Survivin DEx3 antibody (ab3731)

Submit a review Q&A (4)References (5)

Overview

  • Product name
    Anti-Survivin DEx3 antibody
  • Description
    Rabbit polyclonal to Survivin DEx3
  • Host species
    Rabbit
  • Specificity
    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.
  • Tested applications
    Suitable for: WB, ICC/IFmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide of Human Survivin DEx3, derived from amino acids 50-150.

    Read Abcam's proprietary immunogen policy (Peptide available as ab13763.)

  • Positive control
    • This antibody gave a positive result in HeLa cell line in ICC/IF
  • General notes
    This antibody should be aliquoted to reduce freeze-thaw cycles as it loses performance quickly after multiple freeze-thaws.

Properties

  • Form
    Liquid
  • Storage instructions
    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.
  • Storage buffer
    Preservative: 0.02% Sodium Azide
    Constituents: 1% BSA, PBS, pH 7.4
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

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.

Application Abreviews Notes
WB 1/1000 - 1/5000. Predicted molecular weight: 17 kDa.
ICC/IF Use a concentration of 5 µg/ml.

Target

  • Relevance
    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.
  • Cellular localization
    Cytoplasmic and Nuclear
  • Database links
    • Alternative names
      • API4 antibody
      • Apoptosis Inhibitor 4 antibody
      • BIRC5 antibody
      • EPR1 antibody
      • IAP4 antibody
      • TIAP antibody
      see all

    Images

    • 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.

    References

    This product has been referenced in:
    • 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 . Read more (PubMed: 28454416) »
    • Doucette T  et al. Survivin transcript variant 2 drives angiogenesis and malignant progression in proneural gliomas. Neuro Oncol N/A:N/A (2014). Mouse . Read more (PubMed: 24676140) »
    See all 5 Publications for this product

    Publishing research using ab3731? Please let us know so that we can cite the reference in this datasheet.

    Customer reviews and Q&As

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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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