Overview

  • Product name

  • Description

    Rabbit polyclonal to DEK
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IP, WBmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    9aa synthetic peptide from the region between aa 178-198 of Human DEK

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Storage buffer

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituent: 0.87% Sodium chloride

    Without Mg2+ and Ca2+
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab74975 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
IP Use at an assay dependent dilution.
WB 1/500. Predicted molecular weight: 43 kDa.

Target

  • Function

    Involved in chromatin organization.
  • Tissue specificity

    Ubiquitous. Expressed at relatively high levels.
  • Involvement in disease

    A chromosomal aberration involving DEK is found in a subset of acute myeloid leukemia (AML); also known as acute non-lymphocytic leukemia (PubMed:1549122). Translocation t(6;9)(p23;q34) with NUP214/CAN (PubMed:1549122). It results in the formation of a DEK-NUP214 fusion gene (PubMed:1549122).
  • Sequence similarities

    Contains 1 SAP domain.
  • Post-translational
    modifications

    Phosphorylated by CK2. Phosphorylation fluctuates during the cell cycle with a moderate peak during G(1) phase, and weakens the binding of DEK to DNA.
  • Cellular localization

    Nucleus. Enriched in regions where chromatin is decondensed or sparse in the interphase nuclei.
  • Information by UniProt
  • Database links

  • Alternative names

    • D6S231E antibody
    • Dek antibody
    • DEK gene antibody
    • DEK oncogene antibody
    • DEK oncogene DNA binding antibody
    • DEK_HUMAN antibody
    • Protein DEK antibody
    see all

References

This product has been referenced in:

  • Sandén C  et al. The DEK oncoprotein binds to highly and ubiquitously expressed genes with a dual role in their transcriptional regulation. Mol Cancer 13:215 (2014). CHIPseq ; Human . Read more (PubMed: 25216995) »
See 1 Publication for this product

Customer reviews and Q&As

1-9 of 9 Abreviews or Q&A

Answer

According to our records, ab74975 was proving difficult to use in Western blot and we were in contact in order to help resolve the issue.
Looking at our correspondence, it appears that we are awaiting more details in order to help us better understand the difficulties experienced. If the requested information has already been sent, it appears that it did not reach our Scientific Support team and we apologize for this inconvenience. In this case we would like to ask for the information again so that we can reach a resolution.

If the issue has already been settled, please let us know so that we can be assured that the problem has been solved to your satisfaction and update our records.

We wish you the best of luck with your research and look forward to a reply.

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Answer

Thank you for contacting us.
I am sorry to hear you are experiencing difficulties with one of our products. We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.
We had so far only one other negative feedback in Western blot (the one in question 4). In order to be able to see whether the vial might be faulty, I would need to investigate firstly the results in more details. It is also difficult for me to make recommendations, without seeing the whole protocole. Also, I would appreciate to know more about the sample treatment. Please do also note what the batch number of your vial is, whether it is the same as the one from the previous complaint.
I am attaching therefore our questionnaire so that we can gather further information regarding the samples tested and the protocol used. 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.

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Abreviews
Application
ChIP
Sample
Human Cell lysate - nuclear (Leukemia cell line)
Specification
Leukemia cell line
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 15 minute(s) and 0 second(s)
Specification of the cross-linking agent: 1% formaldehyde
Detection step
Other
Positive control
Nuclear lysate of untreated U937 cells.

Abcam user community

Verified customer

Submitted Jan 21 2012

Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (testis)
Specification
testis
Fixative
Formaldehyde
Permeabilization
Yes - Triton X-100 0,5%
Blocking step
BSA as blocking agent for 45 minute(s) · Concentration: 3% · Temperature: 22°C

Abcam user community

Verified customer

Submitted Jan 16 2012

Answer

Thank you very much for your interest in ab74975. To our knowledge, ab74975 has not been tested in ChIP. I can also offer a discount off a future purchase if you buy ab74975 now, test it in ChIP and submit 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 feedback. The discount would be to the value of 1 free primary antibody. If you are interested in this offer, please follow these steps: 1. Reply to this e-mail to let me know that you would like to proceed and test ab74975 in ChIP. I will then send a discount code. This code must be issued before purchasing ab74975 so please wait for my reply before ordering. 2. Purchase ab74975 either by phone, fax, or online (www.abcam.com). 3. Test it in ChIP. 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 new order by phone, fax, or on the web and mention the discount code. The discount can be redeemed for any primary antibody 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 ab74975 turns out to be unsuitable for ChIP, you will still receive the discount on your next purchase after your Abreview has been submitted. Please let me know if you 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.

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Answer

Merci pour ces précisions. En effet, l'anti-DEK ab83099 est également un bon alternatif. Le numéro de commande de remplacement gratuit (une unité de ab83099) est : *******. Vous recevrez prochainement un mail de confirmation comprenant les détails d'expédition. N'hésitez pas à nous contacter lors d'une prochaine occasion.  

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Answer

Je parle effectivement français. Je suis désolé d'apprendre que les 2 anti-DEK ab22826 et ab76975 que vous avez testés ne fonctionnent pas comme indiqué sur les fiches techniques respectives. Votre protocole me semble bon, je crains que vous ayez reçu 2 fois des anticorps défectueux. Désirez-vous recevoir un autre anticorps anti-DEK, tel que ab105175 (www.abcam.com/ab105175) ou un remboursement de la commande originale?  Merci de m'indiquer la solution qui vous conviendrait le mieux.  

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Question

LOT NUMBER GR26610-6 ORDER NUMBER FOCR CA-213621 DESCRIPTION OF THE PROBLEM Multiple bands. I used 2 nuclear extracts (mouse testicular nuclear extract and human fibroblast nuclear extract)and loaded both in two wells each. I transfered the proteins on a itrocellulose membrane at 100V for 1h at 4°C. I cut my membrane in half (each half with a ladder and both nuclear extracts). I performed western blot on both using ab22826 on one half and ab74975 on the other. After secondary antibody (anti-rabbit 800) revealed on a Li-cor apparatus, I don't find any principal band at 42-43 KDa on any membrane, neither human or mouse. The banding pattern on each membrane (exactly the same nuclear extracts) is completely different. SAMPLE 1- mouse testicular nuclear extract (directly removed from the animal) 2- human primary fibroblasts nuclear extract (from a cell culture) PRIMARY ANTIBODY 1st MEMBRANE: ab74975 (Rabbit polyclonal to DEK) from abcam, dilution 1/500 in 5% milk in 1x PBS, incubation O/N at 4°C under rotation. Rinsed once in 1x PBS + 3x5 minutes in 1x PBS DETECTION METHOD fluorescence detection methods with The Odyssey® Infrared Imaging System POSITIVE AND NEGATIVE CONTROLS USED This protein is supposed to be ubiquitous and present in any nucleous. I used human fibroblasts as seen in many publications. ANTIBODY STORAGE CONDITIONS Antibody placed at -20°C at reception, no multiple freeze and thaw SAMPLE PREPARATION 2x SDS page sample buffer: Mix 10 ml 1.5 M Tris (pH 6.8), 6 ml 20% SDS, 30 ml glycerol,15 ml β-mercaptoethanol and 1.8 mg bromophenol blue. Adjust volume to 100 ml with H2O. Aliquot in 10 ml stock solution and store at – 20°C. Store working solution at 4°C. I diluted my samples 1/2 in the 2x SDS page sample buffer and boiled them for 10 minutes before loading. In the nuclear extracts, there is a cocktail of proteases inhibitors ("COmplete" from Roche). AMOUNT OF PROTEIN LOADED 50µg for each nuclear extract in each well ELECTROPHORESIS/GEL CONDITIONS 12% acrylamide gel from this recipe: H2O 1,6mL 30% acryl-bisacrylamide mix 2,0mL 1.5 M Tris (pH 8.8)1,3mL 10% SDS 0,05mL 10% ammonium persulfate 0,05mL TEMED 0,002mL Running buffer: 1x solution from this 10x solution: 10 x SDS PAGE Running buffer Dissolve 10 g SDS, 30.3 g Tris and 144.1 g glycin in 800 ml H2O. Adjust volume to 1 liter with H2O. Store at room temperature. TRANSFER AND BLOCKING CONDITIONS 1x Transfer Buffer Dissolve 2.9 g Glycine, 5.8 g Tris and 0.37 g SDS in 200 ml methanol. Adjust volume to 1 liter with H2O. Store at 4°C. trandfered 1h at 100V at 4°C on a nitrocellulose membrane Blocking: 5% powdred skim milk in 1x PBS for 1h at RT SECONDARY ANTIBODY IRdye 800CW goat anti-rabbit IgG (H+L), LiCor, 1/10000 in 5% milk in 1x PBS for 1h at RT under agitation in the dark. (manufacturer protocol was followed for the reconstitution and storage of this antibody. Always work well) HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 2 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? increase the µg of protein loaded ADDITIONAL NOTES Remember that I used exactly the same extracts on both membranes. I boiled each extract with the SDS sample buffer and splitted each tubes in two different wells on the same gel. The transfer was performed before cutting the membrane or the gel. Only the 1st incubation (primary antibody) and the wash after this primary antibody were separated, due to the different primary antibodies. This protein is known to di- and multi-merize. Do you think that maybe it could resist the denaturing and reducing agents in the buffers and that the band at ~80KDa could be a dimer? I didn't overexpose my membranes. Signals were really sharp and strong. The website tells that you tested both antibodies in western blots. What samples were you using?

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Answer

Thank you for taking time to complete our questionnaire and for contacting us. The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality. I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been successful. Having reviewed the protocol details, I believe this product should have given satisfactory results. It appears that you may have received a faulty vial. Could you please confirm that only ab74975 did not work and that you obtained a correct band with ab22826? I apologize for the inconvenience and am pleased to offer you a free of charge replacement, credit note, or refund in compensation. Thank you for your cooperation. I look forward to hearing from you with details of how you would like to proceed.

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Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (knees - sections prepared from the whole knee)
Specification
knees - sections prepared from the whole knee
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.05% Triton X-100
Blocking step
Serum as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Jul 15 2011

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