Anti-RYBP antibody (ab5976)

Reviews (3)Q&A (4)References (7)

Overview

  • Product name
    Anti-RYBP antibody
    See all RYBP primary antibodies
  • Description
    Rabbit polyclonal to RYBP
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IHC-P, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide:

    TPKGDMSAVNDESF

    , corresponding to amino acids 215-228 of Human DEDAF.

    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI
  • Positive control
    • A549, HepG2 or 3T3 whole cell lysate.
  • General notes
    Stable for one year.


    Apoptosis is related to many diseases and induced by a family of cell death receptors and their ligands. Cell death signals are transduced by death domain (DD) death effector domain (DED), and caspase recruitment domain (CARD) containing molecules. Several molecules including caspases and adaptor FADD contain DEDs. A novel protein that interacts with DED of caspase 8 and 10, and FADD was identified recently and designated DEDAF for DED associated factor. DEDAF is identical to the transcriptional repressor RYBP. DEDAF/RYBP is expressed in multiple tissues and cell lines. DEDAF interacts with FADD and augments the formation of CD95/FADD/capase 8 complexes at the cell membrane, and interacts with DED-containing DNA biding protein (DEDD) in the nucleus indicating it is involved in the regulation of both cytoplasmic and nuclear events of apoptosis.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C.
  • Storage buffer
    Preservative: 0.02% Sodium azide
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Purification notes
    DEDAF Antibody is affinity chromatography purified via peptide column.
  • Primary antibody notes
    Apoptosis is related to many diseases and induced by a family of cell death receptors and their ligands. Cell death signals are transduced by death domain (DD) death effector domain (DED), and caspase recruitment domain (CARD) containing molecules. Several molecules including caspases and adaptor FADD contain DEDs. A novel protein that interacts with DED of caspase 8 and 10, and FADD was identified recently and designated DEDAF for DED associated factor. DEDAF is identical to the transcriptional repressor RYBP. DEDAF/RYBP is expressed in multiple tissues and cell lines. DEDAF interacts with FADD and augments the formation of CD95/FADD/capase 8 complexes at the cell membrane, and interacts with DED-containing DNA biding protein (DEDD) in the nucleus indicating it is involved in the regulation of both cytoplasmic and nuclear events of apoptosis.
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab5976 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 Use a concentration of 0.5 - 1 µg/ml. Detects a band of approximately 32 kDa.Can be blocked with RYBP peptide (ab6097). Can be blocked with DEDAF peptide (human) .
IHC-P Use a concentration of 5 - 10 µg/ml.
ICC/IF Use a concentration of 1 - 20 µg/ml.

Target

  • Function
    May be implicated in the regulation of the transcription as a repressor of the transcriptional activity of E4TF1. In tumor cell lines, may induce apoptosis.
  • Tissue specificity
    Widely expressed with highest levels in lymphoid tissues and placenta.
  • Sequence similarities
    Contains 1 RanBP2-type zinc finger.
  • Cellular localization
    Nucleus. Cytoplasm. Primarily found in the nucleus.
  • Information by UniProt
  • Database links
    see all
  • Alternative names
    • 2410018J24Rik antibody
    • AAP 1 antibody
    • AAP1 antibody
    • APAP 1 antibody
    • APAP-1 antibody
    • APAP1 antibody
    • Apoptin associating protein 1 antibody
    • Apoptin-associating protein 1 antibody
    • cb337 antibody
    • Death effector domain associated factor antibody
    • Death effector domain-associated factor antibody
    • Death effector domain-associated factor B antibody
    • DED associated factor antibody
    • DED-associated factor antibody
    • DED-associated factor B antibody
    • DEDAF antibody
    • MGC62213 antibody
    • RING1 and YY1 binding protein a antibody
    • RING1 and YY1 binding protein antibody
    • RING1 and YY1-binding protein antibody
    • RING1 and YY1-binding protein B antibody
    • Ring1 interactor RYBP antibody
    • RYBP antibody
    • RYBP_HUMAN antibody
    • rybpa antibody
    • wu:fi34f03 antibody
    • YEAF 1 antibody
    • YEAF1 antibody
    • YY1 and E4TF1 associated factor 1 antibody
    • YY1 and E4TF1-associated factor 1 antibody
    • YY1- and E4TF1/GABP-associated factor 1 antibody
    • zgc:153842 antibody
    see all

Images

  • Western blot - Anti-RYBP antibody (ab5976)
    Western blot - Anti-RYBP antibody (ab5976)

    Western blot analysis of DEDAF expression in human A549 (lane A), HepG2 (lane B), and mouse 3T3 (lane C) cell lysates withab5976 at 1 µg /ml.

    Western blot analysis of DEDAF expression in human A549 (lane A), HepG2 (lane B), and mouse 3T3 (lane C) cell lysates withab5976 at 1 µg /ml.
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RYBP antibody (ab5976)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RYBP antibody (ab5976)

    Immunohistochemistry of DEDAF in mouse liver tissue with DEDAF antibody at 5 µg/ml.

  • Immunocytochemistry/ Immunofluorescence - Anti-RYBP antibody (ab5976)
    Immunocytochemistry/ Immunofluorescence - Anti-RYBP antibody (ab5976)This image is courtesy of an Abreview submitted by Dr. Ligia Tavares
    ab5976 staining RYBP in Mouse embryonic stem cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with Triton 0.5% and blocked with 1% milk for 30 minutes at 22°C. Samples were incubated with primary antibody (1/1000) for 1 hour at 22°C. An Alexa Fluor®488-conjugated Goat anti-rabbit IgG (H+L) polyclonal (1/400) was used as the secondary antibody.
  • Immunocytochemistry/ Immunofluorescence - Anti-RYBP antibody (ab5976)
    Immunocytochemistry/ Immunofluorescence - Anti-RYBP antibody (ab5976)
    Immunofluorescence of DEDAF in A549 cells using ab5976 at 20 ug/ml.

References

This product has been referenced in:
  • Bian F  et al. Polycomb repressive complex 1 controls uterine decidualization. Sci Rep 6:26061 (2016). Read more (PubMed: 27181215) »
  • Eid A & Torres-Padilla ME Characterization of non-canonical Polycomb Repressive Complex 1 subunits during early mouse embryogenesis. Epigenetics 11:389-97 (2016). Read more (PubMed: 27081692) »
See all 7 Publications for this product

Publishing research using ab5976? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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1-7 of 7 Abreviews or Q&A

Question

Worked with it before, one vial was brilliant, two showed unspecific signal.
Tried than ab89603 Anti-RYBP antibody, no success either. Tried many optimazation steps for WB and IHC.
human samples, lynphocites and tonsils,

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Answer

Ich habe Rückmeldung aus dem Labor erhalten: Leider sieht es so aus, dass der ab128104 Anti-RYBP antibody [1B2] nur gegen das rekombinante Protein getestet wurde.

Soll ich Ihnen daher (wie besprochen) den ab107896 Anti-RYBP antibody zukommen lassen?

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Immunocytochemistry/ Immunofluorescence abreview for Anti-RYBP antibody

 Good
Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (mouse embryonic stem cells)
Specification
mouse embryonic stem cells
Fixative
Formaldehyde
Permeabilization
Yes - 0.5 % triton
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 22°C
Read More

Dr. Ligia Tavares

Verified customer

Submitted Sep 16 2011

ChIP abreview for Anti-RYBP antibody

 Below Average
Abreviews
Abcam has not validated the combination of species/application used in this Abreview.
abreview image
Application
ChIP
Sample
Mouse Cell lysate - whole cell (ES cells)
Specification
ES cells
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: formaldehyde
Detection step
Real-time PCR
Read More

Abcam user community

Verified customer

Submitted Mar 31 2011

Western blot abreview for Anti-RYBP antibody

 Good
Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
abreview image
Application
Western blot
Sample
Mouse Cell lysate - nuclear (ES cells)
Loading amount
100000 cells
Specification
ES cells
Gel Running Conditions
Reduced Denaturing (12%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Read More

Dr. Ligia Tavares

Verified customer

Submitted Mar 18 2011

Question

here is another try at sending the western with DEDAF. The attachment is a picture of the western blot using the DEDAF antibody. Used at a concentration of 0.5 ug/ml in IMR90 and HEP-G2 cells.

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Answer

Thank you again for sending the image of your blot. I contacted the originator of this antibody, and they have never seen that additional band at 60 kDa. You do seem to be detecting DEDAF as you are seeing a band at approx 32 kDa and most likely the band at 60 kDa is non-specific or perhaps a splicing variant or cross-reaction to another family member. The replacement vial is being shipped on order# 80151 and it will take approximately 1 week for you to receive it as it is currently out of stock. Please let me know how the replacement works out for you.

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Question

BATCH NUMBER 68529 ORDER NUMBER 05344 DESCRIPTION OF THE PROBLEM Multiple bands SAMPLE MCF-10As, MCF-7s, T47Ds nuclear extracts IMR-90, HepG2 whole cell lysates PRIMARY ANTIBODY Abcam,Dedaf,PBS-T w/ 1% BSA,0.7ug/ml,overnight(4 degrees celecius) wash:4xPBS-T, 30 mins each SECONDARY ANTIBODY Sigma, Goat- anti-rabbit IgG horse radish peroxidase(GAR-HRP),PBS-T w/ 1% BSA,1:10000,1h (room temp) wash:4xPBS-T, 30 mins each DETECTION METHOD ECL POSITIVE AND NEGATIVE CONTROLS USED HepG2 whole cell lysate as positive control no primary control ANTIBODY STORAGE CONDITIONS 4 degrees celcius SAMPLE PREPARATION laemmli buffer heated at 90 degrees celcius for 3 mins extracts prepared with protease inhibitors: Trasylol, Leu/Pep, DTT, PMSF, Sodium Fluoride, Sodium Orthovanadate, beta-Glycerophosphate AMOUNT OF PROTEIN LOADED 10-20 micrograms ELECTROPHORESIS/GEL CONDITIONS 10% acrylamide gel TRANSFER AND BLOCKING CONDITIONS transer: semi-dry, room temp, 20 volts, 0.4 amp limit, 2 hours block: 1% BSA at room temperature for 1 hour HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 4 HAVE YOU RUN A "NO PRIMARY" CONTROL? Yes DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? more washes after primary and secondary antibody incubations ADDITIONAL NOTES one band at around 32 kd (assumed to be DEDAF) and a second just under the 62 kd marker

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Answer

Thank you for the details that you have submitted. From the description, it seems that you are detecting DEDAF at 32 kDa and are also seeing a non-specific band. I would suggest that you try decreasing the antibody concentration and/or the incubation period for both the primary and secondary antibodies. What were the results with the secondary-only control? Also, we do sell the blocking peptide for this antibody - use the blocking peptide to differentiate between specific and non-specific bands. Only specific bands should be blocked (and thus disappear). If you have any additional questions, please contact us again.

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Question

Customer is using this antibody in Western blotting with 4 different breast cancer cell lines (MCF7 and MCF10A among them). He is seeing the expected band at 32 kDa, but is also seeing a band at approximately 60 kDa. Has this been seen before?

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Answer

Thank you for your phone call. I contacted the originator of this antibody and they were able to provide me with the following information. First, they said that with the A549, HepG2 and 3T3 whole cell lysates, they only observed a band at 32 kDa (there were no additional bands). Second, they have not tested this antibody in breast cancer cell lines, however, there may be a larger band in MOLT4 cells, but it is too faint to tell. I hope this is useful, if you have any additional questions please do let us know.

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