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RabMAb

Anti-Dishevelled 3 antibody [EP1991Y] (ab76081)

Reviews (2)Q&A (2)References (6)
Western blot - Anti-Dishevelled 3 antibody [EP1991Y] (ab76081)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dishevelled 3 antibody [EP1991Y] (ab76081)
  • Immunocytochemistry/ Immunofluorescence - Anti-Dishevelled 3 antibody [EP1991Y] (ab76081)
  • Flow Cytometry (Intracellular) - Anti-Dishevelled 3 antibody [EP1991Y] (ab76081)

Key features and details

  • Rabbit monoclonal [EP1991Y] to Dishevelled 3
  • Suitable for: WB, IHC-P, ICC/IF, Flow Cyt (Intra)
  • Reacts with: Rat, Human
  • Isotype: IgG

Conjugates logo Related conjugates and formulations

Carrier Free

Overview

  • Product name

    Anti-Dishevelled 3 antibody [EP1991Y]
    See all Dishevelled 3 primary antibodies

  • Description

    Rabbit monoclonal [EP1991Y] to Dishevelled 3

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IHC-P, ICC/IF, Flow Cyt (Intra)more details
    Unsuitable for: IP

  • Species reactivity

    Reacts with: Rat, Human
    Predicted to work with: Mouse

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: K562 cell lysate IHC-P: Human brain tissue ICC/IF: PC12 cells Flow Cyt: HeLa cells

  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. 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, along with publications, customer reviews and Q&As

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab76081 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)
1/500 - 1/1000. Predicted molecular weight: 78 kDa.
IHC-P (1)
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF
Use a concentration of 5 µg/ml.
Flow Cyt (Intra)
1/30.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
Notes
WB
1/500 - 1/1000. Predicted molecular weight: 78 kDa.
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF
Use a concentration of 5 µg/ml.
Flow Cyt (Intra)
1/30.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
Application notes
Is unsuitable for IP.

Target

  • Function

    May play a role in the signal transduction pathway mediated by multiple Wnt genes.

  • Sequence similarities

    Belongs to the DSH family.
    Contains 1 DEP domain.
    Contains 1 DIX domain.
    Contains 1 PDZ (DHR) domain.

  • Post-translational
    modifications

    Ubiquitinated. Deubiquitinated by CYLD, which acts on 'Lys-63'-linked ubiquitin chains.

  • Cellular localization

    Cytoplasm.
  • Information by UniProt

  • Database links

    see all

  • Alternative names

    • Dishevelled 3 (homologous to Drosophila dsh) antibody
    • Dishevelled 3 antibody
    • Dishevelled 3, dsh homolog (Drosophila) antibody
    • Dishevelled, dsh homolog 3 (Drosophila) antibody
    • Dishevelled-3 antibody
    • DSH homolog 3 antibody
    • dvl3 antibody
    • DVL3_HUMAN antibody
    • KIAA0208 antibody
    • OTTHUMP00000210777 antibody
    • OTTHUMP00000210778 antibody
    • Segment polarity protein dishevelled homolog DVL 3 antibody
    • Segment polarity protein dishevelled homolog DVL-3 antibody
    see all

Images

  • Western blot - Anti-Dishevelled 3 antibody [EP1991Y] (ab76081)
    Western blot - Anti-Dishevelled 3 antibody [EP1991Y] (ab76081)
    Anti-Dishevelled 3 antibody [EP1991Y] (ab76081) at 1/1000 dilution + K562 cell lysate at 10 µg

    Secondary
    Goat anti-Rabbit HRP labelled IgG at 1/2000 dilution

    Predicted band size: 78 kDa
    Observed band size: 95 kDa why is the actual band size different from the predicted?

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dishevelled 3 antibody [EP1991Y] (ab76081)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dishevelled 3 antibody [EP1991Y] (ab76081)

    Immunohistochemical staining of paraffin-embedded human brain tissue, using ab76081, at 1/100-1/250 dilution.

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Immunocytochemistry/ Immunofluorescence - Anti-Dishevelled 3 antibody [EP1991Y] (ab76081)
    Immunocytochemistry/ Immunofluorescence - Anti-Dishevelled 3 antibody [EP1991Y] (ab76081)

    ICC/IF image of ab76081 stained PC12 cells. The cells were 100% methanol fixed (5 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 (ab76081, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 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.

  • Flow Cytometry (Intracellular) - Anti-Dishevelled 3 antibody [EP1991Y] (ab76081)
    Flow Cytometry (Intracellular) - Anti-Dishevelled 3 antibody [EP1991Y] (ab76081)

    Overlay histogram showing HeLa cells stained with ab76081 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab76081, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

     

References (6)

Publishing research using ab76081? Please let us know so that we can cite the reference in this datasheet.

ab76081 has been referenced in 6 publications.

  • Zeng X  et al. Secreted Frizzled Related Protein 2 Modulates Epithelial-Mesenchymal Transition and Stemness via Wnt/ß-Catenin Signaling in Choriocarcinoma. Cell Physiol Biochem 50:1815-1831 (2018). PubMed: 30396168
  • Hua Y  et al. Oligomerization of Frizzled and LRP5/6 protein initiates intracellular signaling for the canonical WNT/ß-catenin pathway. J Biol Chem 293:19710-19724 (2018). PubMed: 30361437
  • Wu Y  et al. Low expression of secreted frizzled-related protein 2 and nuclear accumulation of ß-catenin in aggressive nonfunctioning pituitary adenoma. Oncol Lett 12:199-206 (2016). WB ; Human . PubMed: 27347125
  • He H  et al. miR-204-5p promotes the adipogenic differentiation of human adipose-derived mesenchymal stem cells by modulating DVL3 expression and suppressing Wnt/ß-catenin signaling. Int J Mol Med 35:1587-95 (2015). WB . PubMed: 25847080
  • Lin X  et al. Differential Expression of Wnt Pathway Genes in Sporadic Hepatocellular Carcinomas Infected With Hepatitis B Virus Identified With OligoGE Arrays. Hepat Mon 13:e6192 (2013). PubMed: 23483081
  • Burcklé C  et al. Control of the Wnt pathways by nephrocystin-4 is required for morphogenesis of the zebrafish pronephros. Hum Mol Genet 20:2611-27 (2011). IHC (PFA fixed) . PubMed: 21498478

Customer reviews and Q&As

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1-4 of 4 Abreviews or Q&A

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Dishevelled 3 antibody [EP1991Y]

 Average
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Cow Tissue sections (oviductal tissue)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris-EDTA, pH 9.0
Permeabilization
No
Specification
oviductal tissue
Blocking step
Serum as blocking agent for 20 minute(s) · Concentration: 2.5% · Temperature: 25°C
Fixative
Paraformaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Sep 28 2016

Western blot

 Good
Abreviews
abreview image
Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing (12 % SDS PAGE gel)
Sample
Human Cell lysate - whole cell (HEK 293T cells)
Specification
HEK 293T cells
Treatment
transfected with Dvl-3 plasmid
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted May 23 2013

Question

bonjour
je vous mets en pièce jointe l’article publié avec cet anticorps nous n’avons plu ce marquage au basal body mais un marquage cytroplasmique
alors que nous l’avions de manière récurrente depuis l’article.
merci de votre attention
cordialement
‘



voici le questionnaire rempli



Ce questionnaire ne prend que 5 minutes à être rempli. Pourriez-vous
remplir chaque champ afin que nous puissions mieux vous renseigner

1) Code du produit Abcam
Ab76081

2) Numéro de commande Abcam ou le numéro de lot/batch lot Gr41734

2) Description du problème
3) Marquage cytoplasmique et non basal body voir image jointe

4 Préparation de l’échantillon : Nh4cl 15mn blocage te hybridation
dans pbs, bsa3%
Espèce chien
Type d’échantillon :, culture cellulaire,:
Préparation de l’échantillon fixation PFA 4%,
Contrôle positif
Contrôle négatif secondaire seul

5) Etape de fixation : Oui
Si oui : agent fixateur et concentration PFA 4%,

Temps de fixation 20mn
Température de fixation 20°C

6) Méthode pour l’antigen retrieval

7) Etape de perméabilisation :
Avez-vous procéder à une étape de permabilisation (à détailler) ou
avez-vous additionné un agent de perméabilisation dans plusieurs
tampons de dilution ?
Agent de perméabilisation et concentration

8) Agent bloquant (ex BSA, serum…): NH4cl 15mn blocage et hybridation
dans PBS, bsa 3%
Concentration
Temps de saturation 1h
Température de saturation

9) Blocage des peroxidases endogènes ?
Blocage des biotines endogènes ?

10) Anticorps primaire (utilisation de plusieurs anticorps,
description dans “notes supplémentaires”) :
Concentration et dilution 1/50
Tampon de dilution o/N 4°C
Temps d’incubation

11) Anticorps secondaire :alexa fluor 555 molecular probes
Espèce donkey
Réagit contre rabbit
Concentration ou dilution 1/200
Tampon de dilution 1h
Temps d’incubation
Température d’incubation room température
Fluorochrome or enzyme conjuguée

12) Lavage après les anticorps primaire et secondaire :
Tampon pbs
Nombre de lavage 3

13) Méthode de détection

14) Combien de fois avez vous essayer cette application ?
Acs utilisé sans problème déja publié marque le basal body voir panel
B de la figure
Deuxième article en cours de révision toujours le même marquage sans
soucis sauf avec ce lot, alors que nous avons bien ce marquage avec
l’aC anti dvl2 (biol mol)
Obtenez-vous toujours le même résultat ?
Quelle étape avez vous optimisée ?

Read More
Answer

Merci d'avoir pris le temps de remplir notre questionnaire. Je suis désolée que vous avez eu des difficultés à obtenir de bons résultats avec ce nouveau lot.

Malheureusement nous ne trouvons pas le lot GR41734 - nous pensons que ceci doit probablement être le ab14173-4. Ma collègue vient aussi d'appeler votre service de commande pour vérifier ceci. A la date du 26 Janvier nous avons une commande chez vous du ab76081 lot GR14173-4. Nous n'avons malheureusement pas une autre production en stock.

Nous avons pas eu des autres retours négatives sur ce lot pendant les deux dernières années et il se peux donc que le problème soit du au tube que vous avez reçu et non au lot. Est-ce que vous voudriez essayer un nouveau tube du même lot? J'ai demandé au laboratoire si un nouveau lot est en production et je vous tiens au courant dès que j'ai des nouvelles.
J'ai aussi trouvé une commande d'un tube du lot GR14173-3 de vous en octobre dernier - ce tube avait bien fonctionné?

Effectivement, nos numéros de lot sont composés du lot de production (GR14173) ainsi que de la date de aliquotage/stockage (-3, ou -4). Le lot que nous avons en stock actuellement est le GR14173-5 et GR14173-6.

Est-ce que je peux vous poser quelques questions techniques additionnelles pour nous?
1.) Est-ce que vous avez changé des autres étapes dans le protocole, comme l'anticorps secondaire ou le PFA?
2.) Est-ce que vous n'avez pas besoin de perméabilisation pour obtenir ce marquage?
3.) Est-ce que vous avez fait une gamme de dilution avec cet anticorps?

Je me réjouis de votre réponse.

Read More

Question

bad signal with new batch

Read More
Answer

Je suis désolée d'apprendre que un nouveau lot d'ab76081 ne vous donne pas de bons résultats.
Veuillez trouver ci-joint le questionnaire discuté au téléphone qui nous permet de regrouper le plus d'informations le protocole que vous avez utilisé avec cet anticorps. Celui-ci nous permettra de vous fournir la meilleure assistance technique si nécessaire et aussi d'avoir les informations pour initier des investigations internes.
Si nous concluons que notre produit est fautif et ne fonctionne pas comme explicité par notre fiche technique, nous serons ravis de mettre en place l'envoi d'un tube de remplacement gratuit ou d'un remboursement (sous réserve d'une commande ultérieure de 180 jours).
Je vous remercie par avance de nous envoyer votre protocole.

Read More

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