Overview

  • Product name

  • Description

    Mouse monoclonal to DKK1
  • Host species

    Mouse
  • Tested applications

    Suitable for: ICC/IF, WB, ELISAmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
    Predicted to work with: Rabbit, Horse, Cow, Chimpanzee, Rhesus monkey
  • Immunogen

    Recombinant fragment with tag: QPYPCAEDEE CGTDEYCASP TRGGDAGVQI CLACRKRRKR CMRHAMCCPG NYCKNGICVS SDQNHFRGEI EETITESFGN DHSTLDGYSR RTTLSSKMYH , corresponding to amino acids 81-181 of Human DKK1

  • Positive control

    • HeLa cell lysate.
  • General notes

    This product was changed from ascites to tissue culture supernatant on 05 Feb 2019. Please note that the dilutions may need to be adjusted accordingly. If you have any questions, please do not hesitate to contact our scientific support team.

Properties

Applications

Our Abpromise guarantee covers the use of ab61275 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
ICC/IF Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Detects a band of approximately 29 kDa (predicted molecular weight: 29 kDa).
ELISA Use at an assay dependent concentration.

Target

  • Function

    Antagonizes canonical Wnt signaling by inhibiting LRP5/6 interaction with Wnt and by forming a ternary complex with the transmembrane protein KREMEN that promotes internalization of LRP5/6. DKKs play an important role in vertebrate development, where they locally inhibit Wnt regulated processes such as antero-posterior axial patterning, limb development, somitogenesis and eye formation. In the adult, Dkks are implicated in bone formation and bone disease, cancer and Alzheimer disease.
  • Tissue specificity

    Placenta.
  • Sequence similarities

    Belongs to the dickkopf family.
  • Domain

    The C-terminal cysteine-rich domain mediates interaction with LRP5 and LRP6.
  • Cellular localization

    Secreted.
  • Information by UniProt
  • Database links

  • Alternative names

    • Dickkopf 1 antibody
    • Dickkopf 1 homolog antibody
    • Dickkopf 1 like antibody
    • Dickkopf homolog 1 antibody
    • Dickkopf like protein 1 antibody
    • Dickkopf related protein 1 antibody
    • Dickkopf WNT signaling pathway inhibitor 1 antibody
    • Dickkopf-1 antibody
    • Dickkopf-related protein 1 antibody
    • DKK 1 antibody
    • Dkk-1 antibody
    • Dkk1 antibody
    • DKK1_HUMAN antibody
    • hDkk 1 antibody
    • hDkk-1 antibody
    • SK antibody
    see all

Images

  • Anti-DKK1 antibody (ab61275) at 1 µg/ml + HeLa cell lysate at 25 µg

    Predicted band size: 29 kDa
    Observed band size: 29 kDa
    Additional bands at: 35 kDa. We are unsure as to the identity of these extra bands.



    This image was generated using a version of the antibody produced in ascites. 

  • ICC/IF image of ab61275 stained HeLa 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 (ab61275, 10µg/ml) overnight at +4°C. The secondary antibody (green) was ab96879 Dylight 488 goat anti-mouse 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.

    This image was generated using a version of the antibody produced in ascites. 

References

This product has been referenced in:

  • Yang J  et al. STAT1-induced upregulation of LINC00467 promotes the proliferation migration of lung adenocarcinoma cells by epigenetically silencing DKK1 to activate Wnt/ß-catenin signaling pathway. Biochem Biophys Res Commun N/A:N/A (2019). Read more (PubMed: 31027730) »
  • Tang X  et al. Knockdown of YAP inhibits growth in Hep-2 laryngeal cancer cells via epithelial-mesenchymal transition and the Wnt/ß-catenin pathway. BMC Cancer 19:654 (2019). Read more (PubMed: 31269911) »
See all 8 Publications for this product

Customer reviews and Q&As

1-6 of 6 Abreviews or Q&A

Answer

Thank you for contacting us.

I can confirm that the positive control is not included into an order of the DKK antibody ab61275.

However, we do offer two products of HeLa cell lysate: ab29545 (tested for WB application) and ab150035 that can be used as positive controls for WB. For further information about both products use the provided links below.

ab29545
https://www.abcam.com/index.html?datasheet=29545 (or use the following: https://www.abcam.com/index.html?datasheet=29545).


ab150035
https://www.abcam.com/index.html?datasheet=150035 (or use the following: https://www.abcam.com/index.html?datasheet=150035).


I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

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Question
Answer

Yes the replacement antibody would be free.

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Question
Answer

Thanks for your email.

You do not need to return the vial of ab61275. I have found your order on our system. I just want to confirm if you would like another vial of ab61275 or a different primary antibody.

Thank you!

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Answer

Thanks for your reply.

It does seem like there is a problem with this vial of ab61275. If you have purchased this antibody within the last 6 months or so I would be happy to send you a replacement of any other primary antibody.

In your reply, please include your Abcam order reference number or PO number so I can process your request.

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Question

Dear Abcam,
I have fill in the questions. Can we return it? Thank you very much.
1) Abcam product code ab
ab61275
2) Abcam order reference number or product batch number
GR64429
3) Description of the problem
There is no specific band
4) Sample preparation:
Split with RIPA Buffer
Type of sample (whole cell lysates, fraction, recombinant protein…):
whole cell lysates
Lysis buffer
RIPA Buffer with protease inhibitors
Protease inhibitors:
cocktail
Phosphatase inhibitors
DTT Na3NO4 NaF
Reducing agent
Boiling for ≥5 min? yes/no
Yes
Protein loaded ug/lane or cells/lane
10 μg
Positive control
DKK1-pCDNA3.1
Negative control
Cell transfected with empty vector
5) Percentage of gel
10%
Type of membrane
PVDF
Protein transfer verified
Blocking agent and concentration
5% non-fat milk
Blocking time
1 hour
Blocking temperature
RT
6) Primary antibody (If more than one was used, describe in “additional notes”) :
Anti-DKK1
Concentration or dilution
1:500 with 5% milk
Diluent buffer
TBST
Incubation time
Overnight
Incubation temperature:
4℃
7) Secondary antibody:
Anti-Mouse
Species:
Reacts against:
mouse
Concentration or dilution
1:5000
Diluent buffer
5% milk
Incubation time
2hours
Incubation temperature:
RT
Fluorochrome or enzyme conjugate:
Ecl
8) Washing after primary and secondary antibodies:
Buffer
TBST
Number of washes
3
9)Detection method
Ecl
10) How many times have you run this staining?
2
Do you obtain the same results every time?
Yes
What steps have you altered to try and optimize the use of this antibody?
The dilution of antibody from 1:100 to 1:1000
Document attachment: Attaching images of your blot is strongly recommended and can greatly speed up our investigation of your problem.

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Answer

Thanks so much for passing along the additional protocol details.

Just one more question... Have you confirmed expression of this recombinant protein by any other means, such as by western blot detection via an epitope tag or staining of the cell extracts via coomassie blue?

Thanks again for your troubleshooting efforts!

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Answer

Thank you for your enquiry. I am sorry to hear this vial of ab61275 is giving you trouble.I will be able to help you more efficiently if you don't mind providing some additional detailsregarding your protocol and sample preparation in the space below. If this is a case where the antibody is the problem I am happy to provide a replacement, credit or refund.

Thanks so much for the additional information!

=====



1) Abcam product code ab



2) Abcam order reference number or product batch number



3) Description of the problem



4) Sample preparation:

Type of sample (whole cell lysates, fraction, recombinant protein…):

Lysis buffer

Protease inhibitors:

Phosphatase inhibitors

Reducing agent

Boiling for ≥5 min? yes/no

Protein loaded ug/lane or cells/lane

Positive control

Negative control



5) Percentage of gel

Type of membrane

Protein transfer verified

Blocking agent and concentration

Blocking time

Blocking temperature



6) Primary antibody (If more than one was used, describe in “additional notes”) :

Concentration or dilution

Diluent buffer

Incubation time

Incubation temperature:



7) Secondary antibody:

Species:

Reacts against:

Concentration or dilution

Diluent buffer

Incubation time

Incubation temperature:

Fluorochrome or enzyme conjugate:



8) Washing after primary and secondary antibodies:

Buffer

Number of washes



9)Detection method

10) How many times have you run this staining?

Do you obtain the same results every time?

What steps have you altered to try and optimize the use of this antibody?



Document attachment: Attaching images of your blot is strongly recommended and can greatly speed up our investigation of your problem.





Please send this questionnaire by e-mail by cutting and pasting the text in your e-mail (no attachments other than images) to technical@abcam.com.

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