Overview

  • Product name
  • Description
    Rabbit polyclonal to Wnt1
  • Host species
    Rabbit
  • Tested applications
    Suitable for: ELISA, ICC/IF, IHC-P, IHC-Fr, WBmore details
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Rat
  • Immunogen

    Synthetic peptide within Human Wnt1 aa 50-150 (N terminal). The exact sequence is proprietary.
    Database link: P04628

  • Positive control
    • Breast carcinoma. NIH/3T3 cell lysate.

Properties

Applications

Our Abpromise guarantee covers the use of ab15251 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
ELISA Use at an assay dependent concentration.
ICC/IF Use a concentration of 1 µg/ml.
IHC-P 1/100.
IHC-Fr Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Predicted molecular weight: 41 kDa.

Target

  • Function
    Ligand for members of the frizzled family of seven transmembrane receptors. In some developmental processes, is also a ligand for the coreceptor RYK, thus triggering Wnt signaling. Probable developmental protein. May be a signaling molecule important in CNS development. Is likely to signal over only few cell diameters.
  • Sequence similarities
    Belongs to the Wnt family.
  • Cellular localization
    Secreted > extracellular space > extracellular matrix.
  • Information by UniProt
  • Database links
  • Alternative names
    • BMND16 antibody
    • INT1 antibody
    • OI15 antibody
    • oncogene Int1 antibody
    • Proto oncogene protein Wnt 1 antibody
    • Proto-oncogene Int-1 homolog antibody
    • Proto-oncogene Wnt-1 antibody
    • Wingless type MMTV integration site family member 1 antibody
    • wingless-type MMTV integration site family, member 1 (oncogene INT1) antibody
    • Wnt 1 antibody
    • wnt1 antibody
    • WNT1_HUMAN antibody
    see all

Images

  • Anti-Wnt1 antibody (ab15251) at 1/25 dilution + NIH3T3 cell lysate

    Predicted band size: 41 kDa
    Observed band size: 41 kDa

  • ab15251 staining Wnt1 in breast carcinoma by Immunohistochemistry (FFPE-sections).

  • ICC/IF image of ab15251 stained mouse 3T3 cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab15251, 1µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 WGA was used to label plasma membranes (red).

  • Breast carcinoma cells stained with ab15251 (Immunohistochemistry, paraffin sections);

References

This product has been referenced in:
  • Linde N  et al. Macrophages orchestrate breast cancer early dissemination and metastasis. Nat Commun 9:21 (2018). Read more (PubMed: 29295986) »
  • Li X  et al. Tunicamycin inhibits progression of glioma cells through downregulation of the MEG-3-regulated wnt/ß-catenin signaling pathway. Oncol Lett 15:8470-8476 (2018). Read more (PubMed: 29805584) »
See all 41 Publications for this product

Customer reviews and Q&As

1-10 of 13 Abreviews or Q&A

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
ELISA
Sample
Human Recombinant protein (CDK8/CycC)
Specification
CDK8/CycC
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 0.1% · Temperature: 22°C
Type
Direct

Abcam user community

Verified customer

Submitted Mar 14 2013

Answer

Thank you for contacting us.

It may be, as we discussed, that the PFA fixation of the the frozen tissue is the reason the immunostain is not working. I am attaching three papers that deal with antigen retrieval for IHC of frozen tissue. Two of the papers have protocols and the other is a review of antigen retrieval. Section 4.6 of the review by D'Amico is the part of the review that I think is relevant to your frozen tissue.

Since you were able to see a positive stain with the paraffin-embedded tissue without antigen retrieval, it is possible that the heat of the paraffin-embedding itself was sufficient to reverse some of the cross-linkages that were created by fixation of the tissue. That is why I am suggesting that heat-mediated retrieval of the frozen tissue sections may help develop a stain, even thought the paraffin-embedded tissue did not require a heat-mediated retrieval. I suggest trying the protocol I mentioned: Heat 10mM sodium citrate, pH 6.0, to 90C and immerse the slides in a rack in this for 30 minutes. Maintain the temperature at 90C by keeping the container with the slides in a waterbath. After 30 minutes, allow the buffer to cool for another 30 minutes, then rinse the slides in PBS, room temperature, and continue with the blocking step and the primary antibody.

You should test one slide first to see if this damages the section, before trying with many slides.

You mentioned that the tissue for the frozen samples were fixed for two hours in PFA. Was this also the length of time that the paraffin-embedded tissues were fixed, before embedding? If the tissue that was embedded in paraffin was fixed for a a longer time, then there is also the possibility that the frozen tissue samples were under-fixed.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Loading amount
35 µg
Sample
Mouse Cell lysate - whole cell (primary mouse pancreatic cancer cell lines)
Specification
primary mouse pancreatic cancer cell lines
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Dec 15 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (PanIN lesions Pancreas)
Specification
PanIN lesions Pancreas
Fixative
Acetone
Permeabilization
Yes - PBS + Triton 0.025%
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Dec 06 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (CaPan1 and Panc1)
Loading amount
35 µg
Specification
CaPan1 and Panc1
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Dec 04 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (pancreatic tumor)
Specification
pancreatic tumor
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrate
Permeabilization
Yes - PBST
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Oct 04 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Pancreatic tumor)
Specification
Pancreatic tumor
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrate
Permeabilization
Yes - PBST
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Oct 04 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Cell lysate - whole cell (Podocytes)
Loading amount
30 µg
Specification
Podocytes
Treatment
5ng/ml TGF-beta 5min to 4hr
Gel Running Conditions
Reduced Denaturing (85 gel)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Feb 24 2009

Answer

Thanks again for your email. No, the cells were not transfected with Wnt1. The lysate was used to detect the endogenous protein. If you have any additional questions, just let us know!

Read More

1-10 of 13 Abreviews or Q&A

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