Overview

  • Product name
  • Description
    Rabbit polyclonal to Axin 2
  • Host species
    Rabbit
  • Specificity
    Milk blocking not recommended
  • Tested applications
    Suitable for: IHC-P, IHC-Fr, ICC/IF, WB, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
    Predicted to work with: Chicken
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 800 to the C-terminus of Mouse Axin 2.

    (Peptide available as ab32196.)

  • Positive control
    • This antibody gave a positive signal in: SW480, HCT 116, LS147T whole cell lysates; mouse colon tissue lysates and AXIN2 partial recombinant tagged protein

Properties

Applications

Our Abpromise guarantee covers the use of ab32197 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
IHC-P Use a concentration of 0.1 - 1 µg/ml.
IHC-Fr Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.
WB Use a concentration of 1 µg/ml. Can be blocked with Mouse Axin 2 peptide (ab32196). Detects a band of approximately 94 kDa (predicted molecular weight: 94 kDa) in endogenous lysates. Ab32197 gave a positive signal against the tagged partial recombinant protein which has an expected molecular weight of 37 kDa.
Flow Cyt 1/50.

ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.

 

Target

  • Function
    Inhibitor of the Wnt signaling pathway. Down-regulates beta-catenin. Probably facilitate the phosphorylation of beta-catenin and APC by GSK3B.
  • Tissue specificity
    Expressed in brain and lymphoblast.
  • Involvement in disease
    Defects in AXIN2 are involved in colorectal cancer (CRC) [MIM:114500]. They appear to be specifically associated with defective mismatch repair.
    Defects in AXIN2 are the cause of oligodontia-colorectal cancer syndrome (ODCRCS) [MIM:608615]. Affected individuals manifest severe tooth agenesis and colorectal cancer or precancerous lesions of variable types.
  • Sequence similarities
    Contains 1 DIX domain.
    Contains 1 RGS domain.
  • Domain
    The tankyrase-binding motif (also named TBD) is required for interaction with tankyrase TNKS and TNKS2.
  • Post-translational
    modifications
    Probably phosphorylated by GSK3B and dephosphorylated by PP2A.
    ADP-ribosylated by tankyrase TNKS and TNKS2. Poly-ADP-ribosylated protein is recognized by RNF146, followed by ubiquitination and subsequent activation of the Wnt signaling pathway.
    Ubiquitinated by RNF146 when poly-ADP-ribosylated, leading to its degradation and subsequent activation of the Wnt signaling pathway. Deubiquitinated by USP34, deubiquitinated downstream of beta-catenin stabilization step: deubiquitination is important Wnt signaling to positively regulate beta-catenin (CTNBB1)-mediated transcription.
  • Cellular localization
    Cytoplasm.
  • Information by UniProt
  • Database links
  • Alternative names
    • Axil antibody
    • Axin like protein antibody
    • Axin-2 antibody
    • Axin-like protein antibody
    • Axin2 antibody
    • AXIN2_HUMAN antibody
    • Axis inhibition protein 2 antibody
    • Conductin antibody
    • DKFZp781B0869 antibody
    • MGC10366 antibody
    • MGC126582 antibody
    see all

Images

  • Anti-Axin 2 antibody (ab32197) at 1 µg/ml + AXIN2 partial recombinant tagged protein at 0.1 µg

    Secondary
    IRDye 680 Conjugated Goat Anti-Rabbit IgG at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size: 94 kDa



    ab32197 gave a positive signal against the tagged partial recombinant protein which has an expected molecular weight of 37 kDa.

  • ab32197 staining Axin2 in mouse colon sections. Secondary antibody conjugated to Alexa-fluor ® 488 (please see Abreview for further details)

    See Abreview

  • IHC image of ab32197 stained area of normal tissue in a human colorectal cancer FFPE section. Section was pre-treated using pressure cooker heat mediated antigen retrieval with sodium citrate buffer (pH6) for 30 seconds at 125°C. Section was incubated with ab32197 at a dilution of 1:500 for 1h at room temperature and detected using an HRP conjugated polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
  • All lanes : Anti-Axin 2 antibody (ab32197) at 1 µg/ml

    Lane 1 : SW480 (Human colon adenocarcinoma cell line) Whole Cell Lysate
    Lane 2 : Colon (Mouse) Tissue Lysate
    Lane 3 : HCT 116 (Human Colorectal Carcinoma) Whole Cell Lysate
    Lane 4 : LS147T (Colon cancer cells) (Human) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) (ab65484) at 1/3000 dilution

    Performed under reducing conditions.

    Predicted band size: 94 kDa
    Observed band size: 94 kDa
    Additional bands at: 32 kDa, 75 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 16 minutes
  • Flow Cytometry analysis of human permeabilised REH B cells labeling Axin 2 with ab32197 (red). Cells were fixed with paraformaldehyde and permeabilized with saponin, followed by incubation with 2ul of undiluted Anti-Axin 2 antibody (ab32197)  in 100µl of cells in PBS for 15 minutes at 4°C. An undiluted polyclonal goat anti-rabbit Alexa Fluor® 488 was used as the secondary antibody. Control - rabbit IgG (white).

    See Abreview

References

This product has been referenced in:
  • Ding Y  et al. Deubiquitinase inhibitor b-AP15 activates endoplasmic reticulum (ER) stress and inhibits Wnt/Notch1 signaling pathway leading to the reduction of cell survival in hepatocellular carcinoma cells. Eur J Pharmacol 825:10-18 (2018). Read more (PubMed: 29454609) »
  • Yuan L & Tian J LIN28B promotes the progression of colon cancer by increasing B-cell lymphoma 2 expression. Biomed Pharmacother 103:355-361 (2018). WB ; Human . Read more (PubMed: 29669301) »

See all 29 Publications for this product

Customer reviews and Q&As

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Application
Western blot
Loading amount
50 µg
Gel Running Conditions
Reduced Denaturing
Sample
Mouse Tissue lysate - whole (colon cancer colitis induced)
Specification
colon cancer colitis induced
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C
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Submitted Jul 25 2014

Application
Western blot
Loading amount
50 µg
Gel Running Conditions
Reduced Denaturing
Sample
Human Cell lysate - whole cell (HCT116 colorectal adenocarcinoma)
Specification
HCT116 colorectal adenocarcinoma
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C
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Submitted Jul 25 2014

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Colon)
Specification
Colon
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10 mM Citrate
Permeabilization
No
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
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Submitted Feb 13 2013

Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (mouse E10.5 embryo)
Specification
mouse E10.5 embryo
Fixative
Paraformaldehyde
Permeabilization
Yes - PBST (PBS/0.5%v/v TritonX100 (Sigma)
Blocking step
NOVOCASTRA protein block as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 25°C
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Submitted Jun 08 2012

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry free floating
Sample
Chicken Tissue sections (HH 24 trunk section)
Specification
HH 24 trunk section
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Submitted Nov 10 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (HeLa Cells)
Loading amount
20 µg
Specification
HeLa Cells
Gel Running Conditions
Non-reduced Denaturing (10% gel)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
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Submitted Mar 23 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (tooth bud)
Specification
tooth bud
Fixative
Paraformaldehyde
Antigen retrieval step
Other
Permeabilization
No
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10%
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Submitted Aug 13 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Flow Cytometry
Sample
Human Cell (REH B cell line)
Specification
REH B cell line
Fixation
Paraformaldehyde
Permeabilization
Yes - saponin
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Submitted May 18 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (colon section)
Specification
colon section
Fixative
Paraformaldehyde
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 2%
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Submitted May 06 2010

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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