WNT2 antibody (AA 221-320)

Catalog No. ABIN762896

There is 1 publication for this product available. The Rabbit Polyclonal anti-WNT2 antibody is suitable to detect WNT2 in samples from Human and Mouse. It has been validated for WB and ELISA. The antibody has been independently validated.

Quick Overview for WNT2 antibody (AA 221-320) (ABIN762896)

Target: WNT2 (Wingless-Type MMTV Integration Site Family Member 2 (WNT2))

Reactivity: Human, Mouse

Host: Rabbit

Clonality: Polyclonal

Conjugate: This WNT2 antibody is un-conjugated

Application: Western Blotting (WB), ELISA

Product Details anti-WNT2 Antibody

Binding Specificity: AA 221-320

Cross-Reactivity: Human, Mouse

Purification: Purified by Protein A.

Immunogen: KLH conjugated synthetic peptide derived from human WNT2

Isotype: IgG

Application Details

Application Notes: WB 1:300-5000
ELISA 1:500-1000
IHC-P 1:200-400
IHC-F 1:100-500
IF(IHC-P) 1:50-200
IF(IHC-F) 1:50-200
IF(ICC) 1:50-200

Restrictions: For Research Use only

Independent Validation (IHC)

Validation #029629 (Immunohistochemistry)

by: Histopathology and Tissue Shared Resource, Georgetown Lombardi Comprehensive Cancer Center

No.: #029629

Date: 03/16/2014

Lot Number: 131105

Method validated: Immunohistochemistry

Positive Control: Human glioma tissue

Negative Control: Human brain

Notes: Signal was detected in positive control tissue and not in negative control tissue.

Validation Images

Figure 1: Human brain glioma tissue stained with anti-Wnt2 (brown) and counterstained with hematoxylin (blue).

Figure 2: Human normal brain tissue stained with anti-Wnt2 (brown) and counterstained with hematoxylin (blue).

Figure 3: Human brain glioma tissue stained with isotype control antibody (brown) and counterstained with hematoxylin (blue).

Figure 4: Human brain glioma tissue stained with secondary only (brown) and counterstained with hematoxylin (blue).

Figure 5: Human brain tissue with Parkinson's morphology stained with anti-Wnt2 (brown) and counterstained with hematoxylin (blue).

Full Methods

Primary Antibody:

• Antigen: Wingless-Type MMTV Integration Site Family Member 2 (WNT2) (N-Term)
• Catalog number: ABIN762896
• Lot number: 131105

Secondary Antibody:

• Antibody: Envision Plus Horse Radish Peroxidase conjugated anti-rabbit antibody
• Batch number: 10082183

Full Protocol:

• Immunohistochemistry was performed by hand.
• Sections were de-paraffinized on a Leica Autostainer in Xylenes (1X 5 min, 2X 2 min), and rehydrated through Ethanols (2X 100% for 5 min each, 95% 2X 2 min each, 80%, 70%) to running tap water. - Sections were heated to 98°C for 20 min in 10 mM Sodium Citrate buffer pH 6.0 for antigen retrieval, then moved to RT in the same buffer for 20 min.
• Sections were rinsed in de-ionized water for 5 min at RT.
• Sections were blocked in Hydrogen Peroxide (Fisher, H325-500) for 30 min at RT.
• Sections were rinsed in Tris Buffered Saline with 0.5% Tween-20 (TBST) 2 times for 5 min at RT.
• Sections were blocked in 5% Normal Goat Serum for 2 h at RT.
• Sections were incubated with primary antibody diluted 1:250 in TBST overnight at 4°C.
• Sections were rinsed in TBST for 5 min 2X at RT.
• Sections were incubated with Envision Plus anti-Rabbit-Horse Radish Peroxidase conjugated Polymer for 2 h at RT.
• Sections were rinsed in TBST for 5 min 2X at RT.
• Sections were incubated with DAB chromogenic substrate (DAKO, K348) for 10 min at RT.
• Sections were washed x 1 in Distilled Water.
• Sections were counterstained with 1:9 dilution of Harris Hematoxylin (Fisher, SH30-500D) for 2 min.
• Sections were washed x 1 in Distilled Water.
• Sections were blued in Ammonium Hydroxide for 1 min.
• Sections were washed x 1 in Distilled Water.
• Sections were dehydrated through graded alcohols, mounted in Acrymount and photographed on an Olympus DX61 microscope with DP70 camera using DP Controller and DP Manager Software.

Experimental Notes:

• The company recommends between 1:100 – 1:500 dilution factor; in our hands, 1:250 was overstained and we would recommend using a lower concentration of the antibody.
• The “normal” brain section we used proved to be from an individual who had morphological features of Parkinson’s Disease. The area shown in Figure 2 is from the morphologically normal part of the brain. The area with Parkinson’s morphology reacted strongly with the anti-wnt2 antibody (Figure 5).

Handling

Format: Liquid

Concentration: 1 μg/μL

Buffer: 0.01M TBS( pH 7.4) with 1 % BSA, 0.02 % Proclin300 and 50 % Glycerol.

Preservative: ProClin

Precaution of Use: This product contains ProClin: a POISONOUS AND HAZARDOUS SUBSTANCE, which should be handled by trained staff only.

Storage: 4 °C,-20 °C

Storage Comment: Shipped at 4°C. Store at -20°C for one year. Avoid repeated freeze/thaw cycles.

Expiry Date: 12 months

References for anti-WNT2 antibody (ABIN762896)

Król, Mucha, Majchrzak, Homa, Bulkowska, Majewska, Gajewska, Pietrzak, Perszko, Romanowska, Paw?owski, Manuali, Hellmen, Motyl: "Macrophages mediate a switch between canonical and non-canonical Wnt pathways in canine mammary tumors." in: PLoS ONE, Vol. 9, Issue 1, pp. e83995, (2014) (PubMed).

Target Details for WNT2

Target: WNT2 (Wingless-Type MMTV Integration Site Family Member 2 (WNT2))

Alternative Name: WNT2

Background:

Synonyms: IRP, INT1L1, Protein Wnt-2, Int-1-like protein 1, Int-1-related protein, WNT2

Background: Ligand for members of the frizzled family of seven transmembrane receptors. Probable developmental protein. May be a signaling molecule which affects the development of discrete regions of tissues. Is likely to signal over only few cell diameters.

Gene ID: 7472

UniProt: P09544

Pathways: WNT Signaling