WNT2 antibody (AA 221-320)

Catalog No. ABIN762896

Product Details anti-WNT2 Antibody

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

Binding Specificity: AA 221-320

Reactivity: Human, Mouse

Host: Rabbit

Clonality: Polyclonal

Conjugate: This WNT2 antibody is un-conjugated

Application: Western Blotting (WB), ELISA

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 (WNT2 Products)

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