DVL1 antibody (AA 21-100)

Catalog No. ABIN670671

Product Details anti-DVL1 Antibody

Target: DVL1 (Dishevelled Segment Polarity Protein 1 (DVL1))

Binding Specificity: AA 21-100

Reactivity: Human, Mouse, Rat

Host: Rabbit

Clonality: Polyclonal

Conjugate: This DVL1 antibody is un-conjugated

Application: Western Blotting (WB), ELISA, Flow Cytometry (FACS), Immunofluorescence (Cultured Cells) (IF (cc)), Immunofluorescence (Paraffin-embedded Sections) (IF (p)), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Immunohistochemistry (Frozen Sections) (IHC (fro))

Cross-Reactivity: Human, Mouse, Rat

Purification: Purified by Protein A.

Immunogen: KLH conjugated synthetic peptide derived from human DVL1

Isotype: IgG

Application Details

Application Notes: WB 1:300-5000
ELISA 1:500-1000
FCM 1:20-100
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 #029657 (Immunohistochemistry)

by: Immunohistochemistry Core, NYU Langone

No.: #029657

Date: 04/03/2014

Lot Number: 140106

Method validated: Immunohistochemistry

Positive Control: Human kidney tubules

Negative Control: Human colon stromal tissue

Notes: While faint, signal is detectable in positive control tissue and not in negative control tissue. Note that staining is expected in colon epithelial cells, and is not expected in kidney glomeruli.

Validation Images

Figure 1: Human kidney tissue stained with anti-DVL1 (brown) and counterstained with hematoxylin.

Figure 2: Human colon tissue stained with anti-DVL1 (brown) and counterstained with hematoxylin.

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

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

Full Methods

Primary Antibody:

• Antigen: Dishevelled, Dsh Homolog 1 (Drosophila) (DVL1)
• Catalog number: ABIN670671
• Lot number: 140106

Secondary Antibody:

• Antibody: Biotinylated goat anti-rabbit/anti-mouse (Kit)
• Lot number: D07640BA

Full Protocol:

• Immunohistochemistry was performed on a Ventana NEXes automated platform; instrument manufacturer specific reagents are italicized.
• 1. Slides were preheated in convection oven at 60°C for 30 min
• 2. Deparaffinization procedure: - 3 changes of Xylene, 5 min each - 3 changes of 100% Ethanol, 3 min each - 3 changes of 95% Ethanol, 3 min each - Rinsed in distilled water, 3 changes
• 3. Heat retrieval procedure - Slides retrieved in 10.0 mM Citrate, pH6.0 in a 1000W microwave oven (~100°C) for 15 min. - Slides were allowed to cool (in citrate) for 30 min. - Slides were washed x 3 in Distilled water
• 4. NEXes instrument procedure, iView DAB paraffin protocol (*abridged*): - Slide chamber warmed to 37°C
• 5. Slides rinsed with *reaction buffer* x3
• 6. *iView Inhibitor (H2O2)* applied and incubated for 4 min
• 7. Slides rinsed with *reaction buffer*
• 8. Antibody Application - Primary antibody diluted 1:250 in PBS (100 microliter applied/slide) - Ventana Isotype control applied neat - Slides Incubated overnight at room temperature (~12 hours ~25°C)
• 9. Slides rinsed with *reaction buffer* x3
• 10. *iView Biotinylated IgG* applied and incubated for 8 min
• 11. Slides rinsed with *reaction buffer*
• 14. *iView Streptavidin-Horseradish Peroxidase* applied and incubated for 8 min
• 15. Slides rinsed with *reaction buffer*
• 16. *iView DAB/H2O2* applied and incubated for 8 min
• 17. Slides rinsed with *reaction buffer*
• 18. *iView Copper* applied and incubated for 4 min
• 19. Slides rinsed with *reaction buffer*
• 20. Slides washed in Dawn Detergent/tap water
• 21. Counterstain Procedure - Hematoxylin (Leica 560 MX) 30 sec - Slides washed in tap water, 1 min - Decolorized (10% Acetic Acid in 70% ethanol), 1 min - Slides washed in tap water, 1 min - Bluing (Austin Clear Ammonia), 1 min - Slides washed in tap water, 1 min
• 22. Dehydration/coverslipping procedure: - 3 changes of 95% Ethanol, 3 min each - 3 changes of 100% Ethanol, 3 min each - 3 changes of Xylene, 5 min each - Mounted with Permount
• 23. Imaging: Leica SCN 400F Whole Slide Scanner with Digital Image Hub and Leica Slidepath software

Experimental Notes:

• Step 1: Heated tissue 60°C for 30 minutes; manufacturer heats for 45 minutes.
• Step 2: No ethanol wash was performed during deparaffinization; manufacturer includes 1 wash of 80% ethanol for 3 minutes.
• Step 3.1: Slides were heated for 15 minutes; manufacturer provides a range of 15-20 minutes.
• Step 3.2: Slides were cooled for 30 minutes; manufacturer cools for 20 minutes.
• Step 4: Italicized reagents and incubation time are fixed instrument parameters.
• Step 5: Secondary species-specific serum block not used; manufacturer blocks with 5% normal goat serum for 2 hours.
• Step 8.1: Antibody diluted in PBS at 1:250; manufacture did not recommend diluent or dilution.
• Step 8.2.1: Primary antibody incubated at room temperature overnight; manufacturer incubates overnight 4°C with agitation.

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-DVL1 antibody (ABIN670671)

Hua, Xu, He, Jiang, Ye, Pan: "Wnt4/?-catenin signaling pathway modulates balloon-injured carotid artery restenosis via disheveled-1." in: International journal of clinical and experimental pathology, Vol. 7, Issue 12, pp. 8421-31, (2015) (PubMed).

Target Details for DVL1

Target: DVL1 (Dishevelled Segment Polarity Protein 1 (DVL1))

Alternative Name: DVL1 (DVL1 Products)

Synonyms: DVL antibody, DVL1L1 antibody, DVL1P1 antibody, Dvl antibody, mKIAA4029 antibody, dvl-1 antibody, DSH antibody, DVL-1 antibody, dvl1 antibody, dvl2l antibody, Xdsh antibody, dsh1 antibody, dishevelled segment polarity protein 1 antibody, dishevelled, dsh homolog 1 (Drosophila) antibody, microRNA 6808 antibody, dishevelled segment polarity protein 1b antibody, dishevelled segment polarity protein 1 L homeolog antibody, DVL1 antibody, Dvl1 antibody, MIR6808 antibody, dvl1b antibody, dvl1.L antibody

Background:

Synonyms: DVL, DVL1L1, DVL1P1, Segment polarity protein dishevelled homolog DVL-1, Dishevelled-1, DSH homolog 1, DVL1

Background: Participates in Wnt signaling by binding to the cytoplasmic C-terminus of frizzled family members and transducing the Wnt signal to down-stream effectors. Plays a role both in canonical and non-canonical Wnt signaling. Plays a role in the signal transduction pathways mediated by multiple Wnt genes. Required for LEF1 activation upon WNT1 and WNT3A signaling. DVL1 and PAK1 form a ternary complex with MUSK which is important for MUSK-dependent regulation of AChR clustering during the formation of the neuromuscular junction (NMJ).

Gene ID: 1855

UniProt: O14640

Pathways: WNT Signaling, Synaptic Membrane, Skeletal Muscle Fiber Development