DVL1 Antikörper (AA 21-100)

Produktnummer ABIN670671

Dieses Anti-DVL1-Antikörper ist ein Kaninchen Polyklonal-Antikörper zur Detektion von DVL1 in WB, ELISA, FACS, IF (cc), IF (p), IHC (p) und IHC (fro). Geeignet für Human, Maus und Ratte. Unabhängig validiert für die Verwendung in Immunohistochemistry. Dieses Primary Antibody wurde in 1 Publikation zitiert.

Kurzübersicht für DVL1 Antikörper (AA 21-100) (ABIN670671)

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

Reaktivität: Human, Maus, Ratte

Wirt: Kaninchen

Klonalität: Polyklonal

Konjugat: Dieser DVL1 Antikörper ist unkonjugiert

Applikation: 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))

Produktdetails anti-DVL1 Antikörper

Bindungsspezifität: AA 21-100

Kreuzreaktivität: Human, Maus, Ratte

Aufreinigung: Purified by Protein A.

Immunogen: KLH conjugated synthetic peptide derived from human DVL1

Isotyp: IgG

Anwendungsinformationen

Applikationshinweise: 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

Beschränkungen: Nur für Forschungszwecke einsetzbar

Independent Validation (IHC)

Validierung #029657 (Immunohistochemistry)

by: Immunohistochemistry Core, NYU Langone

No.: #029657

Datum: 03.04.2014

Chargennummer: 140106

Validierte Anwendung: Immunohistochemistry

Positivkontrolle: Human kidney tubules

Negativkontrolle: Human colon stromal tissue

Bewertung: 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.

Validierungsbilder

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.

Protokoll

Primärantikörper:

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

Sekundärantikörper:

• 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

Anmerkungen:

• 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.

Handhabung

Format: Liquid

Konzentration: 1 μg/μL

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

Konservierungsmittel: ProClin

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

Lagerung: 4 °C,-20 °C

Informationen zur Lagerung: Shipped at 4°C. Store at -20°C for one year. Avoid repeated freeze/thaw cycles.

Haltbarkeit: 12 months

Referenzen für anti-DVL1 Antikörper (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).

Details zu DVL1

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

Andere Bezeichnung: DVL1

Hintergrund:

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).

Gen-ID: 1855

UniProt: O14640

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