AQP2 Antikörper (AA 171-271)

Produktnummer ABIN707576

Der Kaninchen Polyklonal Anti-AQP2-Antikörper wurde für WB, ELISA, IF (p), IF (cc), IHC (p), ICC, FACS und IHC (fro) validiert und wurde unabhängig validiert. Er ist geeignet, AQP2 in Proben von Human, Ratte und Maus zu detektieren. Es sind 2+ Publikationen verfügbar.

Kurzübersicht für AQP2 Antikörper (AA 171-271) (ABIN707576)

Target: AQP2 (Aquaporin 2 (Collecting Duct) (AQP2))

Reaktivität: Human, Ratte, Maus

Wirt: Kaninchen

Klonalität: Polyklonal

Konjugat: Dieser AQP2 Antikörper ist unkonjugiert

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

Produktdetails anti-AQP2 Antikörper

Bindungsspezifität: AA 171-271

Kreuzreaktivität: Human, Maus, Ratte

Homologie: Dog

Aufreinigung: Purified by Protein A.

Immunogen: KLH conjugated synthetic peptide derived from human AQP2.

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
ICC 1:100-500

Beschränkungen: Nur für Forschungszwecke einsetzbar

Independent Validation (IHC)

Validierung #029457 (Immunohistochemistry)

by: Immunohistochemistry Core, NYU Langone

No.: #029457

Datum: 16.01.2014

Chargennummer: 999994W

Validierte Anwendung: Immunohistochemistry

Positivkontrolle: - Human kidney tubules - Human breast ductal epithelium

Negativkontrolle: - Human kidney glomeruli and stromal tissue - Human breast mesenchymal tissue

Bewertung: Strong signal was observed in positive control tissues and not in negative control tissues.

Validierungsbilder

Figure 1: Human kidney stained with AQP2 (brown).

Figure 2. Human kidney stained with isotype control antibody (brown).

Figure 3: Human kidney stained only with secondary antibody (primary antibody omitted) (brown).

Figure 4: Human breast stained with AQP2 (brown).

Figure 5. Human breast stained with isotype control antibody (brown).

Figure 6: Human breast stained only with secondary antibody (primary antibody omitted) (brown).

Protokoll

Primärantikörper:

• Antibody: Aquaporin 2
• Catalog number: ABIN707576
• Batch number: 999994W

Sekundärantikörper:

• Antibody: Biotinylated goat anti-rabbit/anti-mouse (Kit)
• Lot number: D05923BA

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 µL 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*
• 12. *iVIEW Streptavidin-Horseradish Peroxidase* applied and incubated for 8 min
• 13. Slides rinsed with *reaction buffer*
• 14. *iVIEW DAB/H2O2* applied and incubated for 8 min
• 15. Slides rinsed with *reaction buffer*
• 16. *iVIEW Copper* applied and incubated for 4 min
• 17. Slides rinsed with *reaction buffer*
• 18. Slides washed in Dawn Detergent/tap water
• 19. Counterstain Procedure - Hematoxylin (Leica 560 MX) 30 seconds - 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
• 20. 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
• 21. Imaging: Leica SCN 400F Whole Slide Scanner with Digital Image Hub and Leica Slidepath software

Anmerkungen:

• Deviations from protocol/procedure supplied by manufacturer.
• 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 heated for 15 minutes; manufacturer provides a range of 15-20 minutes.
• Step 3.2: Slides 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.

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-AQP2 Antikörper (ABIN707576)

Lazzeri, Angelotti, Peired, Conte, Marschner, Maggi, Mazzinghi, Lombardi, Melica, Nardi, Ronconi, Sisti, Antonelli, Becherucci, De Chiara, Guevara, Burger, Schaefer, Annunziato, Anders, Lasagni et al.: "Endocycle-related tubular cell hypertrophy and progenitor proliferation recover renal function after acute kidney injury. ..." in: Nature communications, Vol. 9, Issue 1, pp. 1344, (2018) (PubMed).

Details zu AQP2

Target: AQP2 (Aquaporin 2 (Collecting Duct) (AQP2))

Andere Bezeichnung: AQP2

Hintergrund:

Synonyms: AQP-CD, WCH-CD, Aquaporin-2, AQP-2, ADH water channel, Aquaporin-CD, Collecting duct water channel protein, Water channel protein for renal collecting duct, AQP2

Background: Forms a water-specific channel that provides the plasma membranes of renal collecting duct with high permeability to water, thereby permitting water to move in the direction of an osmotic gradient.

Gen-ID: 359

UniProt: P41181

Pathways: Response to Water Deprivation