AQP2 antibody (AA 171-271)

Catalog No. ABIN707576

The Rabbit Polyclonal anti-AQP2 antibody has been validated for WB, ELISA, IF (p), IF (cc), IHC (p), ICC, FACS and IHC (fro) and has been independently validated. It is suitable to detect AQP2 in samples from Human, Rat and Mouse. There are 2+ publications available.

Quick Overview for AQP2 antibody (AA 171-271) (ABIN707576)

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

Reactivity: Human, Rat, Mouse

Host: Rabbit

Clonality: Polyclonal

Conjugate: This AQP2 antibody is un-conjugated

Application: 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))

Product Details anti-AQP2 Antibody

Binding Specificity: AA 171-271

Cross-Reactivity: Human, Mouse, Rat

Predicted Reactivity: Dog

Purification: Purified by Protein A.

Immunogen: KLH conjugated synthetic peptide derived from human AQP2.

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
ICC 1:100-500

Restrictions: For Research Use only

Independent Validation (IHC)

Validation #029457 (Immunohistochemistry)

by: Immunohistochemistry Core, NYU Langone

No.: #029457

Date: 01/16/2014

Lot Number: 999994W

Method validated: Immunohistochemistry

Positive Control: - Human kidney tubules - Human breast ductal epithelium

Negative Control: - Human kidney glomeruli and stromal tissue - Human breast mesenchymal tissue

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

Validation Images

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).

Full Methods

Primary Antibody:

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

Secondary Antibody:

• 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

Experimental Notes:

• 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.

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-AQP2 antibody (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).

Target Details for AQP2

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

Alternative Name: AQP2

Background:

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.

Gene ID: 359

UniProt: P41181

Pathways: Response to Water Deprivation