AQP2 antibody (AA 171-271)

Catalog No. ABIN707576

Product Details anti-AQP2 Antibody

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

Binding Specificity: AA 171-271

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)), Immunocytochemistry (ICC), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Flow Cytometry (FACS), Immunohistochemistry (Frozen Sections) (IHC (fro))

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 (AQP2 Products)

Synonyms: AQP-CD antibody, WCH-CD antibody, cph antibody, jpk antibody, AQP-2 antibody, aquaporin-2 antibody, aquaporin 2 antibody, aquaporin 2 (collecting duct) antibody, Aqp2 antibody, AQP2 antibody

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