Aquaporin-2 Antibody

Western Blot: Aquaporin-2 Antibody [NB110-74682] - Detection of Aquaporin 2 in rat kidney extracts Key: CX=Cortex, OM=Outer Medulla, IMB =Inner Medulla Base, IMP = Inner Medulla Papilla

Immunocytochemistry/Immunofluorescence: Aquaporin-2 Antibody [NB110-74682] - Staining of mpkCCD cell line

Immunohistochemistry-Paraffin: Aquaporin-2 Antibody [NB110-74682] - IHC analysis of a formalin fixed paraffin-embedded (FFPE) human kidney using 1:100 conc. of Aquaporin-2 antibody on a Bond Rx autostainer (Leica Biosystems). The assay involved 20 minutes of heat induced antigen retrieval (HIER) using 10mM sodium citrate buffer (pH 6.0) and endogenous peroxidase quenching with peroxide block. The sections were incubated with primary antibody for 30 minutes and Bond Polymer Refine Detection (Leica Biosystems) with DAB was used for signal development followed by counterstaining with hematoxylin. Whole slide scanning and capturing of representative images was performed using Aperio AT2 (Leica Biosystems). Lumenal staining of Aquaporin-2 in scattered tubules was observed. Staining was performed by Histowiz.

• Reactivity: Hu, Mu, Rt, Po, Bv, Sh
• Applications: WB, ICC/IF, IHC, IHC-Fr, IHC-P
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Concentration: 0.16 mg/ml

Aquaporin-2 Antibody Summary

• Immunogen: Synthetic peptide made to a C-terminus portion of rat Aquaporin-2 (within residues 200-300). [Swiss-Prot# P34080]
• Localization: Cell membrane
• Predicted Species: Porcine (100%), Bovine (100%), Sheep (100%). Backed by our 100% Guarantee.
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: AQP2
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • Western Blot 1:1000-1:5000
  • Immunocytochemistry/Immunofluorescence 1:100-1:200
  • Immunohistochemistry 1:100-1:200
  • Immunohistochemistry-Frozen
  • Immunohistochemistry-Paraffin 1:100-1:200
• Application Notes: This Aquaporin-2 antibody is useful for Western blot, where bands can be seen ~28 kDa for the non-glycosylated form and ~37 kDa for the glycosylated form. It is also useful for Immunocytochemistry and Immunohistochemistry on paraffin-embedded sections. Immunohistochemistry-Frozen was reported in scientific literature.

Reactivity Notes

Human, rat and mouse. 100% sequence identity with sheep, porcine, and bovine proteins.

Packaging, Storage & Formulations

• Storage: Store at 4C. Do not freeze.
• Buffer: PBS and 50% Glycerol
• Preservative: 0.05% Sodium Azide
• Concentration: 0.16 mg/ml
• Purity: Immunogen affinity purified

Alternate Names for Aquaporin-2 Antibody

• ADH water channel
• AQP-2
• AQP-CD
• aquaporin 2 (collecting duct)
• aquaporin-2
• aquaporin-CD
• Collecting duct water channel protein
• MGC34501
• Water channel protein for renal collecting duct
• water-channel aquaporin 2
• WCH-CD

Background

Aquaporin 2 (AQP2) is a membrane protein involved in plasma membrane water transport in the renal collecting duct. This antibody is targeted to a peptide sequence upstream from the phosphorylated region that, upon phosphorylation, can diminish the antibody signal in immunoblots and immunocytochemistry.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Aquaporin-2 Antibody (NB110-74682)(16)

Publications using NB110-74682

• Tanaka A, Nakamura T, Sato E et al. Different Effects of Tolvaptan in Patients with Idiopathic Membranous Nephropathy with Nephrotic Syndrome Intern. Med Jan 16 2017 [PMID: 28090051] (ICC/IF, Human)
• Martins JR, Penton D, Peyronnet R et al. Piezo1-dependent regulation of urinary osmolarity. Pflugers Arch. 2016 Mar 29 [PMID: 27023350] (WB, ICC/IF, Mouse)
• Yang CR, Raghuram V, Emamian M et al. Deep Proteomic Profiling of Vasopressin-Sensitive Collecting Duct Cells. II. Bioinformatic Analysis of Vasopressin Signaling. Am. J. Physiol., Cell Physiol. 2015 Aug 26 [PMID: 26310817] (WB, Mouse)
  
  Details:
  Aquaporin-2 antibody was used for WB analysis of lysates from experiments involving mpkCCD cells and the antibody detected ~29 kDa unglycosylated as well as ~37 kDa glycosylated forms of AQP2 protein.
• Gilbert ML, Yang L, Su T, McKnight GS. Expression of a dominant negative Protein Kinase A (PKA) mutation in kidney elicits a diabetes insipidus phenotype Am. J. Physiol. Renal Physiol. 2015 Jan 13 [PMID: 25587115] (WB, IHC, Mouse)
• Sato E, Nakamura T, Amaha M et al. Effect of Tolvaptan in Patients With Chronic Kidney Disease due to Diabetic nephropathy With Heart Failure. Int Heart J. 2014 Oct 16 [PMID: 25318553] (IHC-P, Human)
• Gao Y, Romero-Aleshire MJ, Cai Q et al. Rapamycin inhibition of mTORC1 reverses lithium-induced proliferation of renal collecting duct cells. Am J Physiol Renal Physiol. 2013 Oct [PMID: 23884148] (WB, Mouse)
• Takumida M, Takumida H, Kakigi A et al. Localization of aquaporins in the mouse vestibular end organs. Acta Otolaryngol 2013 Apr 30 [PMID: 23628076] (IHC-Fr, ICC/IF, Mouse)
• Toka HR, Al-Romaih K, Koshy JM et al. Deficiency of the Calcium-Sensing Receptor in the Kidney Causes Parathyroid Hormone-Independent Hypocalciuria J Am Soc Nephrol 2012 Sep 20 [PMID: 22997254]
• Takumida M, Kakigi A, Egami N et al. Localization of aquaporins 1, 2, and 3 and vasopressin type 2 receptor in the mouse inner ear Acta Otolaryngol 2012 Jul 6 [PMID: 22768909] (IHC, ICC/IF, Mouse)
• Yu MJ, Miller RL, Uawithya P et al. Systems-level analysis of cell-specific AQP2 gene expression in renal collecting duct. Proc Natl Acad Sci U S A;106(7):2441-6. 2009 Feb 17. [PMID: 19190182] (WB, ICC/IF, Mouse)
• Moeller HB, Knepper MA, Fenton RA. Serine 269 phosphorylated aquaporin-2 is targeted to the apical membrane of collecting duct principal cells. Kidney Int;75(3):295-303. 2009 Feb. [PMID: 18843259] (WB, IHC, ICC/IF, Mouse, Rat)
• Hoffert JD, Fenton RA, Moeller HB et al. Vasopressin-stimulated increase in phosphorylation at Ser269 potentiates plasma membrane retention of aquaporin-2. J Biol Chem;283(36):24617-27. 2008 Sep 5. [PMID: 18606813] (WB, IHC, Mouse)
• Sato T, Courbebaisse M, Ide N et al. Parathyroid hormone controls paracellular Ca(2+) transport in the thick ascending limb by regulating the tight-junction protein Claudin14. Proc. Natl. Acad. Sci. U.S.A. Apr 18 2017 [PMID: 28373577] (ICC/IF, Mouse)
• Sanz E, Evanoff R, Quintana A et al. RiboTag Analysis of Actively Translated mRNAs in Sertoli and Leydig Cells In Vivo. PLoS One 2013 Jun 11 [PMID: 23776628] (WB, IHC-Fr, Mouse)
• Stegbauer J, Gurley SB, Sparks MA et al. AT1 receptors in the collecting duct directly modulate the concentration of urine. J Am Soc Nephrol 2011 Dec [PMID: 22052052] (Mouse)

FAQs for Aquaporin-2 Antibody (NB110-74682). (Showing 1 - 2 of 2 FAQs).

1. We have a customer who is interested in catalog # NB110-74682, but he needs more information regarding the sequence if possible. The range of 200-300 covers both a predicted helical and cytoplasmic domain. Would it be possible to know if the exact sequence is contained entirely within either of these domains? He is hoping for the helical as he needs to detect on the cell surface.
   • Unfortunately for your customer, NB110-74682 recognizes the cytoplasmic domain.
2. I have a customer who is interested in antibody NB100-74682 anti-Aquaporin. They are concerned about the fact that there are only a few extracellular amino acids in the region 200-300 of AQP2. Please see below. The immunogen for anti-AQP2 is listed as aa 200 - 300 of aquaporin 2. This product is listed as having application in ICC, which requires an extracellular epitope. From looking at the structure of Aquaporin 2, it appears that only about 4 residues of aa200 - 300 are extracellular. Can you explain this? Do you know if the cells were permeabilized for the ICC image shown on your datasheet or was the antibody able to cross the membrane efficiently enough to stain without permeabilizing the cells?
   • Our immunogen for NB110-74682 is to the cytoplasmic domain of AQP2. Furthermore, your customer is incorrect that ICC requires and extracellular domain, as many antibodies that recognize an intracellular target work quite well for ICC when the cells are permeabilized. We permeabilize for all of our ICC testing and this antibody certainly requires it.

Bioinformatics

• Gene Symbol: AQP2
• Entrez:
  • 359(Human)
  • 11827(Mouse)
  • 25386(Rat)
• Uniprot:
  • P34080(Rat)