Aquaporin-2 Antibody - BSA Free

Immunocytochemistry/Immunofluorescence: Aquaporin-2 Antibody [NB110-74682] - HeLa cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.05% Triton X-100 in PBS for 5 minutes. The cells were incubated with anti-Aquaporin-2 Antibody NB110-74682 at 1 ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:1000 dilution for 60 minutes. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.

Immunocytochemistry/Immunofluorescence: Aquaporin-2 Antibody [NB110-74682] - HepG2 cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.05% Triton X-100 in PBS for 5 minutes. The cells were incubated with anti-Aquaporin-2 Antibody NB110-74682 at 1 ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:1000 dilution for 60 minutes. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

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

Immunohistochemistry-Paraffin: Aquaporin-2 Antibody [NB110-74682] - Analysis of 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.

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

• Reactivity: Hu, Mu, Rt, Po, Bv, Sh
• Applications: WB, ICC/IF, IHC
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Format: BSA Free
• Concentration: 1 mg/ml

Aquaporin-2 Antibody - BSA Free 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:
  • Immunocytochemistry/Immunofluorescence 1 ug/ml
  • Immunohistochemistry 1:100-1:200
  • Immunohistochemistry-Frozen reported in scientific literature
  • Immunohistochemistry-Paraffin 1:100-1:200
  • Western Blot 1 - 2 ug/ml
• Application Notes: Bands can be seen ~28 kDa for the non-glycosylated form and ~37 kDa for the glycosylated form in Western Blot.

Reactivity Notes

Use in Rat reported in scientific literature (PMID:33634832).

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS
• Preservative: 0.05% Sodium Azide
• Concentration: 1 mg/ml
• Purity: Immunogen affinity purified

Alternate Names for Aquaporin-2 Antibody - BSA Free

• 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)(30)

Publications using NB110-74682

• Breiderhoff T, Himmerkus N, Meoli L et al. Claudin-10a Deficiency Shifts Proximal Tubular Cl- Permeability to Cation Selectivity via Claudin-2 Redistribution Journal of the American Society of Nephrology : JASN 2022-01-14 [PMID: 35031570]
• Dixon EE, Wu H, Muto Y et al. Spatially Resolved Transcriptomic Analysis of Acute Kidney Injury in a Female Murine Model Journal of the American Society of Nephrology : JASN 2021-12-01 [PMID: 34853151] (IHC-Fr, Mouse)
• Grabowska M, Michalek K, Kedzierska-Kapuza K et al. The long-term effects of rapamycin-based immunosuppressive protocols on the expression of renal aquaporins 1, 2, 3 and 4 water channels in rats Histology and histopathology 2021-02-26 [PMID: 33634832] (IHC-P, Rat)
• Uchimura K, Wu H, Yoshimura Y, Humphreys BD Human Pluripotent Stem Cell-Derived Kidney Organoids with Improved Collecting Duct Maturation and Injury Modeling Cell reports 2020-12-15 [PMID: 33326782]
• Li B, Zhu C, Dong L et al. ADAM10 mediates ectopic proximal tubule development and renal fibrosis through Notch signalling J. Pathol. 2020-07-27 [PMID: 32715474] (IHC-P, Human)
• Bezerra Duarte D, Cavalcante Meneses G, Bandeira Lima D et al. Aquaporin-2 and NKCC2 expression pattern in patients with hepatosplenic schistosomiasis Trop. Med. Int. Health 2020-07-06 [PMID: 32632995]
• Nordstrom CK, Danckwardt-Lilliestrom N, Liu W, Rask-Andersen H Reversed polarizationof Na/K-ATPase-a sign of inverted transport in the human endolymphatic sac: a super-resolution structured illumination microscopy (SR-SIM) study Cell Tissue Res. 2019-11-12 [PMID: 31713726]
• Li ZL, Lv LL, Wang B et al. The profibrotic effects of MK-8617 on tubulointerstitial fibrosis mediated by the KLF5 regulating pathway FASEB J. 2019-08-26 [PMID: 31451021] (Mouse)
• Shi M, Luo W, Feng X et al. Urinary Angiotensinogen Predicts Renal Disease Activity in Lupus Nephritis Antioxid. Redox Signal. 2019-07-26 [PMID: 31264479]
• Reinke P and Kurtz A. Generating Multiple Kidney Progenitors and Cell Types from Human Pluripotent Stem Cells. Methods Mol Biol. 2019-01-01 [PMID: 30742266] (ICC/IF, Human)
• Liang C, Zhang P, Wu J et al. Zhen-wu-tang attenuates Adriamycin-induced nephropathy via regulating AQP2 and miR-92b. Biomedicine & Pharmacotherapy. 1905-07-11 [PMID: 30551379] (IHC-P, Rat)
• 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. 2017-04-18 [PMID: 28373577] (ICC/IF, Mouse)
• Tanaka A, Nakamura T, Sato E et al. Different Effects of Tolvaptan in Patients with Idiopathic Membranous Nephropathy with Nephrotic Syndrome Intern. Med 2017-01-16 [PMID: 28090051] (ICC/IF, Human)
• Martins JR, Penton D, Peyronnet R et al. Piezo1-dependent regulation of urinary osmolarity. Pflugers Arch. 2016-03-29 [PMID: 27023350] (WB, ICC/IF, Mouse)
• Pollow DP Jr, Romero-Aleshire MJ, Sanchez JN et al. ANG II-induced hypertension in the VCD mouse model of menopause is prevented by estrogen replacement during perimenopause Am J Physiol Regul Integr Comp Physiol. 2015-12-14 [PMID: 26491098] (WB, 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. 2015-08-26 [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. 2015-01-13 [PMID: 25587115] (WB, IF/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. 2014-10-16 [PMID: 25318553] (IHC-P, Human)
• Stegbauer J, Gurley SB, Sparks MA et al. AT1 receptors in the collecting duct directly modulate the concentration of urine. J Am Soc Nephrol 2011-12-01 [PMID: 22052052] (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. 2009-02-01 [PMID: 18843259] (WB, ICC/IF, IF/IHC, Rat, Mouse)
• 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. 2008-09-05 [PMID: 18606813] (WB, IF/IHC, Mouse)
• 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. 2013-10-01 [PMID: 23884148] (WB, Mouse)
• Sanz E, Evanoff R, Quintana A et al. RiboTag Analysis of Actively Translated mRNAs in Sertoli and Leydig Cells In Vivo. PLoS One 2013-06-11 [PMID: 23776628] (WB, IHC-Fr, Mouse)
• Takumida M, Takumida H, Kakigi A et al. Localization of aquaporins in the mouse vestibular end organs. Acta Otolaryngol 2013-04-30 [PMID: 23628076] (IHC-Fr, 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. 2009-02-17 [PMID: 19190182] (ICC/IF, WB, Mouse)
• Wypych A, Dunis?awska A, Grabowska M et al. Effects of three-month feeding high-fat diets with differentfatty acid composition on kidney histology and expressionof genes related to cellular stress and water-electrolytehomeostasis in mice Journal of Animal and Feed Sciences 2023-06-22 (IHC-P, Mouse)
• El-Hady E, Behairy A, Goda N et al. Comparative physiological, morphological, histological, and AQP2 immunohistochemical analysis of the Arabian camels (Camelus dromedarius) and oxen kidney: Effects of adaptation to arid environments Frontiers in Animal Science 2023-03-27 (IHC, Bovine, Camel)
• 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. 2012-11-01 [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 2012-07-06 [PMID: 22768909] (IF/IHC, ICC/IF, 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)