Aquaporin-2 Antibody 0.05 ml

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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
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Ordering Information: NB110-74682

  • Catalog Number
    NB110-74682
  • Displayed Format
    Unconjugated
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Aquaporin-2 Antibody Summary

Species Human, Mouse, Rat
Predicted Species
Porcine (100%), Bovine (100%), Sheep (100%). Backed by our 100% Guarantee
Tested Applications WB, ICC/IF, IHC, IHC-Fr, IHC-P
Clonality
Polyclonal
Host
Rabbit
Gene
AQP2
Purity
Immunogen affinity purified
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Aquaporin-2 Antibody Details

Immunogen
Synthetic peptide made to a C-terminus portion of rat Aquaporin-2 (within residues 200-300). [Swiss-Prot# P34080]
Localization
Cell membrane

Species Reactivity

Human, rat and mouse. 100% sequence identity with sheep, porcine, and bovine proteins.
Publications Read Publications using NB110-74682 in the following species:


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.
Positive Controls
Other Available
Formats
DyLight 405 Labeled NB110-74682V
FITC Labeled NB110-74682F
Publications
Read Publications using
NB110-74682 in the following applications:

Contact Information

Product PDFs

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Reviews

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Publications

Aquaporin-2 Antibody (NB110-74682) has been mentioned in at least 11 publications.
Earn rewards if you have published using Aquaporin-2 Antibody (NB110-74682).

Packaging, Storage & Formulations

Storage
Store at 4C. Do not freeze.
Buffer
PBS
Unit Size
0.05 ml
Concentration
0.9 mg/ml
Preservative
0.05% Sodium Azide
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.

Bioinformatics

Gene Symbol AQP2
Entrez
Uniprot

Alternate Names for Aquaporin-2 Antibody

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

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.

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Publications for Aquaporin-2 Antibody (NB110-74682) (11)

Read what people are writing who have used Aquaporin-2 Antibody (NB110-74682).

We have publications tested in 3 confirmed species: Human, Mouse, Rat.

We have publications tested in 5 applications: WB, ICC/IF, IHC, IHC-Fr, IHC-P.

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Filter By Application
WB
(6)
ICC/IF
(12)
IHC
(4)
IHC-Fr
(2)
IHC-P
(1)
All Applications
Filter By Species
Human
(1)
Mouse
(8)
Rat
(1)
All Species
Publication Applications Species
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) ICC/IF, WB, ICC/IF, ICC/IF 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 Sep 5. [PMID: 18606813] (WB, IHC, Mouse) IHC, WB 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) IHC, ICC/IF, WB, ICC/IF, ICC/IF Mouse, Rat
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) IHC, ICC/IF, ICC/IF, 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, 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) ICC/IF, IHC-Fr, ICC/IF, 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) WB, IHC-Fr 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 Oct [PMID: 23884148] (WB, Mouse) WB 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) IHC-P Human
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) IHC, WB Mouse

Product General Protocols

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

Video Protocols

WB Video Protocol
ICC/IF Video Protocol

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FAQs for Aquaporin-2 (33)

  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.
  3. In the immunohistochemistry paraffin section protocol....the PBS buffer....should it be at certain pH?
    • For IHC application, different labs cites the pH of PBS buffer in the range of 7.1 - 7.6 but most commonly used pH is pH 7.4 (this is what we use in our lab). You may use this PBS (pH 7.4) for making permeablization buffer, antibody diluent buffers and for making wash buffer. For more on the protocol that we use in our lab and for IHC-P troubleshooting suggestions, you may visit: IHC-P Protocol and IHC-P Troubleshooting.
  4. I was wondering what does Format 7C mean as far as anitbodies go?
    • 7C is a fluorophore - it is useful for FLOW. The details of the fluor are (A=425, E=500)
  5. What epitope does this monoclonal antibody recognize?
    • R&D Systems does not epitope map its antibodies. The antibody is generated using the entire immunogen listed on the datasheet. If a peptide immunogen is used, this will be indicated on the datasheet.
  6. What epitope does this polyclonal antibody recognize?
    • Polyclonal antibodies generally recognize multiple epitopes because they are generated using the entire immunogen stated on the product datasheet.
  7. What are R&D Systems matched antibody pairs?
    • R&D Systems matched antibody pairs assist in the development of an ELISA. R&D Systems has demonstrated that the paired antibodies can be used in a sandwich immunoassay to recognize the recombinant protein. We recommend that the investigator have expertise in immunoassay development before attempting to use these products. Each investigator must empirically determine the optimal concentrations for the capture and detection antibodies. The amount of antibodies supplied are not normalized to develop the same number of plates. R&D Systems recombinant cytokines are not calibrated by ELISA and therefore must be mass calibrated before use. The DuoSet ELISA Development Kit product line includes the antibodies, standard, enzyme, and protocol necessary for running an immunoassay, and can often be a better value than purchasing antibody pairs and protein standard.
  8. Can I use your antibodies for a matched pair ELISA with a standard from another company?
    • Yes, but there are potential issues to consider. There can be differences in immunological recognition or the binding between antibody and recombinant protein. This can happen if there is a difference in the folding or sequence of the recombinant protein used as the standard versus the recombinant protein used as the immunogen for the antibody. Protein folding or sequence differences in the antibody-binding region can lead to poor (or no) recognition of the standard.
  9. Can I use your antibodies for a matched pair ELISA with a standard from another company?
    • Yes, but there are potential issues to consider. There can be differences in immunological recognition or the binding between antibody and recombinant protein. This can happen if there is a difference in the folding or sequence of the recombinant protein used as the standard versus the recombinant protein used as the immunogen for the antibody. Protein folding or sequence differences in the antibody-binding region can lead to poor (or no) recognition of the standard.
  10. What grade BSA is recommend for use in ELISA and ELISpot Development Systems?
    • The grade of the BSA used has been found to be a critical component for running a successful ELISA. BSA with a minimum purity of 98% is recommended. R&D Systems recommends using the same BSAs used in development of the DuoSet; Catalog # DY995 Reagent Diluent Concentrate 2, Catalog # DY997 Reagent Diluent Concentrate 1. Millipore Probumin® Diagnostic Grade BSA (Millipore, Catalog # 82-045) is another example.
  11. What grade BSA is recommend for use in ELISA and ELISpot Development Systems?
    • The grade of the BSA used has been found to be a critical component for running a successful ELISA. BSA with a minimum purity of 98% is recommended. R&D Systems recommends using the same BSAs used in development of the DuoSet; Catalog # DY995 Reagent Diluent Concentrate 2, Catalog # DY997 Reagent Diluent Concentrate 1. Millipore Probumin® Diagnostic Grade BSA (Millipore, Catalog # 82-045) is another example.
  12. What is a direct ELISA?
    • In a direct ELISA, a plate is coated with the analyte of interest and a labeled detection antibody is used to verify the presence of the analyte.The direct ELISA may use a colorimetric, chemiluminescent, or fluorescent reporter.
  13. What is a direct ELISA?
    • In a direct ELISA, a plate is coated with the analyte of interest and a labeled detection antibody is used to verify the presence of the analyte.The direct ELISA may use a colorimetric, chemiluminescent, or fluorescent reporter.
  14. What is a sandwich ELISA?
    • A sandwich ELISA uses an immobilized capture antibody specific for the analyte of interest in a sample. After the analyte is bound to the immobilized antibody, a labeled secondary antibody specific for the analyte is used for detection. The analyte is "sandwiched" between the two antibodies. The sandwich ELISA is extremely sensitive, and the values obtained are quantitative when compared with a standard curve.
  15. What is a sandwich ELISA?
    • A sandwich ELISA uses an immobilized capture antibody specific for the analyte of interest in a sample. After the analyte is bound to the immobilized antibody, a labeled secondary antibody specific for the analyte is used for detection. The analyte is "sandwiched" between the two antibodies. The sandwich ELISA is extremely sensitive, and the values obtained are quantitative when compared with a standard curve.
  16. What is the difference between AB###, AF###, BAF###, MAB###, and BAM### antibody catalog number designations?
    • AB### designated antibodies are polyclonal antibodies that are purified using Protein A, Protein G or Ion Exchange chromatography. Consequently, the resulting purified antibody is a total IgG fraction and may contain IgG not specific for the analyte of interest. AF### designated antibodies, are antigen affinity-purified and then further purified by ion exchange or size exclusion. Therefore, AF### antibodies are IgG specific only to the antigen. Antibodies that have the designation MAB### are monoclonal antibodies. BAF### and BAM### designated antibodies are the biotinylated versions of the AF### and MAB### designated antibodies, respectively.
  17. What is the difference between AB###, AF###, BAF###, MAB###, and BAM### antibody catalog number designations?
    • AB### designated antibodies are polyclonal antibodies that are purified using Protein A, Protein G or Ion Exchange chromatography. Consequently, the resulting purified antibody is a total IgG fraction and may contain IgG not specific for the analyte of interest. AF### designated antibodies, are antigen affinity-purified and then further purified by ion exchange or size exclusion. Therefore, AF### antibodies are IgG specific only to the antigen. Antibodies that have the designation MAB### are monoclonal antibodies. BAF### and BAM### designated antibodies are the biotinylated versions of the AF### and MAB### designated antibodies, respectively.
  18. What is the rationale for using trehalose to stabilize proteins?
    • Trehalose is an effective sugar for stabilizing proteins against damage caused by freezing. It can also make the protein more resistant to moisture gain when lyophilized, resulting in a product that is less likely to precipitate when reconstituted. In addition, it has been used in approved parenteral therapeutics
  19. What is the rationale for using trehalose to stabilize proteins?
    • Trehalose is an effective sugar for stabilizing proteins against damage caused by freezing. It can also make the protein more resistant to moisture gain when lyophilized, resulting in a product that is less likely to precipitate when reconstituted. In addition, it has been used in approved parenteral therapeutics
  20. Will trehalose affect my conjugation reaction?
    • It is possible that the presence of trehalose will interfere in the successful conjugation of a protein. This will depend on the method used, and the customer should investigate this potential prior to purchasing the product.
  21. Will trehalose affect my conjugation reaction?
    • It is possible that the presence of trehalose will interfere in the successful conjugation of a protein. This will depend on the method used, and the customer should investigate this potential prior to purchasing the product.
  22. Will trehalose affect the performance of the protein or antibody in my specific application?
    • We have seen no adverse effect in our bioassays or other approved applications. However, customers are advised to run a control in their assay to determine if the concentration of trelahose in the protein or antibody formulation has any adverse effects.
  23. Will trehalose affect the performance of the protein or antibody in my specific application?
    • We have seen no adverse effect in our bioassays or other approved applications. However, customers are advised to run a control in their assay to determine if the concentration of trelahose in the protein or antibody formulation has any adverse effects.
  24. Will trehalose included in the formulation affect the animal if it is injected?
    • Trehalose is unlikely to have an effect in vivo. It has been approved as an excipient for use in human injectable drugs.The trehalose used by R&D Systems is derived from Saccharomyces cerevisiae and is determined to be at minimum 98.5% pure by HPAE.
  25. Will trehalose included in the formulation affect the animal if it is injected?
    • Trehalose is unlikely to have an effect in vivo. It has been approved as an excipient for use in human injectable drugs.The trehalose used by R&D Systems is derived from Saccharomyces cerevisiae and is determined to be at minimum 98.5% pure by HPAE.
  26. What are R&D Systems matched antibody pairs?
    • R&D Systems matched antibody pairs assist in the development of an ELISA. R&D Systems has demonstrated that the paired antibodies can be used in a sandwich immunoassay to recognize the recombinant protein. We recommend that the investigator have expertise in immunoassay development before attempting to use these products. Each investigator must empirically determine the optimal concentrations for the capture and detection antibodies. The amount of antibodies supplied are not normalized to develop the same number of plates. R&D Systems recombinant cytokines are not calibrated by ELISA and therefore must be mass calibrated before use. The DuoSet ELISA Development Kit product line includes the antibodies, standard, enzyme, and protocol necessary for running an immunoassay, and can often be a better value than purchasing antibody pairs and protein standard.
  27. What is the recommended protocol for reconstituting and aliquotting lyophilized proteins and antibodies?<br>
    • Following these guidelines for reconstituting lyophilized material will help ensure complete recovery of the protein or antibody.

      1) Tap or briefly centrifuge the vial before opening to dislodge any lyophilized material that may be dispersed on the wall or cap of the vial.

      2) Use the buffer and stock concentration recommended in the product datasheet. If you want a stock concentration that is higher than the one recommended, contact Technical Service for specific recommendations.

      3) For optimal recovery, reconstitute using room temperature buffer.

      4) After adding the buffer, re-cap the vial and invert gently by hand or place on a slow rocking platform. This will allow the reconstitution buffer to coat all the surfaces inside the vial. Do not mix by vortexing or by pipetting the material up and down.

      5) Allow the vial to sit at room temperature with gentle agitation for at least 15 minutes before aliquotting or using.

      6) Store the reconstituted protein in polypropylene or siliconized tubes. If aliquotting, it is recommended that aliquots be no smaller than 10 μL. In addition, avoid repeated freeze/thaw cycles.

  28. What is the recommended protocol for reconstituting and aliquotting lyophilized proteins and antibodies?<br>
    • Following these guidelines for reconstituting lyophilized material will help ensure complete recovery of the protein or antibody.

      1) Tap or briefly centrifuge the vial before opening to dislodge any lyophilized material that may be dispersed on the wall or cap of the vial.

      2) Use the buffer and stock concentration recommended in the product datasheet. If you want a stock concentration that is higher than the one recommended, contact Technical Service for specific recommendations.

      3) For optimal recovery, reconstitute using room temperature buffer.

      4) After adding the buffer, re-cap the vial and invert gently by hand or place on a slow rocking platform. This will allow the reconstitution buffer to coat all the surfaces inside the vial. Do not mix by vortexing or by pipetting the material up and down.

      5) Allow the vial to sit at room temperature with gentle agitation for at least 15 minutes before aliquotting or using.

      6) Store the reconstituted protein in polypropylene or siliconized tubes. If aliquotting, it is recommended that aliquots be no smaller than 10 μL. In addition, avoid repeated freeze/thaw cycles.

  29. Can this antibody be used on frozen or paraffin-embedded&nbsp; sections?
    • R&D Systems will provide support for an antibody that is validated for the IHC/ICC application in both frozen and paraffin-embedded sections unless otherwise specified on the datasheet. Should the product fail to work after contacting us for assistance, R&D Systems will offer a product credit toward a future purchase.

      If the datasheet does not have an IHC/ICC claim, we do not have any in-house or collaborative data available to support this application and we cannot guarantee results. It is up to the user to decide if they wish to use the antibody in their application.

      Customers may use the Citations tab on the product-specific web page or contact Technical Service to check for references using a product in an application or with a cell type R&D Systems has not tested in-house.
  30. Can this antibody be used on frozen or paraffin-embedded&nbsp; sections?
    • R&D Systems will provide support for an antibody that is validated for the IHC/ICC application in both frozen and paraffin-embedded sections unless otherwise specified on the datasheet. Should the product fail to work after contacting us for assistance, R&D Systems will offer a product credit toward a future purchase.

      If the datasheet does not have an IHC/ICC claim, we do not have any in-house or collaborative data available to support this application and we cannot guarantee results. It is up to the user to decide if they wish to use the antibody in their application.

      Customers may use the Citations tab on the product-specific web page or contact Technical Service to check for references using a product in an application or with a cell type R&D Systems has not tested in-house.
  31. Can this antibody be used on frozen or paraffin-embedded&nbsp; sections?
    • R&D Systems will provide support for an antibody that is validated for the IHC/ICC application in both frozen and paraffin-embedded sections unless otherwise specified on the datasheet. Should the product fail to work after contacting us for assistance, R&D Systems will offer a product credit toward a future purchase.

      If the datasheet does not have an IHC/ICC claim, we do not have any in-house or collaborative data available to support this application and we cannot guarantee results. It is up to the user to decide if they wish to use the antibody in their application.

      Customers may use the Citations tab on the product-specific web page or contact Technical Service to check for references using a product in an application or with a cell type R&D Systems has not tested in-house.
  32. Has a product ever been used for an application that is not listed on the datasheet?
    • If a specific application is not listed on the datasheet, it may mean that this product has not been tested in this application, or it may mean that in-house testing in this application did not meet R&D Systems’specifications. Please check our Citations tab to see if other researchers have published using your application, sample type and/or species. If you would like more information on whether or not an application has been tested, please contact Technical Service at (800) 343-7475.

  33. Has a product ever been used for an application that is not listed on the datasheet?
    • If a specific application is not listed on the datasheet, it may mean that this product has not been tested in this application, or it may mean that in-house testing in this application did not meet R&D Systems’specifications. Please check our Citations tab to see if other researchers have published using your application, sample type and/or species. If you would like more information on whether or not an application has been tested, please contact Technical Service at (800) 343-7475.