Aquaporin-2 Antibody - BSA Free

Novus Biologicals | Catalog # NB110-74682

Novus Biologicals
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Key Product Details

Species Reactivity

Validated:

Human, Mouse, Rat

Cited:

Human, Mouse, Rat, Porcine, Bovine, Camel

Predicted:

Bovine (100%), Porcine (100%), Sheep (100%). Backed by our 100% Guarantee.

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Immunocytochemistry/ Immunofluorescence

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Immunocytochemistry/ Immunofluorescence, IF/IHC

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free
View all Aquaporin-2 Primary Antibodies »

Product Specifications

Immunogen

Synthetic peptide made to a C-terminus portion of rat Aquaporin-2 (within residues 200-271). [Swiss-Prot# P34080]

Reactivity Notes

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

Localization

Cell membrane

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Scientific Data Images for Aquaporin-2 Antibody - BSA Free

Immunocytochemistry/ Immunofluorescence: Aquaporin-2 Antibody - BSA Free [NB110-74682]

Immunocytochemistry/ Immunofluorescence: Aquaporin-2 Antibody - BSA Free [NB110-74682]

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 - BSA Free [NB110-74682]

Immunocytochemistry/ Immunofluorescence: Aquaporin-2 Antibody - BSA Free [NB110-74682]

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 AntibodyBSA Free [NB110-74682]

Western Blot: Aquaporin-2 AntibodyBSA Free [NB110-74682]

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 - BSA Free [NB110-74682]

Immunohistochemistry-Paraffin: Aquaporin-2 Antibody - BSA Free [NB110-74682]

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 - BSA Free [NB110-74682]

Immunocytochemistry/ Immunofluorescence: Aquaporin-2 Antibody - BSA Free [NB110-74682]

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

Applications for Aquaporin-2 Antibody - BSA Free

Application
Recommended Usage

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.

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

PBS

Format

BSA Free

Preservative

0.02% Sodium Azide

Concentration

1 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Store at -20C long term. Avoid freeze-thaw cycles.

Background: Aquaporin-2

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.

Alternate Names

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

Entrez Gene IDs

359 (Human); 11827 (Mouse); 25386 (Rat)

Gene Symbol

AQP2

UniProt

P34080 (Rat)

Additional Aquaporin-2 Products

Product Documents for Aquaporin-2 Antibody - BSA Free

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Download SDS

Product Specific Notices for Aquaporin-2 Antibody - BSA Free

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.

Citations for Aquaporin-2 Antibody - BSA Free

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Protocols

View specific protocols for Aquaporin-2 Antibody - BSA Free (NB110-74682):

Immunocytochemistry Protocol

Culture cells to appropriate density in 35 mm culture dishes or 6-well plates.

1. Remove culture medium and wash the cells briefly in PBS. Add 10% formalin to the dish and fix at room temperature for 10 minutes.
2. Remove the formalin and wash the cells in PBS.
3. Permeablize the cells with 0.1% Triton X100 or other suitable detergent for 10 min.
4. Remove the permeablization buffer and wash three times for 10 minutes each in PBS. Be sure to not let the specimen dry out.
5. To block nonspecific antibody binding, incubate in 10% normal goat serum from 1 hour to overnight at room temperature.
6. Add primary antibody at appropriate dilution and incubate overnight at 4C.
7. Remove primary antibody and replace with PBS. Wash three times for 10 minutes each.
8. Add secondary antibody at appropriate dilution. Incubate for 1 hour at room temperature.
9. Remove secondary antibody and replace with PBS. Wash three times for 10 minutes each.
10. Counter stain DNA with DAPi if required.

Immunohistochemistry-Paraffin Embedded Sections

Antigen Unmasking:
Bring slides to a boil in 10 mM sodium citrate buffer (pH 6.0) then maintain at a sub-boiling temperature for 10 minutes. Cool slides on bench-top for 30 minutes (keep slides in the sodium citrate buffer at all times).

Staining:
1. Wash sections in deionized water three times for 5 minutes each.
2. Wash sections in PBS for 5 minutes.
3. Block each section with 100-400 ul blocking solution (1% BSA in PBS) for 1 hour at room temperature.
4. Remove blocking solution and add 100-400 ul diluted primary antibody. Incubate overnight at 4 C.
5. Remove antibody solution and wash sections in wash buffer three times for 5 minutes each.
6. Add 100-400 ul HRP polymer conjugated secondary antibody. Incubate 30 minutes at room temperature.
7. Wash sections three times in wash buffer for 5 minutes each.
8. Add 100-400 ul DAB substrate to each section and monitor staining closely.
9. As soon as the sections develop, immerse slides in deionized water.
10. Counterstain sections in hematoxylin.
11. Wash sections in deionized water two times for 5 minutes each.
12. Dehydrate sections.
13. Mount coverslips.


Western Blot Protocol

1. Perform SDS-PAGE on samples to be analyzed, loading 10-25 ug of total protein per lane.
2. Transfer proteins to PVDF membrane according to the instructions provided by the manufacturer of the membrane and transfer apparatus.
3. Stain the membrane with Ponceau S (or similar product) to assess transfer success, and mark molecular weight standards where appropriate.
4. Rinse the blot TBS -0.05% Tween 20 (TBST).
5. Block the membrane in 5% Non-fat milk in TBST (blocking buffer) for at least 1 hour.
6. Wash the membrane in TBST three times for 10 minutes each.
7. Dilute primary antibody in 2% Non-fat milk in TBST and incubate overnight at 4C with gentle rocking.
8. Wash the membrane in TBST three times for 10 minutes each.
9. Incubate the membrane in diluted HRP conjugated secondary antibody in blocking buffer (as per manufacturer's instructions) for 1 hour at room temperature.
10. Wash the blot in TBST three times for 10 minutes each (this step can be repeated as required to reduce background).
11. Apply the detection reagent of choice in accordance with the manufacturer's instructions.

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

FAQs for Aquaporin-2 Antibody - BSA Free

Showing  1 - 2 of 2 FAQs Showing All

  • Q: 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?

    A: 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.

  • Q: 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.

    A: Unfortunately for your customer, NB110-74682 recognizes the cytoplasmic domain.

  • Q: 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?

    A: 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.

  • Q: 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.

    A: Unfortunately for your customer, NB110-74682 recognizes the cytoplasmic domain.

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