Anti-Aqp2 Antibody Picoband™

Product Name

Anti-Aqp2 Antibody Picoband™

View all Aquaporin-2 Antibodies

SKU/Catalog Number

A00935

Size

100 µg/vial

Form

Lyophilized

Description

Boster Bio Anti-Aqp2 Antibody Picoband™ catalog # A00935. Tested in WB, FCM, ELISA applications. This antibody reacts with Mouse, Rat.

Storage & Handling

At -20°C for one year from date of receipt. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for six months. Avoid repeated freezing and thawing.

Cite This Product

Anti-Aqp2 Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A00935)

Host

Rabbit

Contents

Each vial contains 4 mg Trehalose, 0.9 mg NaCl, 0.2 mg Na2HPO4.

Clonality

Polyclonal

Clone Number

1B9

Isotype

IgG

Immunogen

E.coli-derived mouse Aqp2 recombinant protein (Position: A102-A271). Mouse Aqp2 shares 87.6% and 98.2% amino acid (aa) sequence identity with human and rat Aqp2, respectively.

*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.

Cross-reactivity

No cross reactivity with other proteins.

Reactive Species

A00935 is reactive to AQP2 in Mouse, Rat

Applications

A00935 is guaranteed for ELISA, Flow Cytometry, WB Boster Guarantee

Observed Molecular Weight

26 kDa

Calculated molecular weight

28.837kDa

Background of Aquaporin-2

AQP2 (Aquaporin 2), also called AQUAPORIN-CD, is found in the apical cell membranes of the kidney's collecting duct principal cells and in intracellular vesicles located throughout the cell. The AQP2 gene is mapped to chromosome 12q13, very close to the site of major intrinsic protein by situ hybridization. The investigators suggested that a defect in the AQP2 gene is the basis of the autosomal form of nephrogenic diabetes insipidus. The functional expression and the limited localization suggested that AQP2 is the vasopressin-regulated water channel. Using rat kidney slices and porcine kidney cells stably expressing rat Aqp2, AQP2 trafficking can be stimulated by cAMP-independent pathways that utilize nitric oxide (NO). The NO donors sodium nitroprusside (SNP) and NONOate and the NO synthase substrate L-arginine mimicked the effect of vasopressin (VP), stimulating relocation of Aqp2 from cytoplasmic vesicles to the apical plasma membrane. SNP increased intracellular cGMP rather than cAMP, and exogenous cGMP stimulated AQP2 membrane insertion. Atrial natriuretic factor, which signals via cGMP, also stimulated AQP2 translocation. AQP2 expression in kidney connecting tubules is sufficient for survival and that AQP2 expression in collecting ducts is required to regulate body water balance. The S256L substitution in the cytoplasmic tail of the Aqp2 protein prevented phosphorylation at S256 and the subsequent accumulation of Aqp2 on the apical membrane of the collecting duct principal cells.

Antibody Validation

Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.

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Reconsitution

Adding 0.2 ml of distilled water will yield a concentration of 500 µg/ml.

Assay Dilutions Recommendation

The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.

Western blot, 0.25-0.5 µg/ml, Mouse, Rat
Flow Cytometry (Fixed), 1-3 µg/1x10⁶ cells, Mouse
ELISA, 0.1-0.5 µg/ml, Mouse

Validation Images & Assay Conditions

Click image to see more details

Figure 1. Western blot analysis of Aqp2 using anti-Aqp2 antibody (A00935).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: rat kidney tissue lysates,
Lane 2: mouse kidney tissue lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-Aqp2 antigen affinity purified polyclonal antibody (Catalog # A00935) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for Aqp2 at approximately 26 kDa. The expected band size for Aqp2 is at 30 kDa.

Click image to see more details

Figure 2. Flow Cytometry analysis of RAW264.7 cells using anti-Aqp2 antibody (A00935).
Overlay histogram showing RAW264.7 cells stained with A00935 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-Aqp2 Antibody (A00935, 1 μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Gene/Protein Information For AQP2 (Source: Uniprot.org, NCBI)

Alternative 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 AQP2 AQP-CD, WCH-CD aquaporin 2 aquaporin-2|ADH water channel|AQP-2|aquaporin 2 (collecting duct)|aquaporin-CD|collecting duct water channel protein|water channel protein for renal collecting duct|water-channel aquaporin 2

For more info on AQP2, check out the AQP2 Infographic

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In this infographic, you will see the following information for AQP2: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].

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