This antibody gave a positive signal in WB within Rat Kidney tissue lysate as well as giving a positive result in IHC in the following FFPE tissue: Rat normal kidney.
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pH: 7.40 Preservative: 0.02% Sodium azide Constituent: PBS Note: Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 1 µg/ml. Detects a band of approximately 37 kDa (predicted molecular weight: 29 kDa).
Use at an assay dependent concentration.
Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Forms a water-specific channel that provides the plasma membranes of renal collecting duct with high permeability to water, thereby permitting water to move in the direction of an osmotic gradient.
Expressed in renal collecting tubules.
Diabetes insipidus, nephrogenic, autosomal
Belongs to the MIP/aquaporin (TC 1.A.8) family.
Aquaporins contain two tandem repeats each containing three membrane-spanning domains and a pore-forming loop with the signature motif Asn-Pro-Ala (NPA).
Ser-256 phosphorylation is necessary and sufficient for expression at the apical membrane. Endocytosis is not phosphorylation-dependent.
Apical cell membrane. Basolateral cell membrane. Cytoplasmic vesicle membrane. Golgi apparatus, trans-Golgi network membrane. Shuttles from vesicles to the apical membrane. Vasopressin-regulated phosphorylation is required for translocation to the apical cell membrane. PLEKHA8/FAPP2 is required to transport AQP2 from the TGN to sites where AQP2 is phosphorylated.
IHC image of Aquaporin 2 (phospho S261) staining in Rat normal kidney formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab110418, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
Western blot - Anti-Aquaporin 2 (phospho S261) antibody (ab110418)
Anti-Aquaporin 2 (phospho S261) antibody (ab110418) at 1 µg/ml + Kidney (Rat) Tissue Lysate at 20 µg
Secondary Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution Developed using the ECL technique
Exposure time : 16 minutesAquaporin 2 contains a potential glycosylation site (SwissProt) which may explain its migration at a higher molecular weight than predicted.
The predicted molecular weight of Aquaporin 2 is 29 kDa (SwissProt), however we expect to observe a banding pattern around 37 kDa. Abcam welcomes customer feedback and would appreciate any comments regarding this product and the data presented above.
ab110418 was tested using an Indirect ELISA approach. The wells were coated with peptide (1µg/ml at 100µl/well) overnight at 4°C, followed by a 5% BSA blocking step for 1 hour at room temperature. The primary Ab was then added at a dilution range of 1- 0.00025µg/ml (100µl/well) for 1hr at room temperature. A HRP-conjugated anti-rabbit IgG (heavy and light chain) was used as a secondary antibody at 1:20,000 dilution for 1hr at room temperature.