The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 1 µg/ml. Predicted molecular weight: 31 kDa.Can be blocked with Aquaporin 3 peptide (ab195690). The detection limit of ab125219 is approximately 1ng/lane under non-reducing and reducing conditions.
Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Use a concentration of 1 µg/ml.
Use a concentration of 5 µg/ml.
Use at an assay dependent concentration. ab171870-Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.
Use a concentration of 0.5 - 1 µg/ml.
FunctionWater channel required to promote glycerol permeability and water transport across cell membranes. Acts as a glycerol transporter in skin and plays an important role in regulating SC (stratum corneum) and epidermal glycerol content. Involved in skin hydration, wound healing, and tumorigenesis. Provides kidney medullary collecting duct with high permeability to water, thereby permitting water to move in the direction of an osmotic gradient. Slightly permeable to urea and may function as a water and urea exit mechanism in antidiuresis in collecting duct cells. It may play an important role in gastrointestinal tract water transport and in glycerol metabolism.
Tissue specificityWidely expressed in epithelial cells of kidney (collecting ducts) and airways, in keratinocytes, immature dendritic cells and erythrocytes. Isoform 2 is not detectable in erythrocytes at the protein level.
Sequence similaritiesBelongs to the MIP/aquaporin (TC 1.A.8) family.
DomainAquaporins contain two tandem repeats each containing three membrane-spanning domains and a pore-forming loop with the signature motif Asn-Pro-Ala (NPA).
Cellular localizationBasolateral cell membrane. In collecting ducts of kidney.
Western blot - Anti-Aquaporin 3 antibody (ab125219)
All lanes : Anti-Aquaporin 3 antibody (ab125219) at 0.5 µg/ml
Lane 1 : Rat Kidney Tissue Lysate at 50 µg Lane 2 : Rat Lung Tissue Lysate at 50 µg Lane 3 : Mouse Kidney Tissue Lysate at 50 µg Lane 4 : MM453 Whole Cell Lysate at 40 µg Lane 5 : SMMC-7721 Whole Cell Lysate at 40 µg
Aquaporin 3 was immunoprecipitated using 0.5mg Mouse Kidney extract, 5µg of Rabbit polyclonal to Aquaporin 3 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-). The antibody was incubated under agitation with Protein G beads for 10min, Mouse Kidney extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation. Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab125219. Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697). Band: 31kDa: Aquaporin 3.
ICC/IF image of ab125219 stained MCF-7 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab125219 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
References for Anti-Aquaporin 3 antibody (ab125219)
This product has been referenced in:
Na TY et al. The trisaccharide raffinose modulates epidermal differentiation through activation of liver X receptor. Sci Rep7:43823 (2017).
Read more (PubMed: 28266648) »
Nazemi S et al. Reciprocity in the developmental regulation of aquaporins 1, 3 and 5 during pregnancy and lactation in the rat. PLoS One9:e106809 (2014).
Read more (PubMed: 25184686) »