Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Storage bufferpH: 7.40
Preservative: 0.1% Sodium azide
Constituents: 1% BSA, 0.0268% PBS
Concentration information loading...
PurityProtein G purified
Our Abpromise guarantee covers the use of ab53212 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||Use a concentration of 2 µg/ml. PubMed: 19819874|
|IHC-Fr||Use at an assay dependent concentration. PubMed: 21084687|
FunctionActs as a calcium-activated chloride channel. Required for normal tracheal development.
Tissue specificityBroadly expressed with higher levels in liver and skeletal muscle.
Sequence similaritiesBelongs to the anoctamin family.
DomainThe region spanning the fifth and sixth transmembrane domains probably forms the pore-forming region.
Cellular localizationCell membrane. Cytoplasm.
- Information by UniProt
- ANO 1 antibody
- ANO1 antibody
- ANO1_HUMAN antibody
Immunocytochemistry/ Immunofluorescence analysis of Wt1cre-YFP mouse interstitial cell of Cajal labelling TMEM16A with ab53212 at 1/20 dilution. Cells from the muscular layer, especially the circular one, show TMEM16A (ANO1 - red) immunoreactivity. This immunostaining becomes strong by E16.5. Colocalization of YFP with ANO1 is observed in many cells of both, the circular and the longitudinal layer.
ab53212 (2µg/ml) staining TMEM16A in human liver (left panel) using an automated system (DAKO Autostainer Plus). Using this protocol there is strong staining of hepatocyte cell membrane.
Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer EDTA pH 9.0 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.
Analysis of TMEM16A expression was performed in frozen liver sections (10 µm) from normal and 7-day bile duct-ligated rats and mice.
Sections were defrosted at room temperature and fixed in 4% paraformaldehyde (1× PBS), permeabilized in PBST (1× PBS with 0.2% Triton), and blocked in 4% BSA (in PBST) for 1 hour at room temperature. Samples were then incubated with ab53212 at a 1/100 dilution, diluted in 1% BSA. Sections were incubated overnight at 4 °C and washed three times for 10 minutes each with 1× PBST at room temperature. Sections were incubated with Dylight 488 conjugated donkey anti-rabbit secondary at a 1/600 dilution for 2 hours at room temperature protected from light. Following incubation, the slides were washed in PBST at room temperature and coverslipped with Antifade gold and DAPI. Images were visualized using an Olympus IX-71 confocal microscope.
Scale bar, 10 µm.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human gastrointestinal stromal tumor labeling TMEM16A with ab53212.
ICC/IF image of ab53212 stained HepG2 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 (ab53212, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit 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.
This product has been referenced in:
- Wang P et al. Inflammatory mediators mediate airway smooth muscle contraction through a G protein-coupled receptor-transmembrane protein 16A-voltage-dependent Ca2+ channel axis and contribute to bronchial hyperresponsiveness in asthma. J Allergy Clin Immunol 141:1259-1268.e11 (2018). WB, ICC/IF . Read more (PubMed: 28754608) »
- Song Y et al. Inhibition of ANO1/TMEM16A induces apoptosis in human prostate carcinoma cells by activating TNF-a signaling. Cell Death Dis 9:703 (2018). Read more (PubMed: 29899325) »