Immunofluorescence analysis of alpha ENaC was performed using 70% confluent log phase HEK 293 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with alpha ENaC Rabbit Polyclonal Antibody (Product # PA1-920A) at 2µg/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjµgate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human, Mouse, Rat|
|Published species reactivity||Rat, Mouse, Human, Not Applicable|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to residues L(20) M K G N K R E E Q G L G P E P A A P Q Q P T(42) C of Human alpha-Enac.|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|Western Blot (WB)||1:100-1:1000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
PA1-920A detects alpha-epithelial sodium channel (alpha-ENaC) from human, mouse, and rat tissues and cells.
PA1-920A has been successfully used in Western blot procedures. By Western blot, this antibody specifically detects a ~97 kDa band representing glycosylated alpha-ENaC from NIH-3T3 cells, and a ~75 kDa band representing the unglycosylated alpha-ENaC protein from human brain samples.
The PA1-920A immunizing peptide is a synthetic peptide corresponding to residues 20-42 from human alpha-ENaC. This sequence is 82%, 73%, 64%, and 59% conserved in the rat and rabbit, mouse, guinea pig, and cow, respectively. PA1-920A immunizing peptide (Cat. # PEP-088) is available for use in neutralization and control experiments.
Epithelial sodium channels are amiloride-sensitive members of the degenerin/epithelial sodium channel (Deg/ENaC) superfamily of ion channels. Members of this superfamily of ion channels share organizational similarity in that they all possess two short intracellular amino and carboxyl termini, two short membrane spanning segments, and a large extracellular loop with a conserved cysteine-rich region. There are three homologous isoforms of the ENaC (alpha, beta, and gamma) protein. ENaC in the kidney, lung, and colon plays an essential role in trans-epithelial sodium and fluid balance. ENaC also mediates aldosterone-dependent sodium reabsorption in the distal nephron of the kidney, thus regulating blood pressure. ENaC is thought to be regulated, in part, through association with the cystic fibrosis transmembrane conductance regulator (CFTR) chloride ion channel. Gain-of-function mutations in beta- or gamma-ENaC can cause severe arterial hypertension (Liddel and quote;s syndrome) and loss-of-function mutations in alpha- or beta-ENaC causes pseudohypoaldosteronism (PHA-1).
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Regulation of mechanosensitive biliary epithelial transport by the epithelial Na(+) channel.
PA1-920A was used in immunohistochemistry to study the epithelial Na(+) channel that transports and regulated mechanosensitive biliary epithelial transport
|Li Q,Kresge C,Bugde A,Lamphere M,Park JY,Feranchak AP||Hepatology (Baltimore, Md.) (63:538)||2016|
Filamin interacts with epithelial sodium channel and inhibits its channel function.
PA1-920A was used in immunohistochemistry to study the inhibition of epithelial sodium channel function by the direct binding of filamin
|Wang Q,Dai XQ,Li Q,Tuli J,Liang G,Li SS,Chen XZ||The Journal of biological chemistry (288:264)||2013|
Endothelial Gata5 transcription factor regulates blood pressure.
PA1-920A was used in western blot to test if GATA5 is a regulator of blood pressure
|Messaoudi S,He Y,Gutsol A,Wight A,Hébert RL,Vilmundarson RO,Makrigiannis AP,Chalmers J,Hamet P,Tremblay J,McPherson R,Stewart AF,Touyz RM,Nemer M||Nature communications (6:null)||2015|
Renal tubule angiotensin II type 1 receptor-associated protein promotes natriuresis and inhibits salt-sensitive blood pressure elevation.
PA1-920A was used in western blot to study the effects of ATRAP in the renal distal tubules in response to high salt treatment using wild type and ATRAP transgenic mice
|Wakui H,Uneda K,Tamura K,Ohsawa M,Azushima K,Kobayashi R,Ohki K,Dejima T,Kanaoka T,Tsurumi-Ikeya Y,Matsuda M,Haruhara K,Nishiyama A,Yabana M,Fujikawa T,Yamashita A,Umemura S||Journal of the American Heart Association (4:null)||2015|
Deletion of the angiotensin II type 1 receptor-associated protein enhances renal sodium reabsorption and exacerbates angiotensin II-mediated hypertension.
PA1-920A was used in western blot to study the effects of ATRAP deletion on renal sodium balance and hypertension mediated by angiotensin II
|Ohsawa M,Tamura K,Wakui H,Maeda A,Dejima T,Kanaoka T,Azushima K,Uneda K,Tsurumi-Ikeya Y,Kobayashi R,Matsuda M,Uchida S,Toya Y,Kobori H,Nishiyama A,Yamashita A,Ishikawa Y,Umemura S||Kidney international (86:570)||2014|
Cholera toxin enhances Na(+) absorption across MCF10A human mammary epithelia.
PA1-920A was used in western blot to study the role of ENaC in the mechanism by which cholera toxin mediates increased human mammary epithelial sodium uptake
|Wang Q,Schultz BD||American journal of physiology. Cell physiology (306:C471)||2014|
Enhancement of alveolar epithelial sodium channel activity with decreased cystic fibrosis transmembrane conductance regulator expression in mouse lung.
PA1-920A was used in western blot to study the inverse relationship between alveolar epithelial sodium channel activity and cystic fibrosis transmembrane conductance regulator expression in mice
|Lazrak A,Jurkuvenaite A,Chen L,Keeling KM,Collawn JF,Bedwell DM,Matalon S||American journal of physiology. Lung cellular and molecular physiology (301:L557)||2011|
Regulation of endogenous ENaC functional expression by CFTR and ¿F508-CFTR in airway epithelial cells.
PA1-920A was used in western blot to investigate the role of CFTR in the modulation of epithelial sodium channel expression and function
|Rubenstein RC,Lockwood SR,Lide E,Bauer R,Suaud L,Grumbach Y||American journal of physiology. Lung cellular and molecular physiology (300:L88)||2011|
Interleukin-6 stimulates epithelial sodium channels in mouse cortical collecting duct cells.
PA1-920A was used in western blot to investigate the effect of interleukin-6 on epithelial sodium channel function in mouse cortical collecting duct cells
|Li K,Guo D,Zhu H,Hering-Smith KS,Hamm LL,Ouyang J,Dong Y||American journal of physiology. Regulatory, integrative and comparative physiology (299:R590)||2010|
Long-term terbutaline exposure stimulates alpha1-Na+-K+-ATPase expression at posttranscriptional level in rat fetal distal lung epithelial cells.
PA1-920A was used in western blot to determine the effect of long-term terbutaline treatment on steady-state expression levels of ENaC in rat fetal distal lung epithelial cells
|Rahman MS,Gandhi S,Otulakowski G,Duan W,Sarangapani A,O'Brodovich H||American journal of physiology. Lung cellular and molecular physiology (298:L96)||2010|
The carboxyl terminus of the alpha-subunit of the amiloride-sensitive epithelial sodium channel binds to F-actin.
PEP-088 was used in western blot to study the interaction of amiloride-sensitive epithelial sodium channel and F-actin
|Mazzochi C,Bubien JK,Smith PR,Benos DJ||The Journal of biological chemistry (281:6528)||2006|