Overview:
| Product Name | ENaC Alpha Antibody | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Description | Rabbit Anti-Rat ENaC Alpha Polyclonal | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Species Reactivity | Mouse, Rat, African clawed frog (Xenopus laevis) | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Applications | WB, IHC, ICC/IF | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Antibody Dilution | WB (1:1000), ICC/IF (1:400); optimal dilutions for assays should be determined by the user. | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Host Species | Rabbit | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Immunogen Species | Rat | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Immunogen | Produced against a synthetic peptide mapping to the N-terminal of the alpha subunit (amino acids 46-68) of rat Alpha ENaC (antibody designation 3560-2). | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Concentration | 1 mg/ml | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Conjugates |
Alkaline Phosphatase, APC, ATTO 390, ATTO 488, ATTO 565, ATTO 594, ATTO 633, ATTO 655, ATTO 680, ATTO 700, Biotin, FITC, HRP, PE/ATTO 594, PerCP, RPE, Streptavidin, Unconjugated
StreptavidinProperties:
Streptavidin Datasheet Biotin | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| R-PE (R-Phycoerythrin) | ||
Overview:
R-PE Datasheet |  | Optical Properties: λex = 565 nm λem = 575 nm εmax = 2.0×106 Φf = 0.84 Brightness = 1.68 x 103 Laser = 488 to 561 nm Filter set = TRITC |
Properties
| Storage Buffer | PBS, 50% glycerol, 0.09% sodium azide |
| Storage Temperature | -20ºC |
| Shipping Temperature | Blue Ice or 4ºC |
| Purification | Protein A purified |
| Clonality | Polyclonal |
| Specificity | Detects ~85kDa. |
| Cite This Product | StressMarq Biosciences Cat# SPC-403, RRID: AB_10640131 |
| Certificate of Analysis | 1 µg/ml of SPC-403 was sufficient for detection of alpha-ENaC in 35 µg of rat kidney tissue lysate by colorimetric immunoblot analysis using Goat anti-rabbit IgG:HRP as the secondary antibody. |
Biological Description
| Alternative Names | SCNN1A Antibody, Epithelial Sodium Channel-α Antibody, Epithelial Sodium Channel alpha Antibody, Alpha ENaC 2 Antibody, Alpha ENaC Antibody, Alpha NaCH Antibody, Alpha-ENaC Antibody, Amiloride sensitive epithelial sodium channel alpha subunit Antibody, Amiloride sensitive sodium channel subunit alpha Antibody, Amiloride-sensitive sodium channel subunit alpha Antibody, ENaCa Antibody, ENaCalpha Antibody, Epithelial Na(+) channel subunit alpha Antibody, Epithelial Na+ channel subunit alpha Antibody, FLJ21883 Antibody, Nonvoltage gated sodium channel 1 subunit alpha Antibody, Nonvoltage-gated sodium channel 1 subunit alpha Antibody, SCNEA Antibody, SCNN 1 Antibody, SCNN1 Antibody, SCNN1A Antibody, SCNNA_HUMAN Antibody, Sodium channel nonvoltage gated 1 alpha Antibody |
| Research Areas | Epithelial Sodium Channels (ENaC), Ion Channels, Neuroscience, Sodium Channels |
| Cellular Localization | Apical cell membrane |
| Accession Number | NP_113736 |
| Gene ID | 25122 |
| Swiss Prot | Q6IRJ1 |
| Scientific Background | The Epithelial Sodium Channel (ENaC) is a membrane ion channel permeable to Na+ ions. It is located in the apical plasma membrane of epithelia in the kidneys, lung, colon, and other tissues where it plays a role in trans epithelial Na+-ion transport (1). Specifically Na+ transport via ENaC occurs across many epithelial surfaces, and plays a key role in regulating salt and water absorption (2). ENaCs are composed of three structurally related subunits that form a tetrameric channel, α, β, and γ. The expression of its alpha and beta subunits is enhanced as keratinocytes differentiate (3, 4). The beta and gamma-ENaC subunits are essential for edema fluid to exert its maximal effect on net fluid absorption by distal lung epithelia(5). And it has been concluded that the subunits are differentially expressed in the retina of mice with ocular hypertension, therefore the up-regulation of alpha-ENaC proteins could serve as a protection mechanism against elevated intraocular pressure (6). |
| References |
1. Kakizoe Y., et al. (2009) J Hpyertens. 27(8): 1679-1689. 2. Gu Y. (2008) J Cell Physiol. 216(2):453-457. 3. Bruns J.B. (2003) Am J Physiol Renal Physiol. 285(4): F600-F609. 4. Mauro T., et al. (2002) J Invest Dermatol. 118(4): 589-594. 5. Elias N., et al. (2007) Am J Physiol Lung Cell Mol Physiol. 293(3): L537-45. 6. Dyka F.M., May C.A. and Enz R. (2005) J Neurochem. 94(1): 120-128. |
Product Images
Western blot analysis of Rat kidney tissue lysates showing detection of ENaC protein using Rabbit Anti-ENaC Polyclonal Antibody (SPC-403). Primary Antibody: Rabbit Anti-ENaC Polyclonal Antibody (SPC-403) at 1:1000.
Western blot analysis of Mouse kidney tissue lysates showing detection of ENaC protein using Rabbit Anti-ENaC Polyclonal Antibody (SPC-403). Primary Antibody: Rabbit Anti-ENaC Polyclonal Antibody (SPC-403) at 1:1000.
Product Citations (34)
Western Blot
Enac Regulation in the Kidney: the Role of Ankyrin G
Klemens, C.A. (2017) University of Pittsburgh. PhD Dissertation.
PubMed ID: N/A Reactivity Rat Applications: Western Blot
Derlin-1 promotes ubiquitination and degradation of epithelial sodium channel (ENaC).
You, H. et al. (2017) J Cell Sci. 130(6):1027-1036.
PubMed ID: 28137758 Reactivity Mouse Applications: Western Blot
Derlin-1 promotes ubiquitination and degradation of epithelial sodium channel (ENaC).
You, H. et al. (2017) J Cell Sci. 130(6):1027-1036.
PubMed ID: 28137758 Reactivity Human Applications: Western Blot
Ankyrin G Expression Regulates Apical Delivery of the Epithelial Sodium Channel (ENaC).
Klemens, C.A., Edinger, R.S., Kightlinger, L., Liu, X. and Butterworth, M.B. (2016) J Biol Chem. 292(1):375-385.
PubMed ID: 27895120 Reactivity Mouse Applications: Western Blot
Renal sodium transport in renin-deficient Dahl salt-sensitive rats.
Pavlov, T.S., Levchenko, V., Ilatovskaya, D.V., Moreno, C. and Staruschenko, A. (2016) J Renin Angiotensin Aldosterone Syst. 17(3).
PubMed ID: 27443990 Reactivity Rat Applications: Western Blot
(Pro)Renin Receptor Regulates Potassium Homeostasis through a Local Mechanism.
Xu, C. et al. (2016) Am J Physiol Renal Physiol. [Epub ahead of print]
PubMed ID: 27440776 Reactivity Rat Applications: Western Blot
AMP-activated protein kinase attenuates high salt-induced activation of epithelial sodium channels (ENaC) in human umbilical vein endothelial cells.
Zheng, W. et al. (2016) Oxid Med Cell Longev. 2016:1531392.
PubMed ID: 27635187 Reactivity Human Applications: Western Blot
Renal tubular epithelial cell prorenin receptor regulates blood pressure and sodium transport.
Ramkumar, N. et al. (2016) Am J Physiol Renal Physiol. 311(1):F186-94.
PubMed ID: 27053687 Reactivity Mouse Applications: Western Blot
The Role of Epithelial Sodium Channel ENaC and the Apical Cl-/HCO3- Exchanger Pendrin in Compensatory Salt Reabsorption in the Setting of Na-Cl Cotransporter (NCC) Inactivation.
Patel-Chamberlin, M. et al. (2016) PLoS One. 11(3):e0150918.
PubMed ID: 26963391 Reactivity Mouse Applications: Western Blot
Possible role for nephron-derived angiotensinogen in angiotensin-II dependent hypertension.
Ramkumar, N. et al. (2016) Physiol Rep. 4(1). pii: e12675.
PubMed ID: 26755736 Reactivity Mouse Applications: Western Blot
The Thiazide-Sensitive Co-Transporter Promotes the Development of Sodium Retention in Mice with Diet-Induced Obesity.
Davies, M. R. P. et al. (2015) Kidney Blood Press Res. 40(5):509-519.
PubMed ID: 26418861 Reactivity Mouse Applications: Western Blot
Inhibition of Mitochondrial Complex-1 Prevents the Downregulation of NKCC2 and ENaCα in Obstructive Kidney Disease.
Zhang, Y. et al. (2015) Sci Rep. 5:12480.
PubMed ID: 26207612 Reactivity Mouse Applications: Western Blot
Increased Epithelial Sodium Channel Activity Contributes to Hypertension Caused by Na+-HCO3− Cotransporter Electrogenic 2 Deficiency.
Wen, D. et al. (2015) Hypertension. 66(1):68-74.
PubMed ID: 25941340 Reactivity Mouse Applications: Western Blot
Compositions And Methods Of Use For (PRO)Renin Receptor.
Feng, Y. and Yang, T. (2015) United States Patent Application 20150025013.
PubMed ID: N/A Reactivity Rat Applications: Western Blot
Role of Rho GDP-dissociation inhibitor alpha in control of ENaC-mediated sodium reabsorption.
Pavlov, T. S., Levchenko, V. and Staruschenko, A. (2014) J Biol Chem. 289(41):28651-9.
PubMed ID: 25164814 Reactivity Mouse Applications: Western Blot
Collecting duct specific knock-out of renin attenuates angiotensin-II induced hypertension.
Ramkumar, N. et al. (2014) Am J Physiol Renal Physiol. 307(8):F931-8.
PubMed ID: 25122048 Reactivity Mouse Applications: Western Blot
Novel Mechanisms of Sodium Retention in Obesity: Phosphorylation of NKCC2 and Regulation of SPAK/OSR1 by AMPK.
Davies, M. R. P. et al. (2014) Am J Physiol. 307(1):F96-F106.
PubMed ID: 24808538 Reactivity Mouse Applications: Western Blot
Chronic Angiotensin II Infusion Drives Extensive Aldosterone-Independent Epithelial Na+ Channel Activation.
Mamenko,M. et. al. (2013) Hypertension. 62(6):1111-22.
PubMed ID: 24060890 Reactivity Mouse Applications: Western Blot
Adenylyl Cyclase VI Mediates Vasopressin-Stimulated ENaC Activity.
Roos, K.P. et al. (2012) JASN. 24 (2): 218-227.
PubMed ID: 23264685 Reactivity Mouse Applications: Western Blot
Increased expression of renal TRPM6 compensates for Mg2+ wasting during furosemide treatment.
van Angelen, A.A., Kemp, A.W., Hoenderop, J.G. and Bindels, B.J. (2012) Clin Kidney J. 5 (6): 535-544.
PubMed ID: 26069797 Reactivity Mouse Applications: Western Blot
Regional differences in rat conjunctival ion transport activities.
Yu, D. et al. (2012) Am J Physiol Cell Physiol. 303 (7): 767-780.
PubMed ID: 22814399 Reactivity Xenopus laevis Applications: Western Blot
The Epithelial Sodium Channel (ENaC) Establishes a Trafficking Vesicle Pool Responsible for Its Regulation.
Edinger, R.S. et al. (2012) PLoS ONE. 7(9): e46593.
PubMed ID: 23029554 Reactivity Mouse Applications: Western Blot
Aldosterone does not require angiotensin II to activate NCC through a WNK4–SPAK–dependent pathway.
van der Lubbe, N. et al. (2012) Euro J Physiol. 463 (6): 853-863.
PubMed ID: 22549242 Reactivity Rat Applications: Western Blot
Immunohistochemistry
The Prostaglandin E2 Receptor 1 (EP1) Antagonizes AngII in the Collecting Duct.
Eckert, D. (2017) University of Ottawa. Dissertation.
PubMed ID: N/A Reactivity Mouse Applications: Immunohistochemistry
Inhibition of Mitochondrial Complex-1 Prevents the Downregulation of NKCC2 and ENaCα in Obstructive Kidney Disease.
Zhang, Y. et al. (2015) Sci Rep. 5:12480.
PubMed ID: 26207612 Reactivity Mouse Applications: Immunohistochemistry
Epoxyeicosatrienoic Acid (EET) Analog Lowers Blood Pressure Through Vasodilation And Sodium Channel Inhibition.
Khan, M. A., et al. (2014) Clin Sci (Lond). 127(7):463-74.
PubMed ID: 24707975 Reactivity Rat Applications: Immunohistochemistry
Chronic Angiotensin II Infusion Drives Extensive Aldosterone-Independent Epithelial Na+ Channel Activation.
Mamenko,M. et. al. (2013) Hypertension. 62(6):1111-22.
PubMed ID: 24060890 Reactivity Mouse Applications: Immunohistochemistry
ENaC γ-expressing astrocytes in the circumventricular organs, white matter, and ventral medullary surface: sites for Na+ regulation by glial cells.
Miller, R.L. and Loewy, A.D. (2013) J Chem Neuroanat. 53:72-80.
PubMed ID: 24145067 Reactivity Rat Applications: Immunohistochemistry
Regulation of ENaC in mice lacking renal insulin receptors in the collecting duct.
Pavlov, T.S. et. al. (2013) FASEB J. 27(7):2723-32.
PubMed ID: 23558339 Reactivity Mouse Applications: Immunohistochemistry
Other Citations
Derlin-1 promotes ubiquitination and degradation of epithelial sodium channel (ENaC).
You, H. et al. (2017) J Cell Sci. 130(6):1027-1036.
PubMed ID: 28137758 Reactivity Mouse Applications: Immunoprecipitation
Derlin-1 promotes ubiquitination and degradation of epithelial sodium channel (ENaC).
You, H. et al. (2017) J Cell Sci. 130(6):1027-1036.
PubMed ID: 28137758 Reactivity Human Applications: Immunoprecipitation
ENaC and ROMK activity are inhibited in the DCT2/CNT of TgWnk4PHAII mice.
Zhang, C. et al. (2016) Am J Physiol Renal Physiol. [Epub ahead of print]
PubMed ID: N/A
mTORC1 maintains renal tubular homeostasis and is essential in response to ischemic stress.
Grahammer, F. et al. (2014) Proc Natl Acad Sci U S A. 111(27):E2817-26.
PubMed ID: 24958889 Reactivity Mouse Applications: Gel Shift
mPGES-1-Derived PGE2 Mediates Dehydration Natriuresis.
Jia, Z. et al. (2013) Am J Physiol Renal Physiol. 304(2):F214-21.
PubMed ID: 23171554 Reactivity Mouse Applications: Western Blot
Streptavidin
Properties:
- Homo-tetrameric protein purified from Streptomyces avidinii which binds four biotin molecules with extremely high affinity
- Molecular weight: 53 kDa
- Formula: C10H16N2O3S
- Applications: Western blot, immunohistochemistry, and ELISA
Streptavidin Datasheet
ebiomall.com
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① 上样缓冲液中包括甘油,溴酚蓝,SDS等,甘油主要作用是防止样品漂浮出点样孔,溴酚蓝作为电泳指示剂,用来观察电泳进度,SDS用于一些核酸结合蛋白的变性解离.
②核酸染料与凝胶中核酸结合,以便于在紫外光下显示核酸条带.
蛋白电泳上样缓冲液包括2%SDS , 0.1%溴酚蓝 10%甘油,SDS是保证样品中的所有蛋白带电荷一致，减少电荷对电泳结果的影响。还原剂是让二硫键处于断开状态，保证蛋白分子的线性.
溴酚蓝作用是指示上样蛋白的跑胶时标记的，甘油是增加上样重量的，以免漂浮，煮沸是使蛋白变性的永久保存
2乘SDS-PAGE上样缓冲液（20ml体系）
1.0mol/L Tris-HCL (pH 6.8) 2ml
1.0mol/L DTT 4ml
SDS 0.8g
溴芬蓝 0.04g
甘油 4ml
dd水 定容至20ml
4℃冰箱保存，可以不分装，我们实验室一般用三个月以上
先配1.0mol/L的DTT,双蒸无菌水溶解4℃冰箱保存,配上样缓冲液的时候加入就可以了,都混匀保存也没问题的
1mol/l Tirs-HCl(PH8.8) 0.6ml
10%SDS 2ml
50%甘油 5ml
2-巯基乙醇 0.5ml
1%溴酚兰 1ml
水 0.9ml
另外：SDS不好溶解建议加热溶解,如果要做非还原SDS-PAGE请将2-巯基乙醇 0.5ml换成超纯水即可
电泳缓冲液的另一个作用是使溶液具有一定的导电性，以利于DNA分子的迁移，例如，一般电泳缓冲液中应含有0.01-0.04mol/L的Na+离子，Na+离子的浓度太低时电泳速度变慢；太高时就会造成过大的电流使胶发热甚至熔化。
电泳缓冲液还有一个组分是EDTA，加入浓度为1-2mmol/L，目的是螯合Mg2+ 等离子，防止电泳时激活 DNA酶，此外还可防止Mg2+离子与核酸生成沉淀。
因为DNA和RNA链上的碱基都基本一致。而这两种上样缓冲液中含有的染料是二甲苯青和溴酚蓝，都可以与其碱基结合，跑出条带。
但是，如果使用DNA上样缓冲液，里面会夹杂一些RNA酶，这些酶有可能会影响到实验结果。使用前需要将RNA酶处理掉。
