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StressMarq/Anti-ENaC Alpha Antibody/SPC-403D-A594/100-µg
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StressMarq/Anti-ENaC Alpha Antibody/SPC-403D-A594/100-µg
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SPC-403D-A594
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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

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

<p>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.</p>

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.

<p>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.</p>

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

 ATTO 594
Overview:

  • High fluorescence yield
  • High photostability
  • Very hydrophilic
  • Excellent solubility in water
  • Very little aggregation
  • New dye with net charge of -1
  • Molar Mass: 1137 g/mol

ATTO 594 Datasheet

 ATTO 594 Fluorophore Excitation and Emission SpectrumOptical Properties:

λex = 601 nm

λem = 627 nm

εmax = 1.2×105

Φf = 0.85

τfl = 3.5 ns

Brightness = 102

Laser = 594 nm

Filter set = Texas Red®

 

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最好有图片说明,谢谢
实验室的一份protocol 上写配上样缓冲液的时候要加溴酚蓝bromophenol blue,但没写加的量是多少。实验室只有粉末状的溴酚蓝。请问大神们如何加,加多少啊?需要先把溴酚蓝配制成溶液吗?还是直接加进去?
上样缓冲液也有多种,一般是80ul的样本加20ul的上样缓冲液,这样基本上上样50ug的体积在10ul左右。
最好是测浓度,一般不超过30-40ug,上样缓冲液一般4×或者5×,这个没太大区别。

实验小白刚刚起步,一般采用等质量上样的方法,但我算的上样量是没加缓冲液之前的,加完缓冲液蛋白样品浓度会变么?师姐说不会变,就按照算好的上样量加就行。

前几天提的样按50ug算每个样本的上样量也只有上5ul左右,按师姐说的就那么上了,结果发完光只有内参有条带,目标蛋白一点也没有,但以前做过一次是出过一点趋势的,而且这次整个过程感觉做的很仔细,发完光结果也很干净整齐,所以我在想是不是跟上样量有关系?上样缓冲液加入之后会使样本浓度稀释五倍么,这样的话上样量就得乘5。

变性前后加上样缓冲液的区别是什么呢?哪一种会改变蛋白浓度从而改变上样量呢?


蛋白上样缓冲液
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

提了蛋白,加上样缓冲液煮过,但发现还是有很多胶状的沉淀,比较黏稠,堵枪头。于是自作主张又超声了一次,结果跑了一次,内参都没有。煮过之后的蛋白不能超声么?

sds电泳上样缓冲液如何配置
2ul 5×上样缓冲液中添加3ul水,混匀,即得2×上样缓冲液
电泳缓冲液的主要作用是使胶的导电性和体系的一致,这样跑出的条带才回一致;
加样缓冲液的主要作用是使PCR产物与其混合,
使DNA沉于加样孔的底部,防止DNA跑出来.
(以下内容仅供参考)
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
4×蛋白上样缓冲液使用说明123
songjunlai19922016-03-22

请问有人知道蛋白上样缓冲液应该怎么配制吗?请赐教