Na k atpase

Manufactured by Merck Group

Sourced in United States

The Na/K ATPase is an enzyme that plays a crucial role in maintaining the electrochemical gradient across the cell membrane. It is responsible for the active transport of sodium and potassium ions, which is essential for various cellular processes, such as regulating cell volume, maintaining membrane potential, and facilitating the uptake of nutrients and the excretion of waste products.

Automatically generated - may contain errors

Lab products found in correlation

β actin, by Abcam (2 mentions) β actin, by Merck Group (2 mentions) Syntaxin, by Santa Cruz Biotechnology (1 mentions) Snap25, by Abcam (1 mentions) Snap23, by Abcam (1 mentions) Vamp2, by Abcam (1 mentions) Mem per plus membrane protein extraction kit, by Thermo Fisher Scientific (1 mentions) Tubulin, by Abcam (1 mentions) Polyclonal anti actin, by Santa Cruz Biotechnology (1 mentions) Polyvinylidene difluoride membrane, by Merck Group (1 mentions) Rab27a, by Abcam (1 mentions) Lamp1, by Merck Group (1 mentions) β enac, by Abcam (1 mentions) γ enac, by Abcam (1 mentions) Gapdh, by Abcam (1 mentions) Peroxidase conjugated secondary antibody, by GE Healthcare (1 mentions) Completetm, by Roche (1 mentions) Alexa fluor 546 conjugated phalloidin, by Thermo Fisher Scientific (1 mentions) Alexa fluor 594 conjugated goat anti mouse igg, by Thermo Fisher Scientific (1 mentions) Alexa fluor 488 conjugated goat anti rabbit igg, by Thermo Fisher Scientific (1 mentions)

18 protocols using na k atpase

Interaction of AgNP with Extracellular and Intracellular Enzymes

Check if the same lab product or an alternative is used in the 5 most similar protocols

Alkaline phosphatase (Sigma-Aldrich) was select as a representative extracellular algal enzyme to study the interaction with AgNP. The effect of AgNP to Alkaline phosphatase was assayed in MOPS by determining enzyme activity, using fluorescently linked 4-methylumbelliferyl phosphate disodium salt as substrate [29 ].
In fish cell exposures, to identify the proteins binding to AgNP in cells, AgNP-protein corona complexes were recovered from intact subcellular compartments isolated by subcellular fractionation and proteins lysed from the AgNP to be detected by mass spectrometry. The identified proteins were analyzed by DAVID (http://david.abcc.ncifcrf.gov/), a protein ontology analysis tool. Among the identified proteins, Na⁺/K⁺-ATPase (Sigma-Aldrich, No. A7510) was selected to study the interaction of AgNP with intracellular proteins. The effect of AgNP on Na⁺/K⁺-ATPase activity was measured in a buffer containing 20 mM Tris–HCl, 0.60 mM EDTA, 5 mM MgCl₂, 3 mM KCl and 133 mM NaCl (pH 7.8) and substrate, ATP (Sigma-Aldrich, No. A9062) [28 (link)].

Yue Y., Li X., Sigg L., Suter M.J., Pillai S., Behra R, & Schirmer K. (2017). Interaction of silver nanoparticles with algae and fish cells: a side by side comparison. Journal of Nanobiotechnology, 15, 16.

+ Open protocol

+ Expand

Mouse Brain Protein Expression Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cell cytosolic or membrane protein lysates of mouse brain tissues were prepared using Mem-PER Plus Membrane Protein Extraction Kit (Thermo Scientific, 89842). Then, the lysates were separated by SDS–PAGE and probed with specific antibodies: SNAP-25 (Abcam, ab66066), SNAP-23 (Abcam, ab3340), syntaxin (Santa Cruz, sc-12736), Vamp2 (Abcam, ab6276), Munc-18 (SYSY, 116002), Phospho-Synaptotagmin (R&D Systems, PPS085), β-ACTIN (Abcam, ab6276), TUBULIN (Abcam, ab15246), and Na/K ATPase (Millipore, 05-369). For quantification, the densitometry measurement of each band was first normalized to that of β-ACTIN, TUBULIN, or Na/K ATPase (used as loading control) and then averaged from at least three independent samples.

Yang H., Zhang M., Shi J., Zhou Y., Wan Z., Wang Y., Wan Y., Li J., Wang Z, & Fei J. (2017). Brain-Specific SNAP-25 Deletion Leads to Elevated Extracellular Glutamate Level and Schizophrenia-Like Behavior in Mice. Neural Plasticity, 2017, 4526417.

+ Open protocol

+ Expand

Antibody Sources for Membrane Protein Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

Antibodies used in the study were from the following sources: monoclonal anti-NHE1 and monoclonal anti-α₁ subunit of the Na/K-ATPase (Millipore, Billerica, MA), polyclonal anti-actin (Santa Cruz Biotechnology, Santa Cruz, CA). Polyclonal rabbit anti-NaPi-IIa, anti-NaPi-IIb, anti-NaPi-IIc, anti-Pit-1 and anti-Pit-2 were characterized and validated previously [16 (link)–18 (link)]. Immunoblotting was performed as described [36 (link)].

Albano G., Moor M., Dolder S., Siegrist M., Wagner C.A., Biber J., Hernando N., Hofstetter W., Bonny O, & Fuster D.G. (2015). Sodium-Dependent Phosphate Transporters in Osteoclast Differentiation and Function. PLoS ONE, 10(4), e0125104.

+ Open protocol

+ Expand

Western Blot Analysis of Protein Expression

Cited 2 times

Check if the same lab product or an alternative is used in the 5 most similar protocols

The protocol was described before [35 (link)–37 (link)]. Equal amounts of proteins (10 μg) or equal volumes of samples (15 μl, for biotinylation of surface protein assay or immunoprecipitation assay) were separated by SDS-PAGE (10% gel) and then transferred to polyvinylidene difluoride membranes (Millipore), membranes were incubated overnight at 4°C, treated with primary antibodies against N-type calcium channels (1: 200, rabbit polyclonal, Millipore), N-type calcium channels (1:500, mouse monoclonal, Santa Cruz), LAMP1 (1:1000, rabbit polyclonal, Sigma), Na⁺/K⁺ ATPase (1:1000, mouse monoclonal, Millipore), and Rab27a (1:1000, rabbit polyclonal, Abcam). β-actin (1: 10,000, mouse monoclonal, Abcam) was used as a loading control. The immunoblots were developed with enhanced chemiluminescence, and bands were visualized and analyzed by LabWorks 4.5 software on a UVP Bioimaging System (Upland). Quantification of results was performed by densitometry. Densitometric intensities of target proteins as well as loading controls (actin, Na/K ATPase or IgG) were quantified with LabWorks software. The optical density ratios of target proteins vs. loading controls were calculated and results were normalized to control values of 1.

Hui L., Geiger N.H., Bloor-Young D., Churchill G.C., Geiger J.D, & Chen X. (2015). Release of calcium from endolysosomes increases calcium influx through N-type calcium channels: Evidence for acidic store-operated calcium entry in neurons. Cell calcium, 58(6), 617-627.

+ Open protocol

+ Expand

Antibody-Based Protein Analysis of Tissue Sections

Check if the same lab product or an alternative is used in the 5 most similar protocols

For western blot and immunofluorescence analyses of tissue sections, we used previously characterized specific antibodies. For western blot, the following antibodies were used: αENaC, βENaC, γENaC (Masilamani et al., 1999 (link)), NHE3 (ab95299, Abcam, Cambridge, United Kingdom), NaPi (LL696AP) (Biber et al., 1996 (link); Kwon et al., 2000 (link)), NKCC2 (LL320AP) (Ecelbarger et al., 1996 (link); Nielsen et al., 1998 (link); Kim et al., 1999 (link)), and Na/K-ATPase (Millipore, 05-369). The following antibodies were used for IHC: βENaC (GTX41971, GeneTeX, Irvine, CA, United States) and γENaC (Masilamani et al., 1999 (link)). Horseradish-conjugated secondary antibodies were used (Life Technologies, Thermo Fisher Scientific, Cambridge, MA, United States). Furthermore, polyclonal goat anti-mouse IgG [P447] or polyclonal goat anti-rabbit IgG [P448] (Dako, Glostrup, Denmark) was used.

Tingskov S.J., Kwon T.H., Frøkiær J, & Nørregaard R. (2018). Tamoxifen Decreases Lithium-Induced Natriuresis in Rats With Nephrogenic Diabetes Insipidus. Frontiers in Physiology, 9, 903.

+ Open protocol

+ Expand

Western Blotting of Ion Channels

Check if the same lab product or an alternative is used in the 5 most similar protocols

Western blotting was performed as described previously (Bomben and Sontheimer, 2008 (link)). Primary antibodies KCa3.1 (1:500) from Sigma, NaKATPase (1:500) from Millipore, and GAPDH (1:5000) from Abcam were used with secondary antibodies conjugated to horseradish peroxidase.

Turner K.L., Honasoge A., Robert S.M., McFerrin M.M, & Sontheimer H. (2014). A pro-invasive role for the Ca2+-activated K+ channel KCa3.1 in malignant glioma. Glia, 62(6), 971-981.

+ Open protocol

+ Expand

Quantitative Protein Analysis in Renal Tissues

Check if the same lab product or an alternative is used in the 5 most similar protocols

For semiquantitative immunoblotting and immunohistochemistry, we used previously characterized polyclonal antibodies. Affinity-purified polyclonal antibodies against NHE3, NKCC2, NCC, ENaC-α, ENaC-γ, NBC, pendrin, and H-ATPase were used as described in a previous study (7 (link), 14 (link)). Affinity-purified polyclonal antibodies against Na-glucose cotransporter-1 (SGLT1; AB1352; Millipore Corp., Billerica, MA, USA), ENaC-β (sc-21013; Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA), and Na-K-ATPase (05-396; Millipore Corp.) were also used. Tissue ET-1 concentrations were measured using QuantiGlo ET-1 ELISA (QET00B; R&D Systems, Minneapolis, MN, USA) according to the manufacturer's instructions.

Kim S., Lee J., Heo N.J., Lee J.W, & Han J.S. (2014). Alkali Therapy Attenuates the Progression of Kidney Injury via Na/H Exchanger Inhibition in 5/6 Nephrectomized Rats. Journal of Korean Medical Science, 29(5), 691-698.

+ Open protocol

+ Expand

CFTR Maturation Quantification in HEK293 Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

After 48 h of transfection, HEK293 cells were lysed (lysis buffer: 10 mM Tris HCl, 1% Non-idet P-40, 0.5% sodium deoxycholate, 1 mM Pefabloc^® SC and the protease inhibitors cocktail cOmplete^TM [Roche, Germany); pH 7.5)]. 50 μg of protein were resolved on a 5–10% gradient SDS-PAGE, transferred to a nitrocellulose membrane, probed using the MAB3480 anti-CFTR antibody (a.a 1370–1380, clone M3A7) and the Na/K ATPase (1:1000; Millipore Corporation, United States), then incubated with a secondary peroxidase-conjugated antibody (1:5000; Amersham, GE Healthcare, United Kingdom) followed by chemiluminescence detection.
Maturation level was quantified by densitometry using ImageJ software (Wayne Rasband, National Institute of Health, United States) and normalized to the loading control, the Na/K ATPase protein.

Billet A., Elbahnsi A., Jollivet-Souchet M., Hoffmann B., Mornon J.P., Callebaut I, & Becq F. (2020). Functional and Pharmacological Characterization of the Rare CFTR Mutation W361R. Frontiers in Pharmacology, 11, 295.

+ Open protocol

+ Expand

Immunofluorescence Staining of HCECs

Check if the same lab product or an alternative is used in the 5 most similar protocols

HCECs cultured on a 48-well cell culture plate were fixed in 0.5% paraformaldehyde for 45 minutes, permeabilized with 1%Triton X-100 for 5 minutes, and incubated with 2% bovine serum albumin (BSA) for 30 minutes. The HCECs were incubated with the following primary antibodies for 45 minutes at 37°C: ZO-1 (1:200, Zymed Laboratories, South San Francisco, CA), N-cadherin (1: 300, BD Biosciences), and Na⁺/K⁺-ATPase (1:200, Merck Millipore). Either Alexa Fluor 488-conjugated goat anti-rabbit IgG (Thermo Fisher Scientific) or Alexa Fluor 594-conjugated goat anti-mouse IgG (Thermo Fisher Scientific) was diluted at 1:500 in BSA, and the samples were incubated in the secondary antibodies for 45 minutes at 37°C. Actin was stained by incubating samples with a 1:200 diluted Alexa Fluor 546-conjugated phalloidin (Life Technologies Corp.) for 45 minutes at 37°C. Nuclei were stained with 1:1000 diluted 4’,6-diamidino-2-phenylindole dihydrochloride (DAPI) (Vector Laboratories, Burlingame, CA). The samples were examined by fluorescence microscopy (BZ-9000; Keyence, Osaka, Japan).

Okumura N., Kagami T., Watanabe K., Kadoya S., Sato M, & Koizumi N. (2019). Feasibility of a cryopreservation of cultured human corneal endothelial cells. PLoS ONE, 14(6), e0218431.

+ Open protocol

+ Expand

Antibody Detection Protocol for Cellular Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols

The basic procedures are the same as described previously.¹⁰ ^,¹⁴ (link)^–¹⁶ (link) The primary antibodies were obtained as follows: Na⁺/K⁺-ATPase (#05-369; Merck KGaA, Darmstadt, Germany); β-actin (#4967), heme oxygenase-1 (HO-1; #5061), p53 (#9282), phospho-p53 (Ser15; #9284), phospho-H2A histone family member X (H2AX, Ser139; #9718), sirtuin 1 (SIRT1; #9475), p21 (#2946), and mitochondrial transcription factor A (TFAM; #8076) from Cell Signaling Technology (Danvers, MA, USA); c-Myc (ab32072), DJ-1 (ab18275), Kelch-like ECH-associated protein 1 (Keap1; ab66620), nuclear factor erythroid 2-related factor 2 (Nrf2; ab62352), translocase of outer mitochondrial membrane 20 (TOMM20; ab56783), and voltage-dependent anion channel (VDAC; ab235143) from Abcam (Cambridge, UK); histone H3 (#05-928; MilliporeSigma, Temecula, CA, USA); PGC1-a (NBP1-04676; Novus Biologicals, Littleton, CO, USA); and Rho A (ab54835) and MMP-2 (ab92536) from Abcam. Goat anti-mouse immunoglobulin G (IgG) and goat anti-rabbit IgG (H+L; Southern Biotech, Birmingham, AL, USA) were used as the secondary antibodies. MagicMark XP Western Protein Standard (Thermo Fisher Scientific, Waltham, MA, USA) was used as the molecular weight marker. The protein bands were made visible by a Western BLoT HRP substrate series (TakaraBio, Shiga, Japan).

Hamuro J., Asada K., Ueno M., Yamashita T., Mukai A., Fujita T., Ito E., Hiramoto N., Toda M., Sotozono C, & Kinoshita S. (2022). Repressed miR-34a Expression Dictates the Cell Fate to Corneal Endothelium Failure. Investigative Ophthalmology & Visual Science, 63(4), 22.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!