The following primary antibodies were applied for immunofluorescence, immunohistochemistry, or immunoblotting: anti-AQP1 (Alpha diagnostic international, San Antonio, Texas, USA), anti-AQP2 (Santa Cruz Biotechnology, Heidelberg, Germany), anti-phospho-aquaporin 2 (pS256)55 , anti-β-actin (Sigma-Aldrich, St. Louis, USA), anti-Cav1 (Santa Cruz Biotechnology, Heidelberg, Germany), anti-NKCC1 (T4; Developmental Studies Hybridoma Bank, University of Iowa, USA), anti-vasopressin V2 receptor56 (link), anti-eNOS (Santa Cruz Biotechnology), anti-Na+/K+-ATPase (Millipore, Darmstadt, Germany), anti-NCC, anti-NKCC2, anti-phospho-NKCC2 (pT96/pT101), and anti-phospho-NCC (pS71)57 (link).
Anti na k atpase
Manufactured by Merck Group
Sourced in United States
The Anti-Na/K ATPase is a laboratory equipment used to measure the activity of the sodium-potassium ATPase enzyme. The sodium-potassium ATPase is an essential membrane protein responsible for maintaining the electrochemical gradients of sodium and potassium ions across the cell membrane. The Anti-Na/K ATPase device allows researchers to quantify the functional activity of this enzyme, which is crucial for various physiological processes.
Automatically generated - may contain errors
Lab products found in correlation
Anti β actin, by Merck Group (3 mentions)
Anti nkcc1, by Developmental Studies Hybridoma Bank (1 mentions)
Anti aqp2, by Santa Cruz Biotechnology (1 mentions)
Anti cav 1, by Santa Cruz Biotechnology (1 mentions)
Anti enos, by Santa Cruz Biotechnology (1 mentions)
Anti ncc, by Merck Group (1 mentions)
Anti aqp1, by Alpha Diagnostic (1 mentions)
Ab37168, by Abcam (1 mentions)
Ab125066, by Abcam (1 mentions)
Protein block serum free solution, by Agilent Technologies (1 mentions)
Anti fibronectin, by Merck Group (1 mentions)
Anti zo 1, by Thermo Fisher Scientific (1 mentions)
Anti cd73, by BD (1 mentions)
Anti vimentin, by Santa Cruz Biotechnology (1 mentions)
Alexa 488, by Thermo Fisher Scientific (1 mentions)
Goat serum, by Merck Group (1 mentions)
Anti brdu, by Merck Group (1 mentions)
Anti gfp, by Merck Group (1 mentions)
Anti vimentin, by Merck Group (1 mentions)
Microscope dm 2500, by Leica (1 mentions)
13 protocols using anti na k atpase
1
Immunostaining of Aquaporin Proteins
2
Immunohistochemistry of Brain Sections
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Preparation brain sections was as previously described [[30] , [31] ]. Primary antibodies were diluted in 1 % bovine serum albumin (BSA) solution, applied overnight at 4 °C, and washed three times with PBST (PBS solution containing 0.2 % Tween-20). Then, the sections were incubated with secondary antibodies for 3 h at room temperature. After washing three times with PBST, the nuclei were stained utilizing 4',6-diamidino-2-phenylindole (DAPI) for 15 min. We used a laser scanning confocal microscope to acquire images. The primary antibodies used were as follows: anti-FSP1 (39498, SAB) and anti-Na+/K+-ATPase (05–369, Millipore).
3
Corneal Cell Marker Localization
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Part of the corneas was frozen and embedded in OCT compound. Part of the corneas were fixed in 4% formaldehyde and embedded in paraffin. Sections were cut at 5 µm slices, and subjected to immunofluorescence, immunohistochemistry and HE staining according to standard methods. For immunofluorescence and immunohistochemistry, the primary antibody was anti-Na+/K+ ATPase (1:100, Millipore), anti-Pitx2 (1:100, Novus) and anti-Col8a2 (1:50, Novus).
4
Evaluating Ferroptosis Regulators by Western Blotting
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Western blotting was performed as previously described. The following primary antibodies were used: anti-β-actin (A5441, Sigma Aldrich), anti-GAPDH (ab37168, Abcam), anti-glutathione peroxidase 4 (GPX4) (ab125066, Abcam), anti-ferroptosis suppressor protein 1 (FSP1) (39498, SAB), anti-Na+/K+-ATPase (05–369, Millipore), and anti-NMT2 (ABclonal, A7042).
5
Characterizing Corneal Endothelial Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cell morphology was evaluated by using an inverted light microscope (Olympus IX-71, Tokoyo, Japan). The CECs were further characterized for function-related markers, including Na+/K+-ATPase (a pump function associated protein [12] ), and zona occludens-1 (ZO-1, a tight junction associated protein [13] ). Changes of the EnMT were also evaluated by investigating CD73 (a marker to determine the fibroblastic phenotype of CEC [14] ), and fibronectin [15, 16] and vimentin [17, 18] (two mesenchymal markers). Briefly, the cultured cells were first fixed with cold methanol for 20 min. After blocking with Protein Block Serum-Free solution (DAKO, Carpinteria, CA, USA), the cultured cells were incubated overnight with the following primary antibodies: anti-Na+/K+-ATPase (1:300 dilution, Millipore), anti-ZO-1 (1:150 dilution, Invitrogen), anti-CD73 (1:300 dilution; BD Pharmingen Stain Buffer), anti-fibronectin (1:200 dilution, Sigma-Aldrich), and anti-vimentin (1:100 dilution; Santa Cruz). Fluorescein isothiocyanate (FITC) conjugated antibodies (1:200 dilution, Vector Laboratories, Burlingame, CA) were used as the secondary antibodies.
6
Zebrafish Kidney Injury Immunostaining
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Zebrafish were selected 3–6 hours after kidney injury. Then, they were fixed in Dent’s solution (20% DMSO, 80% Methanol) for 24 h at 4°C. They were rehydrated in 75∶25 MeOH/PBSDT, 50∶50, 25∶75 and 0∶100, followed by overnight blocking at 4°C in 10% Goat serum (Sigma) in PBSDT: 1%DMSO, 0.05% Tween20 in 1xPBS. We used the following primary antibodies: anti BrdU (Sigma), anti-GFP (Sigma), anti-Acetilated Tubulin (Sigma), anti-Crumbs (from Dr. Jarema Malicki [34] ), anti-Na/K ATPase (DSHB), anti-Vimentin (Sigma). Anti-mouse and anti-rabbit secondary antibodies were used (Alexa 488 or Alexa 546 labeled, Molecular Probes). All antibody incubations were performed in 2% Goat serum/PBSDT at 4°C overnight. All washes were done at room temperature in 2% Goat serum/PBSDT. Imaging of antibody labeling was performed using Zeiss LSM5 or Nikon C2 confocal microscope.
7
Kidney Immunostaining and Imaging Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Kidney sections and fixed cells were immunostained as described previously [10 (link)]. For immunochemical staining, sections were stained with anti-AQP2 (Cat. No. AQP-002; Alomone Laboratories) antibody. Hematoxylin was used for counter-staining. For immunofluorescence, the following antibodies were used: anti–acetylated (ac)-α-tubulin (Cat. No. T7451; Sigma-Aldrich), anti–Na/K-ATPase (Cat. No. ab76020; Abcam), anti-AQP1 (Cat. No. AQP-001; Alomone Laboratories), and anti-AQP2 (Cat. No. AQP-002). To detect cell nuclei, DAPI was applied to samples. Images were captured using a Leica microscope (DM2500; Wetzlar).
8
Western Blotting Optimization Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Unless otherwise stated whole cell extracts were prepared by lysing cells directly in SDS sample buffer. SDS-PAGE was performed on 12.5% acrylamide gels. For Western blotting 0.2 μm nitrocellulose membranes were used. For blocking, the membranes were incubated in 5% fat-free milk powder and 0.1% Tween-20 in PBS. Antibodies and their sources were: anti-ubiquitin (Z0458, DAKO), anti-Rpt6 (p45-110, Enzo Biosciences), anti-20S proteasome α-subunits (MCP231, Enzo Biosciences), anti-FLCN (D14G9, Cell Signaling Technology), anti-GFP (3H9, Chromotek), anti-myc (9E1, Chromotek) anti-HA (12CA5, Roche), anti-Flag (M2, Sigma), anti-RGSHis (34650, Qiagen), anti-β-actin (AC74, Sigma), anti-NaK-ATPase (C464.6, Sigma), anti-LC3A/B (D3U4C, Cell Signaling Technology), anti-GAPDH (14C10, Cell Signaling Technology), anti-α tubulin (TAT-1, 00020911 Sigma). All secondary antibodies were purchased from DAKO. The Un-Scan-It v6.1 software (Silk Scientific) was used for densitometry.
9
Preparation and Detection of Soluble Membrane Fractions
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Whole cell lysates were prepared from each cancer cell line using the lysis buffer or the Tris/Sucrose/EDTA buffer. The soluble membrane fractions were prepared from the whole cell lysates. Briefly, after centrifugation at 1,000 × g for 10 min, the supernatant was ultracentrifuged at 100,000 × g for 45 min. Then, the soluble membrane fractions were collected by dissolving the precipitates with the Tris/Sucrose/EDTA buffer supplemented with 0.8% (v/v) Nonidet P-40, 0.4% deoxycholic acid, and 0.08% sodium dodecyl sulfate (SDS), followed by a second ultracentrifugation under the same conditions. Using the same procedure, the soluble membrane fraction of human liver tissue was prepared.
The proteins were separated by SDS-polyacrylamide gel electrophoresis, and then transferred onto a polyvinylidene difluoride membrane. The membrane was blocked with 5% skim milk for one hour at room temperature. Affinity purified rabbit polyclonal anti-OATP1B3 antibodies, which recognize the C-terminal region of Lt-OATP1B3, were used as the primary antibodies (1,000-fold dilution, HPA004943, Sigma, St. Louis, MO). Anti-Na+/K+ ATPase (1,000-fold dilution, Sigma) and anti-β-actin (500-fold dilution, Sigma) were also used. Immunocomplexes were detected with ECL Western blotting detection reagents (GE Healthcare, Giles, UK).
The proteins were separated by SDS-polyacrylamide gel electrophoresis, and then transferred onto a polyvinylidene difluoride membrane. The membrane was blocked with 5% skim milk for one hour at room temperature. Affinity purified rabbit polyclonal anti-OATP1B3 antibodies, which recognize the C-terminal region of Lt-OATP1B3, were used as the primary antibodies (1,000-fold dilution, HPA004943, Sigma, St. Louis, MO). Anti-Na+/K+ ATPase (1,000-fold dilution, Sigma) and anti-β-actin (500-fold dilution, Sigma) were also used. Immunocomplexes were detected with ECL Western blotting detection reagents (GE Healthcare, Giles, UK).
10
Immunoblotting for Proximal Tubule Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were collected in RIPA buffer (25 mM Tris HCl pH 7.6, 150 mM NaCl, 1% NP-40, 1% sodium deoxycholate, 0.1% SDS). 30 µg of protein for each sample was loaded on a 10% SDSpolyacrylamide gel, transferred to a PVDF membrane and probed with the following antibodies: a homemade anti-TRPV4 antibody described earlier (Gevaert et al., 2007) , an anti-aquaporin 1 antibody (Sigma) as positive marker of proximal tubule, an anti-Aquaporin 2 antibody (Sigma) as negative marker of PT (Janas et al., 2016) , an anti-megalin antibody (homemade) as apical side marker of PT, an anti-Na + /K + -ATPase (Sigma) as a basolateral side marker of PT and an anti-GAPDH antibody as loading control (Cell Signaling).
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?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!