Structured Review

Millipore eipa
Effects of <t>DIDS</t> (400 μM) ( A ), AZ (200 μM) ( B ), <t>EIPA</t> (400 μM) ( C ), and bumetanide (Bumex; 400 μM) and/or DIDS (400 μM) ( D ) on Cl − flux at ionocytes of larval skin. Data are presented as the means ± SEM. * p
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Images

1) Product Images from "Salt secretion is linked to acid-base regulation of ionocytes in seawater-acclimated medaka: new insights into the salt-secreting mechanism"

Article Title: Salt secretion is linked to acid-base regulation of ionocytes in seawater-acclimated medaka: new insights into the salt-secreting mechanism

Journal: Scientific Reports

doi: 10.1038/srep31433

Effects of DIDS (400 μM) ( A ), AZ (200 μM) ( B ), EIPA (400 μM) ( C ), and bumetanide (Bumex; 400 μM) and/or DIDS (400 μM) ( D ) on Cl − flux at ionocytes of larval skin. Data are presented as the means ± SEM. * p
Figure Legend Snippet: Effects of DIDS (400 μM) ( A ), AZ (200 μM) ( B ), EIPA (400 μM) ( C ), and bumetanide (Bumex; 400 μM) and/or DIDS (400 μM) ( D ) on Cl − flux at ionocytes of larval skin. Data are presented as the means ± SEM. * p

Techniques Used:

2) Product Images from "Exploring the role of stromal osmoregulation in cancer and disease using executable modelling"

Article Title: Exploring the role of stromal osmoregulation in cancer and disease using executable modelling

Journal: Nature Communications

doi: 10.1038/s41467-018-05414-y

Application of the qualitative network to murine embryonic fibroblasts. Expansion of the QN to p53 −/− , Kras G12D/+ (HET) and p53 −/− , Kras G12D/G12D (HOM) MEFs. a Transport proteins deregulated within HOM MEFs when compared to HET MEFs. Proteins are upregulated (red arrows), downregulated (blue arrows), or unchanged (grey). b Phenotype change for MEFS when HOM MEFs are compared to HET MEFs. The model output represents the physiological behaviour predicted by the model when proteins from ( a , where it is known. The model predicts that attachment will be significantly different between the two cell types, but viability and cell size will remain unchanged. c Effects of application of channel inhibitors to HOM MEFs in vitro showing application of 10 µM DIDs for 72 h, resulting in decreased cellular attachment (left), and application of 10 nM EIPA ± 10 µM DIDs (right). Data are technical replicates. Dotted lines represent calculated Bliss independence values. d Effects of application of 10 µM DIDs ± 10 nM EIPA on viability in HOM MEFs. e Effects of application of 10 µM DIDs ± 10 nM AHCL on attachment in HOM MEFs. f Effects of application of 10 µM DIDs ± 10 nM AHCL on viability in HOM MEFs.* P
Figure Legend Snippet: Application of the qualitative network to murine embryonic fibroblasts. Expansion of the QN to p53 −/− , Kras G12D/+ (HET) and p53 −/− , Kras G12D/G12D (HOM) MEFs. a Transport proteins deregulated within HOM MEFs when compared to HET MEFs. Proteins are upregulated (red arrows), downregulated (blue arrows), or unchanged (grey). b Phenotype change for MEFS when HOM MEFs are compared to HET MEFs. The model output represents the physiological behaviour predicted by the model when proteins from ( a , where it is known. The model predicts that attachment will be significantly different between the two cell types, but viability and cell size will remain unchanged. c Effects of application of channel inhibitors to HOM MEFs in vitro showing application of 10 µM DIDs for 72 h, resulting in decreased cellular attachment (left), and application of 10 nM EIPA ± 10 µM DIDs (right). Data are technical replicates. Dotted lines represent calculated Bliss independence values. d Effects of application of 10 µM DIDs ± 10 nM EIPA on viability in HOM MEFs. e Effects of application of 10 µM DIDs ± 10 nM AHCL on attachment in HOM MEFs. f Effects of application of 10 µM DIDs ± 10 nM AHCL on viability in HOM MEFs.* P

Techniques Used: In Vitro, Cell Attachment Assay

3) Product Images from "Computational Analysis of Amiloride Analogue Inhibitors of Coxsackievirus B3 RNA Polymerase"

Article Title: Computational Analysis of Amiloride Analogue Inhibitors of Coxsackievirus B3 RNA Polymerase

Journal: Journal of proteomics & bioinformatics

doi: 10.4172/jpb.S9-004

Inhibitor docking to 3D pol All active compounds interact in the same general orientation in the nucleotide binding site. The residues involved in significant interactions with inhibitors are shown (grey sticks) with polar interactions indicated as yellow dashed lines. The schematic 2D representations shown in the right column were created by LigPlot + ] with hydrophobic interactions shown as red semi-circles with lines radiating out in the direction of the interaction. (a) Amiloride (magenta carbons); (b) DMA (orange carbons); (c) EIPA (cyan carbons); (d) HMA (green carbons); (e) Benzamil (yellow carbons); (f) DCB (peach carbons); and (g) HMB (purple carbons).
Figure Legend Snippet: Inhibitor docking to 3D pol All active compounds interact in the same general orientation in the nucleotide binding site. The residues involved in significant interactions with inhibitors are shown (grey sticks) with polar interactions indicated as yellow dashed lines. The schematic 2D representations shown in the right column were created by LigPlot + ] with hydrophobic interactions shown as red semi-circles with lines radiating out in the direction of the interaction. (a) Amiloride (magenta carbons); (b) DMA (orange carbons); (c) EIPA (cyan carbons); (d) HMA (green carbons); (e) Benzamil (yellow carbons); (f) DCB (peach carbons); and (g) HMB (purple carbons).

Techniques Used: Binding Assay

4) Product Images from "Salt secretion is linked to acid-base regulation of ionocytes in seawater-acclimated medaka: new insights into the salt-secreting mechanism"

Article Title: Salt secretion is linked to acid-base regulation of ionocytes in seawater-acclimated medaka: new insights into the salt-secreting mechanism

Journal: Scientific Reports

doi: 10.1038/srep31433

Effects of DIDS (400 μM) ( A ), AZ (200 μM) ( B ), EIPA (400 μM) ( C ), and bumetanide (Bumex; 400 μM) and/or DIDS (400 μM) ( D ) on Cl − flux at ionocytes of larval skin. Data are presented as the means ± SEM. * p
Figure Legend Snippet: Effects of DIDS (400 μM) ( A ), AZ (200 μM) ( B ), EIPA (400 μM) ( C ), and bumetanide (Bumex; 400 μM) and/or DIDS (400 μM) ( D ) on Cl − flux at ionocytes of larval skin. Data are presented as the means ± SEM. * p

Techniques Used:

5) Product Images from "Vaccinia virus strains use distinct forms of macropinocytosis for host-cell entry"

Article Title: Vaccinia virus strains use distinct forms of macropinocytosis for host-cell entry

Journal: Proceedings of the National Academy of Sciences of the United States of America

doi: 10.1073/pnas.1004618107

IHD-J MVs enter cells by PS-mediated macropinocytosis. ( A–C ) Cells were pretreated with staurosporine (Stau), genistein (Geni), wortmannin (Wort), rottlerin (Rott), Gö6979, IPA-3 and PIR 3.5, or EIPA and infected with IHD-J- or WR-EGFP-MVs.
Figure Legend Snippet: IHD-J MVs enter cells by PS-mediated macropinocytosis. ( A–C ) Cells were pretreated with staurosporine (Stau), genistein (Geni), wortmannin (Wort), rottlerin (Rott), Gö6979, IPA-3 and PIR 3.5, or EIPA and infected with IHD-J- or WR-EGFP-MVs.

Techniques Used: Indirect Immunoperoxidase Assay, Infection

6) Product Images from "Hyperosmotic Stress Induces Nuclear Factor-?B Activation and Interleukin-8 Production in Human Intestinal Epithelial Cells"

Article Title: Hyperosmotic Stress Induces Nuclear Factor-?B Activation and Interleukin-8 Production in Human Intestinal Epithelial Cells

Journal: The American Journal of Pathology

doi:

Treatment of HT-29 cells with the selective NHE inhibitors amiloride ( A ), MIA ( B ), or EIPA ( C ) suppresses mannitol-induced IL-8 production. Treatment of HT-29 cells with the nonamiloride NHE inhibitors cimetidine ( D ), clonidine ( E ), or harmaline ( F ), reproduces the suppressive effect of selective NHE inhibitors on the production of IL-8 by mannitol-induced HT-29 cells. When harmaline was used, supernatants for IL-8 measurement were taken 4 hours after the hyperosmotic challenge. In the case of amiloride, MIA, EIPA, cimetidine, and clonidine, IL-8 levels were measured from supernatants obtained 18 hours after the hyperosmotic challenge. Hyperosmotic medium prepared by the addition of 100 mmol/L of mannitol to isosmolar growth medium was used for hyperosmotic stimulation. Data are mean ± SEM of n = 6 to 12 wells from two different experiments. *, P
Figure Legend Snippet: Treatment of HT-29 cells with the selective NHE inhibitors amiloride ( A ), MIA ( B ), or EIPA ( C ) suppresses mannitol-induced IL-8 production. Treatment of HT-29 cells with the nonamiloride NHE inhibitors cimetidine ( D ), clonidine ( E ), or harmaline ( F ), reproduces the suppressive effect of selective NHE inhibitors on the production of IL-8 by mannitol-induced HT-29 cells. When harmaline was used, supernatants for IL-8 measurement were taken 4 hours after the hyperosmotic challenge. In the case of amiloride, MIA, EIPA, cimetidine, and clonidine, IL-8 levels were measured from supernatants obtained 18 hours after the hyperosmotic challenge. Hyperosmotic medium prepared by the addition of 100 mmol/L of mannitol to isosmolar growth medium was used for hyperosmotic stimulation. Data are mean ± SEM of n = 6 to 12 wells from two different experiments. *, P

Techniques Used:

Related Articles

Synthesized:

Article Title: Computational Analysis of Amiloride Analogue Inhibitors of Coxsackievirus B3 RNA Polymerase
Article Snippet: Amiloride, EIPA, 5-( N , N -dimethyl)amiloride (DMA), 5-( N , N -hexamethylene)amiloride (HMA), benzamil and 3’,4’-dichlorobenzamil (DCB) were purchased from Sigma. .. N -Methyl-benzamil (NMB) and methyl 3-amino-5-hexamethyleneimino-6-chloropyrazinoate were synthesized as described in [ ].

Incubation:

Article Title: Orthobunyavirus Entry into Neurons and Other Mammalian Cells Occurs via Clathrin-Mediated Endocytosis and Requires Trafficking into Early Endosomes
Article Snippet: Stock solutions were prepared as follows: 20 mM dynasore (DYN; Tocris Bioscience, MO), 5 mg/ml nystatin (EMD Chemicals, NJ), 50 mM 5-ethylisopropyl amiloride (EIPA; Sigma), 1 mM latrunculin A (EMD Millipore), 1 mM jasplakinolide (EMD Millipore), and 1 mg/ml cytochalasin D (EMD Millipore) were prepared in dimethyl sulfoxide (DMSO), while 0.01 g/ml chlorpromazine (CPZ; Sigma) was prepared in water. .. Cell death did not exceed 5% after incubation of cell cultures with different inhibitors at the indicated working concentrations, as determined by the trypan blue exclusion method.

Article Title: Exploring the role of stromal osmoregulation in cancer and disease using executable modelling
Article Snippet: .. Cells were then treated with indicated concentrations of DIDS, EIPA, AHCL or a combination of these and cell viability was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT, Sigma) assay after 72 h. Cells were incubated with 0.5 mg/ml MTT solution for 3 h at 37 °C before stain was resuspended in DMSO and absorbance at 570 nm determined by microplate reader (Tecan). .. 5000 Murine primary pmFRCs were reversely transfected with 160 ng esiRNA (Sigma) and seeded in quadriplicate into a 96-well plate.

Article Title: The Na+/H+ exchanger contributes to increased smooth muscle proliferation and migration in a rat model of pulmonary arterial hypertension, The Na+/H+ exchanger contributes to increased smooth muscle proliferation and migration in a rat model of pulmonary arterial hypertension
Article Snippet: .. Either 10 μ mol/L EIPA or vehicle (DMSO) was added to the media 1 h after cell seeding, and the cells were incubated for a subsequent 24 h. PASMCs were then fixed in 95% ethanol for 10 min, washed with PBS, and stained with brilliant blue (Sigma) for 5 min. .. Cells were visualized via a microscope‐mounted camera and Q‐capture software.

Article Title: Vitamin D-binding protein controls T cell responses to vitamin D
Article Snippet: For microscopy, cells were incubated with DBP-AF488 for 12 h at 37°C and then stained with anti-CD3 (UCHT1, BD) followed by an AlexaFluor568 coupled anti-mouse Ig and DAPI (nuclear staining). .. Hereafter, the cells were adjusted to 1 × 106 cells/ml, pre-treated with either DMSO or 50 μM EIPA dissolved in DMSO for 30 min and then treated with 30 nM phorbol 12,13-dibutyrate (PDBu) (Sigma-Aldrich) for 60 min as previously described [ , ].

Expressing:

Article Title: Vitamin D-binding protein controls T cell responses to vitamin D
Article Snippet: Hereafter, the cells were adjusted to 1 × 106 cells/ml, pre-treated with either DMSO or 50 μM EIPA dissolved in DMSO for 30 min and then treated with 30 nM phorbol 12,13-dibutyrate (PDBu) (Sigma-Aldrich) for 60 min as previously described [ , ]. .. The TCR surface expression levels were subsequently determined by flow cytometry.

Transfection:

Article Title: Orthobunyavirus Entry into Neurons and Other Mammalian Cells Occurs via Clathrin-Mediated Endocytosis and Requires Trafficking into Early Endosomes
Article Snippet: Stock solutions were prepared as follows: 20 mM dynasore (DYN; Tocris Bioscience, MO), 5 mg/ml nystatin (EMD Chemicals, NJ), 50 mM 5-ethylisopropyl amiloride (EIPA; Sigma), 1 mM latrunculin A (EMD Millipore), 1 mM jasplakinolide (EMD Millipore), and 1 mg/ml cytochalasin D (EMD Millipore) were prepared in dimethyl sulfoxide (DMSO), while 0.01 g/ml chlorpromazine (CPZ; Sigma) was prepared in water. .. Short interfering RNA (siRNA) were from Ambion (Austin, TX), and transfections were performed using Lipofectamine 2000 reagent (Invitrogen, CA) according to the manufacturer's instructions. siRNA samples were used at 33 nM in each transfection (HeLa cells were transfected twice; the second transfection was performed 24 h after the first).

Article Title: Exploring the role of stromal osmoregulation in cancer and disease using executable modelling
Article Snippet: 1000 Murine primary pmFRCs were reversely transfected with 100 ng esiRNA (Sigma) and seeded in triplicates into a 96-well plate. .. Cells were then treated with indicated concentrations of DIDS, EIPA, AHCL or a combination of these and cell viability was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT, Sigma) assay after 72 h. Cells were incubated with 0.5 mg/ml MTT solution for 3 h at 37 °C before stain was resuspended in DMSO and absorbance at 570 nm determined by microplate reader (Tecan).

esiRNA:

Article Title: Exploring the role of stromal osmoregulation in cancer and disease using executable modelling
Article Snippet: 1000 Murine primary pmFRCs were reversely transfected with 100 ng esiRNA (Sigma) and seeded in triplicates into a 96-well plate. .. Cells were then treated with indicated concentrations of DIDS, EIPA, AHCL or a combination of these and cell viability was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT, Sigma) assay after 72 h. Cells were incubated with 0.5 mg/ml MTT solution for 3 h at 37 °C before stain was resuspended in DMSO and absorbance at 570 nm determined by microplate reader (Tecan).

Cell Culture:

Article Title: Cystic fibrosis transmembrane conductance regulator is involved in airway epithelial wound repair
Article Snippet: Calu-3 cell culture media, fetal bovine serum (FBS), penicillin-streptomycin, nonessential amino acids, trypsin, and phosphate-buffered saline (PBS) were purchased from GIBCO (Invitrogen, Carlsbad, CA). .. Trypan blue, EIPA, and cytochalasin D were purchased from Sigma Aldrich (St. Louis, MO).

Indirect Immunoperoxidase Assay:

Article Title: Vaccinia virus strains use distinct forms of macropinocytosis for host-cell entry
Article Snippet: Bafilomycin A1, monensin A, Clostridium difficile toxin B, cytochalasin D, genistein, staurosporine, rottlerin, wortmannin, EIPA, dynasore, chlorpromazine, and cycloheximide were obtained from Sigma-Aldrich, jasplakinolide from Invitrogen, Gö 6979 and 324674 from Calbiochem, Iressa from LC Laboratories, Rhodamine phalloidin from Molecular Probes, and Draq5 from Biostatus Limited. .. IPA-3 and PIR 3.5 were obtained from Jeffery Peterson (Fox Chase Cancer Center, Philadelphia, PA).

Generated:

Article Title: Vaccinia virus strains use distinct forms of macropinocytosis for host-cell entry
Article Snippet: Wild-type (IHD-J and WR), XFP-expressing (IHD-J/WR-EGFP/mRFP-MV), and EGFP-tagged A5 (IHD-J/WR-EGFP-CORE) VACV were generated, purified, and titered as previously described ( ). .. Bafilomycin A1, monensin A, Clostridium difficile toxin B, cytochalasin D, genistein, staurosporine, rottlerin, wortmannin, EIPA, dynasore, chlorpromazine, and cycloheximide were obtained from Sigma-Aldrich, jasplakinolide from Invitrogen, Gö 6979 and 324674 from Calbiochem, Iressa from LC Laboratories, Rhodamine phalloidin from Molecular Probes, and Draq5 from Biostatus Limited.

MTT Assay:

Article Title: Exploring the role of stromal osmoregulation in cancer and disease using executable modelling
Article Snippet: .. Cells were then treated with indicated concentrations of DIDS, EIPA, AHCL or a combination of these and cell viability was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT, Sigma) assay after 72 h. Cells were incubated with 0.5 mg/ml MTT solution for 3 h at 37 °C before stain was resuspended in DMSO and absorbance at 570 nm determined by microplate reader (Tecan). .. 5000 Murine primary pmFRCs were reversely transfected with 160 ng esiRNA (Sigma) and seeded in quadriplicate into a 96-well plate.

Article Title: Hyperosmotic Stress Induces Nuclear Factor-?B Activation and Interleukin-8 Production in Human Intestinal Epithelial Cells
Article Snippet: Amiloride HCl, 5-( N -ethyl- N -isopropyl)-amiloride (EIPA), 5-( N -methyl- N -isobutyl)amiloride (MIA), urea, mannitol, sucrose, and MTT, as well as the nonselective NHE inhibitors cimetidine, harmaline, and clonidine were purchased from Sigma (St. Louis, MO). .. Stock solutions of SB203580, PD98059, amiloride, EIPA, MIA, cimetidine, harmaline, and clonidine were prepared using dimethyl sulfoxide (DMSO, Sigma).

Fluorescence:

Article Title: Exploring the role of stromal osmoregulation in cancer and disease using executable modelling
Article Snippet: After 6 h of incubation under normal growth conditions, fluorescence intensities were measured at 560EX nm/590EM nm with a microplate reader (Tecan). .. Cells were then treated with indicated concentrations of DIDS, EIPA, AHCL or a combination of these and cell viability was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT, Sigma) assay after 72 h. Cells were incubated with 0.5 mg/ml MTT solution for 3 h at 37 °C before stain was resuspended in DMSO and absorbance at 570 nm determined by microplate reader (Tecan).

Microscopy:

Article Title: The Na+/H+ exchanger contributes to increased smooth muscle proliferation and migration in a rat model of pulmonary arterial hypertension, The Na+/H+ exchanger contributes to increased smooth muscle proliferation and migration in a rat model of pulmonary arterial hypertension
Article Snippet: Either 10 μ mol/L EIPA or vehicle (DMSO) was added to the media 1 h after cell seeding, and the cells were incubated for a subsequent 24 h. PASMCs were then fixed in 95% ethanol for 10 min, washed with PBS, and stained with brilliant blue (Sigma) for 5 min. .. Cells were visualized via a microscope‐mounted camera and Q‐capture software.

Article Title: Vitamin D-binding protein controls T cell responses to vitamin D
Article Snippet: For microscopy, cells were incubated with DBP-AF488 for 12 h at 37°C and then stained with anti-CD3 (UCHT1, BD) followed by an AlexaFluor568 coupled anti-mouse Ig and DAPI (nuclear staining). .. Hereafter, the cells were adjusted to 1 × 106 cells/ml, pre-treated with either DMSO or 50 μM EIPA dissolved in DMSO for 30 min and then treated with 30 nM phorbol 12,13-dibutyrate (PDBu) (Sigma-Aldrich) for 60 min as previously described [ , ].

Titration:

Article Title: Extracellular and Luminal pH Regulation by Vacuolar H+-ATPase Isoform Expression and Targeting to the Plasma Membrane and Endosomes *
Article Snippet: Titration curves were linear across the pH range spanned by the experiments. .. Bafilomycin A1 , concanamycin A, and ethylisopropylamiloride (Sigma) were introduced from freshly made stock solutions in DMSO.

Small Interfering RNA:

Article Title: Orthobunyavirus Entry into Neurons and Other Mammalian Cells Occurs via Clathrin-Mediated Endocytosis and Requires Trafficking into Early Endosomes
Article Snippet: Stock solutions were prepared as follows: 20 mM dynasore (DYN; Tocris Bioscience, MO), 5 mg/ml nystatin (EMD Chemicals, NJ), 50 mM 5-ethylisopropyl amiloride (EIPA; Sigma), 1 mM latrunculin A (EMD Millipore), 1 mM jasplakinolide (EMD Millipore), and 1 mg/ml cytochalasin D (EMD Millipore) were prepared in dimethyl sulfoxide (DMSO), while 0.01 g/ml chlorpromazine (CPZ; Sigma) was prepared in water. .. Short interfering RNA (siRNA) were from Ambion (Austin, TX), and transfections were performed using Lipofectamine 2000 reagent (Invitrogen, CA) according to the manufacturer's instructions. siRNA samples were used at 33 nM in each transfection (HeLa cells were transfected twice; the second transfection was performed 24 h after the first).

Confocal Microscopy:

Article Title: Vitamin D-binding protein controls T cell responses to vitamin D
Article Snippet: Hereafter, the cells were adjusted to 1 × 106 cells/ml, pre-treated with either DMSO or 50 μM EIPA dissolved in DMSO for 30 min and then treated with 30 nM phorbol 12,13-dibutyrate (PDBu) (Sigma-Aldrich) for 60 min as previously described [ , ]. .. Hereafter, the cells were adjusted to 1 × 106 cells/ml, pre-treated with either DMSO or 50 μM EIPA dissolved in DMSO for 30 min and then treated with 30 nM phorbol 12,13-dibutyrate (PDBu) (Sigma-Aldrich) for 60 min as previously described [ , ].

IA:

Article Title: Ouabain stimulates Na-K-ATPase through a sodium/hydrogen exchanger-1 (NHE-1)-dependent mechanism in human kidney proximal tubule cells
Article Snippet: Ouabain, 8-(diethyl amino) octyo-3,4,5-trimethoxybenzoate (TMB-8), EIPA, and ammonium chloride were purchased from Sigma (St. Louis, MO). .. The monoclonal antibody against the Na-K-ATPase α1-subunit (α6F) developed by Dr. D. M. Fambrough was obtained from the Developmental Studies Hybridoma Bank, developed under the auspices of NIH CD and maintained by the University of Iowa, Department of Biological Sciences (Iowa City, IA).

Purification:

Article Title: Vaccinia virus strains use distinct forms of macropinocytosis for host-cell entry
Article Snippet: Wild-type (IHD-J and WR), XFP-expressing (IHD-J/WR-EGFP/mRFP-MV), and EGFP-tagged A5 (IHD-J/WR-EGFP-CORE) VACV were generated, purified, and titered as previously described ( ). .. Bafilomycin A1, monensin A, Clostridium difficile toxin B, cytochalasin D, genistein, staurosporine, rottlerin, wortmannin, EIPA, dynasore, chlorpromazine, and cycloheximide were obtained from Sigma-Aldrich, jasplakinolide from Invitrogen, Gö 6979 and 324674 from Calbiochem, Iressa from LC Laboratories, Rhodamine phalloidin from Molecular Probes, and Draq5 from Biostatus Limited.

Viability Assay:

Article Title: Exploring the role of stromal osmoregulation in cancer and disease using executable modelling
Article Snippet: Twenty-four hours later the medium was exchanged and 72 h later cell viability was measured by the addition of CellTiter-Blue (Promega) viability assay reagent. .. Cells were then treated with indicated concentrations of DIDS, EIPA, AHCL or a combination of these and cell viability was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT, Sigma) assay after 72 h. Cells were incubated with 0.5 mg/ml MTT solution for 3 h at 37 °C before stain was resuspended in DMSO and absorbance at 570 nm determined by microplate reader (Tecan).

Plasmid Preparation:

Article Title: Enhancing Endosomal Escape for Intracellular Delivery of Macromolecular Biologic Therapeutics
Article Snippet: Plasmids, antibodies, siRNAs and other reagents Mammalian optimized pCMV-mGFPβ1-10 plasmid (22004005) was purchased from Sandia Biotech. .. EIPA, Chloroquine, and DMSO were from Sigma.

Software:

Article Title: The Na+/H+ exchanger contributes to increased smooth muscle proliferation and migration in a rat model of pulmonary arterial hypertension, The Na+/H+ exchanger contributes to increased smooth muscle proliferation and migration in a rat model of pulmonary arterial hypertension
Article Snippet: Either 10 μ mol/L EIPA or vehicle (DMSO) was added to the media 1 h after cell seeding, and the cells were incubated for a subsequent 24 h. PASMCs were then fixed in 95% ethanol for 10 min, washed with PBS, and stained with brilliant blue (Sigma) for 5 min. .. Cells were visualized via a microscope‐mounted camera and Q‐capture software.

Concentration Assay:

Article Title: Extracellular and Luminal pH Regulation by Vacuolar H+-ATPase Isoform Expression and Targeting to the Plasma Membrane and Endosomes *
Article Snippet: Rates of proton efflux (J H + , in nm H+ min−1 ) were calculated by first converting ECAR into a rate of change of pH (using the Nernst equation) and then calculating the increase in H+ concentration assuming a starting concentration of 40 nm , pH 7.4. .. Bafilomycin A1 , concanamycin A, and ethylisopropylamiloride (Sigma) were introduced from freshly made stock solutions in DMSO.

Article Title: Salt secretion is linked to acid-base regulation of ionocytes in seawater-acclimated medaka: new insights into the salt-secreting mechanism
Article Snippet: Addition of DIDS, AZ, EIPA, and Bumetanide 4, 4′-diisothiocyanatostilbene-2, 2′-disulphonic acid (DIDS; 400 μM), acetazolamide (AZ; 200 μM), 5-(N-ethyl-N-isopropyl)-amiloride (EIPA; 400 μM), and bumetanide (Bumex; 400 μM) were obtained from Sigma-Aldrich, and were used to respectively suppress AE, CA, NHE, and NKCC in this study. .. The final concentration of DMSO or ethanol in the working solutions (including the control group) was 0.1%.

Cell Counting:

Article Title: The Na+/H+ exchanger contributes to increased smooth muscle proliferation and migration in a rat model of pulmonary arterial hypertension, The Na+/H+ exchanger contributes to increased smooth muscle proliferation and migration in a rat model of pulmonary arterial hypertension
Article Snippet: Either 10 μ mol/L EIPA or vehicle (DMSO) was added to the media 1 h after cell seeding, and the cells were incubated for a subsequent 24 h. PASMCs were then fixed in 95% ethanol for 10 min, washed with PBS, and stained with brilliant blue (Sigma) for 5 min. .. For each filter, five randomly chosen fields were imaged to obtain a total adherent cell count.

Migration:

Article Title: The Na+/H+ exchanger contributes to increased smooth muscle proliferation and migration in a rat model of pulmonary arterial hypertension, The Na+/H+ exchanger contributes to increased smooth muscle proliferation and migration in a rat model of pulmonary arterial hypertension
Article Snippet: Paragraph title: PASMC migration ... Either 10 μ mol/L EIPA or vehicle (DMSO) was added to the media 1 h after cell seeding, and the cells were incubated for a subsequent 24 h. PASMCs were then fixed in 95% ethanol for 10 min, washed with PBS, and stained with brilliant blue (Sigma) for 5 min.

Staining:

Article Title: SOD1 protein aggregates stimulate macropinocytosis in neurons to facilitate their propagation
Article Snippet: EIPA, CPZ, Gen, and Rot were purchased from Sigma Aldrich. .. Alexa Fluor 488 goat anti-mouse, Alexa Fluor 488 goat anti-rabbit, streptavidin Alexa Fluor 633 conjugate, streptavidin Alexa Fluor 488 conjugate, Alexa Fluor 488 donkey anti-sheep, Alexa Fluor 488 donkey anti-rabbit, SYTOX Red dead cell stain, FM® 1-43FX fixable analogue of FM® 1–43 membrane stain were purchased from Invitrogen Life Technologies.

Article Title: Exploring the role of stromal osmoregulation in cancer and disease using executable modelling
Article Snippet: .. Cells were then treated with indicated concentrations of DIDS, EIPA, AHCL or a combination of these and cell viability was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT, Sigma) assay after 72 h. Cells were incubated with 0.5 mg/ml MTT solution for 3 h at 37 °C before stain was resuspended in DMSO and absorbance at 570 nm determined by microplate reader (Tecan). .. 5000 Murine primary pmFRCs were reversely transfected with 160 ng esiRNA (Sigma) and seeded in quadriplicate into a 96-well plate.

Article Title: The Na+/H+ exchanger contributes to increased smooth muscle proliferation and migration in a rat model of pulmonary arterial hypertension, The Na+/H+ exchanger contributes to increased smooth muscle proliferation and migration in a rat model of pulmonary arterial hypertension
Article Snippet: .. Either 10 μ mol/L EIPA or vehicle (DMSO) was added to the media 1 h after cell seeding, and the cells were incubated for a subsequent 24 h. PASMCs were then fixed in 95% ethanol for 10 min, washed with PBS, and stained with brilliant blue (Sigma) for 5 min. .. Cells were visualized via a microscope‐mounted camera and Q‐capture software.

Article Title: Vitamin D-binding protein controls T cell responses to vitamin D
Article Snippet: For microscopy, cells were incubated with DBP-AF488 for 12 h at 37°C and then stained with anti-CD3 (UCHT1, BD) followed by an AlexaFluor568 coupled anti-mouse Ig and DAPI (nuclear staining). .. Hereafter, the cells were adjusted to 1 × 106 cells/ml, pre-treated with either DMSO or 50 μM EIPA dissolved in DMSO for 30 min and then treated with 30 nM phorbol 12,13-dibutyrate (PDBu) (Sigma-Aldrich) for 60 min as previously described [ , ].

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    Millipore macropinocytosis inhibitor eipa
    Analyses of the role of endocytosis routes and paracellular permeability in antibody transfer to the draining LNs. ( A ) Confocal and flow cytometric analyses of draining LNs of WT and Cav1 −/− mice after s.c. administration of the indicated fluorochrome-conjugated antibodies (1 μg, t = 5 min, n = 4). The cells were stained ex vivo for CD3 for flow cytometric analysis. Scale bars: 20 μm. ( B ) Experimental outline for topical application of endocytosis inhibitors (and controls) to surgically exposed LNs. ( C – G ) Analyses of transcytosis of the indicated s.c. administered fluorochrome-conjugated antibodies (5 μg each, t = 5 min) after topical pretreatment of the draining LNs with ( C – F ) endocytosis inhibitors or ( G ) a LEC junctional stabilizer. ( C ) Pitstop 2 (a clathrin inhibitor) and a Pitstop 2 negative control; ( D ) monodansylcadaverine (MDC) (a clathrin inhibitor) and vehicle; ( E ) <t>EIPA</t> (a <t>macropinocytosis</t> inhibitor) and vehicle; ( F ) imipramine (a macropinocytosis inhibitor) and vehicle; and ( G ) adrenomedullin and vehicle. The cells were stained ex vivo for CD3. In the bar graphs, each dot represents 1 LN, and data are the mean ± SD. Statistical significance was determined by Mann-Whitney U test.
    Macropinocytosis Inhibitor Eipa, supplied by Millipore, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    77
    Millipore sodium proton exchanger inhibitor eipa
    Inhibitors against Pak1, <t>PKC,</t> sodium/proton exchanger, and actin reduce Ad35-eGFP transduction. HeLa-ATCC and HK-2 cells were preincubated with the indicated concentrations of drugs for 30 min and infected with Ad35-eGFP for 8 h. Cells were analyzed on a Safire2 plate reader, and eGFP intensities were normalized to the DAPI signal of the cell nuclei, representing cell numbers. Results for <t>EIPA</t> are shown in panel A, NH 4 Cl in B, bafilomycin (Baf) in C, jasplakinolide (Jas) and cytochalasin D (CytD) in D, the PKC inhibitor Gö6976 in E, and the Pak1 inhibitor IPA-3 and its inactive derivative PIR3.5 in F and G for both Ad35-eGFP and Ad5-eGFP, respectively. Panels H to K show results with macropinocytic interference in Wi-38 cells transduced with Ad35-eGFP or Ad5-eGFP. Note the absence of significant cell toxicity, as indicated by cell number measurements shown with gray line graphs, with 100% representing noninfected non-drug-treated conditions.
    Sodium Proton Exchanger Inhibitor Eipa, supplied by Millipore, used in various techniques. Bioz Stars score: 77/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Price from $9.99 to $1999.99
    sodium proton exchanger inhibitor eipa - by Bioz Stars, 2020-02
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    Analyses of the role of endocytosis routes and paracellular permeability in antibody transfer to the draining LNs. ( A ) Confocal and flow cytometric analyses of draining LNs of WT and Cav1 −/− mice after s.c. administration of the indicated fluorochrome-conjugated antibodies (1 μg, t = 5 min, n = 4). The cells were stained ex vivo for CD3 for flow cytometric analysis. Scale bars: 20 μm. ( B ) Experimental outline for topical application of endocytosis inhibitors (and controls) to surgically exposed LNs. ( C – G ) Analyses of transcytosis of the indicated s.c. administered fluorochrome-conjugated antibodies (5 μg each, t = 5 min) after topical pretreatment of the draining LNs with ( C – F ) endocytosis inhibitors or ( G ) a LEC junctional stabilizer. ( C ) Pitstop 2 (a clathrin inhibitor) and a Pitstop 2 negative control; ( D ) monodansylcadaverine (MDC) (a clathrin inhibitor) and vehicle; ( E ) EIPA (a macropinocytosis inhibitor) and vehicle; ( F ) imipramine (a macropinocytosis inhibitor) and vehicle; and ( G ) adrenomedullin and vehicle. The cells were stained ex vivo for CD3. In the bar graphs, each dot represents 1 LN, and data are the mean ± SD. Statistical significance was determined by Mann-Whitney U test.

    Journal: The Journal of Clinical Investigation

    Article Title: Transcytosis route mediates rapid delivery of intact antibodies to draining lymph nodes

    doi: 10.1172/JCI125740

    Figure Lengend Snippet: Analyses of the role of endocytosis routes and paracellular permeability in antibody transfer to the draining LNs. ( A ) Confocal and flow cytometric analyses of draining LNs of WT and Cav1 −/− mice after s.c. administration of the indicated fluorochrome-conjugated antibodies (1 μg, t = 5 min, n = 4). The cells were stained ex vivo for CD3 for flow cytometric analysis. Scale bars: 20 μm. ( B ) Experimental outline for topical application of endocytosis inhibitors (and controls) to surgically exposed LNs. ( C – G ) Analyses of transcytosis of the indicated s.c. administered fluorochrome-conjugated antibodies (5 μg each, t = 5 min) after topical pretreatment of the draining LNs with ( C – F ) endocytosis inhibitors or ( G ) a LEC junctional stabilizer. ( C ) Pitstop 2 (a clathrin inhibitor) and a Pitstop 2 negative control; ( D ) monodansylcadaverine (MDC) (a clathrin inhibitor) and vehicle; ( E ) EIPA (a macropinocytosis inhibitor) and vehicle; ( F ) imipramine (a macropinocytosis inhibitor) and vehicle; and ( G ) adrenomedullin and vehicle. The cells were stained ex vivo for CD3. In the bar graphs, each dot represents 1 LN, and data are the mean ± SD. Statistical significance was determined by Mann-Whitney U test.

    Article Snippet: The following endocytosis inhibitors and controls were used: clathrin inhibitor Pitstop 2 ( ) (Abcam) and its specific negative control (Pitstop 2 control, Abcam) both at 75 μM; the clathrin inhibitor monodansylcadaverine ( ) (MilliporeSigma; 100 μM) and its DMSO vehicle control; the macropinocytosis inhibitor EIPA ( ) [5-( N -ethyl- N -isopropyl amiloride, MilliporeSigma; 100 μM] and its DMSO vehicle control; the macropinocytosis inhibitor imipramine ( ) (MilliporeSigma, 15 μM) and its PBS vehicle control; the dynamin inhibitor Dyngo-4a ( ) (MilliporeSigma, 400 μM) and its DMSO vehicle control; and the dynamin inhibitor Dynole 34-2 (39 (Abcam) and its specific control Dynole 31-2 (Abcam), both at 75 μM.

    Techniques: Permeability, Flow Cytometry, Mouse Assay, Staining, Ex Vivo, Negative Control, MANN-WHITNEY

    Inhibitors against Pak1, PKC, sodium/proton exchanger, and actin reduce Ad35-eGFP transduction. HeLa-ATCC and HK-2 cells were preincubated with the indicated concentrations of drugs for 30 min and infected with Ad35-eGFP for 8 h. Cells were analyzed on a Safire2 plate reader, and eGFP intensities were normalized to the DAPI signal of the cell nuclei, representing cell numbers. Results for EIPA are shown in panel A, NH 4 Cl in B, bafilomycin (Baf) in C, jasplakinolide (Jas) and cytochalasin D (CytD) in D, the PKC inhibitor Gö6976 in E, and the Pak1 inhibitor IPA-3 and its inactive derivative PIR3.5 in F and G for both Ad35-eGFP and Ad5-eGFP, respectively. Panels H to K show results with macropinocytic interference in Wi-38 cells transduced with Ad35-eGFP or Ad5-eGFP. Note the absence of significant cell toxicity, as indicated by cell number measurements shown with gray line graphs, with 100% representing noninfected non-drug-treated conditions.

    Journal: Journal of Virology

    Article Title: Macropinocytotic Uptake and Infection of Human Epithelial Cells with Species B2 Adenovirus Type 35 ▿Macropinocytotic Uptake and Infection of Human Epithelial Cells with Species B2 Adenovirus Type 35 ▿ †

    doi: 10.1128/JVI.02494-09

    Figure Lengend Snippet: Inhibitors against Pak1, PKC, sodium/proton exchanger, and actin reduce Ad35-eGFP transduction. HeLa-ATCC and HK-2 cells were preincubated with the indicated concentrations of drugs for 30 min and infected with Ad35-eGFP for 8 h. Cells were analyzed on a Safire2 plate reader, and eGFP intensities were normalized to the DAPI signal of the cell nuclei, representing cell numbers. Results for EIPA are shown in panel A, NH 4 Cl in B, bafilomycin (Baf) in C, jasplakinolide (Jas) and cytochalasin D (CytD) in D, the PKC inhibitor Gö6976 in E, and the Pak1 inhibitor IPA-3 and its inactive derivative PIR3.5 in F and G for both Ad35-eGFP and Ad5-eGFP, respectively. Panels H to K show results with macropinocytic interference in Wi-38 cells transduced with Ad35-eGFP or Ad5-eGFP. Note the absence of significant cell toxicity, as indicated by cell number measurements shown with gray line graphs, with 100% representing noninfected non-drug-treated conditions.

    Article Snippet: The PKC inhibitor Gö6976, the sodium/proton exchanger inhibitor EIPA, cytochalasin D, and jasplakinolide were from Calbiochem.

    Techniques: Transduction, Infection, Indirect Immunoperoxidase Assay