Structured Review

Abcam k atpase
Profiling of membrane-fraction proteins from the prefrontal cortex of control and alcoholic subjects. Control (C) or alcoholic (A) prefrontal cortex membrane fraction proteins were resolved by 1D PAGE and proteins stained with colloidal Coomassie. Alcoholic subjects displayed a prominent reduction in the level of protein staining at a protein band of ∼270 <t>kDa</t> and upregulation at ∼112 kDa (upper panel). Protein levels of transmembranous (∼112 kDa) Na + ,K + <t>-ATPase</t> α-subunit, and oligomeric forms of the α-subunit were visualized by Western blotting (middle panel). The level of transmembranous α-subunit was quantified using actin for normalisation (lower panel). For marked significance: *** = p
K Atpase, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Images

1) Product Images from "Alcohol-Related Brain Damage in Humans"

Article Title: Alcohol-Related Brain Damage in Humans

Journal: PLoS ONE

doi: 10.1371/journal.pone.0093586

Profiling of membrane-fraction proteins from the prefrontal cortex of control and alcoholic subjects. Control (C) or alcoholic (A) prefrontal cortex membrane fraction proteins were resolved by 1D PAGE and proteins stained with colloidal Coomassie. Alcoholic subjects displayed a prominent reduction in the level of protein staining at a protein band of ∼270 kDa and upregulation at ∼112 kDa (upper panel). Protein levels of transmembranous (∼112 kDa) Na + ,K + -ATPase α-subunit, and oligomeric forms of the α-subunit were visualized by Western blotting (middle panel). The level of transmembranous α-subunit was quantified using actin for normalisation (lower panel). For marked significance: *** = p
Figure Legend Snippet: Profiling of membrane-fraction proteins from the prefrontal cortex of control and alcoholic subjects. Control (C) or alcoholic (A) prefrontal cortex membrane fraction proteins were resolved by 1D PAGE and proteins stained with colloidal Coomassie. Alcoholic subjects displayed a prominent reduction in the level of protein staining at a protein band of ∼270 kDa and upregulation at ∼112 kDa (upper panel). Protein levels of transmembranous (∼112 kDa) Na + ,K + -ATPase α-subunit, and oligomeric forms of the α-subunit were visualized by Western blotting (middle panel). The level of transmembranous α-subunit was quantified using actin for normalisation (lower panel). For marked significance: *** = p

Techniques Used: Polyacrylamide Gel Electrophoresis, Staining, Western Blot

2) Product Images from "Alcohol-Related Brain Damage in Humans"

Article Title: Alcohol-Related Brain Damage in Humans

Journal: PLoS ONE

doi: 10.1371/journal.pone.0093586

Profiling of membrane-fraction proteins from the prefrontal cortex of control and alcoholic subjects. Control (C) or alcoholic (A) prefrontal cortex membrane fraction proteins were resolved by 1D PAGE and proteins stained with colloidal Coomassie. Alcoholic subjects displayed a prominent reduction in the level of protein staining at a protein band of ∼270 kDa and upregulation at ∼112 kDa (upper panel). Protein levels of transmembranous (∼112 kDa) Na + ,K + -ATPase α-subunit, and oligomeric forms of the α-subunit were visualized by Western blotting (middle panel). The level of transmembranous α-subunit was quantified using actin for normalisation (lower panel). For marked significance: *** = p
Figure Legend Snippet: Profiling of membrane-fraction proteins from the prefrontal cortex of control and alcoholic subjects. Control (C) or alcoholic (A) prefrontal cortex membrane fraction proteins were resolved by 1D PAGE and proteins stained with colloidal Coomassie. Alcoholic subjects displayed a prominent reduction in the level of protein staining at a protein band of ∼270 kDa and upregulation at ∼112 kDa (upper panel). Protein levels of transmembranous (∼112 kDa) Na + ,K + -ATPase α-subunit, and oligomeric forms of the α-subunit were visualized by Western blotting (middle panel). The level of transmembranous α-subunit was quantified using actin for normalisation (lower panel). For marked significance: *** = p

Techniques Used: Polyacrylamide Gel Electrophoresis, Staining, Western Blot

3) Product Images from "Reduced E-cadherin facilitates renal cell carcinoma progression by WNT/β-catenin signaling activation"

Article Title: Reduced E-cadherin facilitates renal cell carcinoma progression by WNT/β-catenin signaling activation

Journal: Oncotarget

doi: 10.18632/oncotarget.15361

Correlation between E-cadherin and WNT/β-catenin signaling in RCC tissues ( A ) E-cadherin, β-catenin and cyclin D1 protein levels were analyzed with western blot in four fresh RCC tissues. β-actin was used as loading control. ( B ) mRNA levels of E-cadherin and WNT/β-catenin signaling targeted genes was studied with RT-PCR in the four fresh RCC tissues. β-actin was used as control. ( C , D ) β-catenin location in E-cadherin positive (C) or negative (D) specimens were studied with immunobloting assays. Na, K ATPase was used as membrane control, and β-actin as cytoplasm control. Representative images from at least three independent experiments are shown.
Figure Legend Snippet: Correlation between E-cadherin and WNT/β-catenin signaling in RCC tissues ( A ) E-cadherin, β-catenin and cyclin D1 protein levels were analyzed with western blot in four fresh RCC tissues. β-actin was used as loading control. ( B ) mRNA levels of E-cadherin and WNT/β-catenin signaling targeted genes was studied with RT-PCR in the four fresh RCC tissues. β-actin was used as control. ( C , D ) β-catenin location in E-cadherin positive (C) or negative (D) specimens were studied with immunobloting assays. Na, K ATPase was used as membrane control, and β-actin as cytoplasm control. Representative images from at least three independent experiments are shown.

Techniques Used: Western Blot, Reverse Transcription Polymerase Chain Reaction

4) Product Images from "Alcohol-Related Brain Damage in Humans"

Article Title: Alcohol-Related Brain Damage in Humans

Journal: PLoS ONE

doi: 10.1371/journal.pone.0093586

Profiling of membrane-fraction proteins from the prefrontal cortex of control and alcoholic subjects. Control (C) or alcoholic (A) prefrontal cortex membrane fraction proteins were resolved by 1D PAGE and proteins stained with colloidal Coomassie. Alcoholic subjects displayed a prominent reduction in the level of protein staining at a protein band of ∼270 kDa and upregulation at ∼112 kDa (upper panel). Protein levels of transmembranous (∼112 kDa) Na + ,K + -ATPase α-subunit, and oligomeric forms of the α-subunit were visualized by Western blotting (middle panel). The level of transmembranous α-subunit was quantified using actin for normalisation (lower panel). For marked significance: *** = p
Figure Legend Snippet: Profiling of membrane-fraction proteins from the prefrontal cortex of control and alcoholic subjects. Control (C) or alcoholic (A) prefrontal cortex membrane fraction proteins were resolved by 1D PAGE and proteins stained with colloidal Coomassie. Alcoholic subjects displayed a prominent reduction in the level of protein staining at a protein band of ∼270 kDa and upregulation at ∼112 kDa (upper panel). Protein levels of transmembranous (∼112 kDa) Na + ,K + -ATPase α-subunit, and oligomeric forms of the α-subunit were visualized by Western blotting (middle panel). The level of transmembranous α-subunit was quantified using actin for normalisation (lower panel). For marked significance: *** = p

Techniques Used: Polyacrylamide Gel Electrophoresis, Staining, Western Blot

5) Product Images from "Alcohol-Related Brain Damage in Humans"

Article Title: Alcohol-Related Brain Damage in Humans

Journal: PLoS ONE

doi: 10.1371/journal.pone.0093586

Profiling of membrane-fraction proteins from the prefrontal cortex of control and alcoholic subjects. Control (C) or alcoholic (A) prefrontal cortex membrane fraction proteins were resolved by 1D PAGE and proteins stained with colloidal Coomassie. Alcoholic subjects displayed a prominent reduction in the level of protein staining at a protein band of ∼270 kDa and upregulation at ∼112 kDa (upper panel). Protein levels of transmembranous (∼112 kDa) Na + ,K + -ATPase α-subunit, and oligomeric forms of the α-subunit were visualized by Western blotting (middle panel). The level of transmembranous α-subunit was quantified using actin for normalisation (lower panel). For marked significance: *** = p
Figure Legend Snippet: Profiling of membrane-fraction proteins from the prefrontal cortex of control and alcoholic subjects. Control (C) or alcoholic (A) prefrontal cortex membrane fraction proteins were resolved by 1D PAGE and proteins stained with colloidal Coomassie. Alcoholic subjects displayed a prominent reduction in the level of protein staining at a protein band of ∼270 kDa and upregulation at ∼112 kDa (upper panel). Protein levels of transmembranous (∼112 kDa) Na + ,K + -ATPase α-subunit, and oligomeric forms of the α-subunit were visualized by Western blotting (middle panel). The level of transmembranous α-subunit was quantified using actin for normalisation (lower panel). For marked significance: *** = p

Techniques Used: Polyacrylamide Gel Electrophoresis, Staining, Western Blot

6) Product Images from "Alcohol-Related Brain Damage in Humans"

Article Title: Alcohol-Related Brain Damage in Humans

Journal: PLoS ONE

doi: 10.1371/journal.pone.0093586

Profiling of membrane-fraction proteins from the prefrontal cortex of control and alcoholic subjects. Control (C) or alcoholic (A) prefrontal cortex membrane fraction proteins were resolved by 1D PAGE and proteins stained with colloidal Coomassie. Alcoholic subjects displayed a prominent reduction in the level of protein staining at a protein band of ∼270 kDa and upregulation at ∼112 kDa (upper panel). Protein levels of transmembranous (∼112 kDa) Na + ,K + -ATPase α-subunit, and oligomeric forms of the α-subunit were visualized by Western blotting (middle panel). The level of transmembranous α-subunit was quantified using actin for normalisation (lower panel). For marked significance: *** = p
Figure Legend Snippet: Profiling of membrane-fraction proteins from the prefrontal cortex of control and alcoholic subjects. Control (C) or alcoholic (A) prefrontal cortex membrane fraction proteins were resolved by 1D PAGE and proteins stained with colloidal Coomassie. Alcoholic subjects displayed a prominent reduction in the level of protein staining at a protein band of ∼270 kDa and upregulation at ∼112 kDa (upper panel). Protein levels of transmembranous (∼112 kDa) Na + ,K + -ATPase α-subunit, and oligomeric forms of the α-subunit were visualized by Western blotting (middle panel). The level of transmembranous α-subunit was quantified using actin for normalisation (lower panel). For marked significance: *** = p

Techniques Used: Polyacrylamide Gel Electrophoresis, Staining, Western Blot

Related Articles

Western Blot:

Article Title: Oxidative Stress Mediates the Disruption of Airway Epithelial Tight Junctions through a TRPM2-PLC?1-PKC? Signaling Pathway
Article Snippet: .. All of the antibodies used for Western blotting, including PLCγ1 antibody (Abcam, ab16955) (Cambridge, MA, USA), phosphorylated PLCγ1 Y783 antibody (Abcam, ab53125), PKCα antibody (Abcam, ab32376), ZO-1 antibody (Abcam, ab59720), claudin-2 antibody (Abcam, ab53032), β-actin antibody (Abcam, ab25894), Na+ -K+ ATPase antibody (Abcam, ab76509), second antibody anti-mouse IgG (Abcam, ab6789) and anti-rabbit IgG (Abcam, ab97200) were purchased from Abcam (Cambridge, MA, USA). .. The transfection reagent FuGENE HD was acquired from Roche (Basel, Switzerland).

Article Title: IL-33 promotes IL-10 production in macrophages: a role for IL-33 in macrophage foam cell formation
Article Snippet: .. For the detection of ST2L presence in the MFC membrane, total membrane proteins were extracted using a CelLytic MEM protein extraction kit (Sigma), and were subsequently subjected to routine western blot analysis, as described above, and anti-ATPase Na+ /K+ -β2, instead of GAPDH, antibody (Abcam) was used as a loading control. .. IL-10 mRNA stability detection and dual-luciferase reporter gene assays IL-10 mRNA is unstable; therefore, stabilization of its mRNA is important for its expression.

Protein Extraction:

Article Title: IL-33 promotes IL-10 production in macrophages: a role for IL-33 in macrophage foam cell formation
Article Snippet: .. For the detection of ST2L presence in the MFC membrane, total membrane proteins were extracted using a CelLytic MEM protein extraction kit (Sigma), and were subsequently subjected to routine western blot analysis, as described above, and anti-ATPase Na+ /K+ -β2, instead of GAPDH, antibody (Abcam) was used as a loading control. .. IL-10 mRNA stability detection and dual-luciferase reporter gene assays IL-10 mRNA is unstable; therefore, stabilization of its mRNA is important for its expression.

Immunocytochemistry:

Article Title: Arresting proliferation improves the cell identity of corneal endothelial cells in the New Zealand rabbit
Article Snippet: .. Immunocytochemistry Immunocytochemistry was performed in CECs before culture, after culture in MitoM, and in RestM to analyze the presence of GPC4 (Abcam, ab150517, Cambridge, UK), CD166 (Abcam, ab78649), ZO-1/TJP1 (ThermoFisher, 61–7300, Waltham, MA), and Na/K-ATPase (Abcam, ab176163, Cambridge, UK). .. Immunocytochemistry consisted of overnight cell stabilization over coverslips with poly-D lysine (Sigma-Aldrich, P7280), fixation with 4% paraformaldehyde, nonspecific bonding blockage with 5% bovine serum albumin (BSA; Sigma-Aldrich, A-7030), overnight 4 °C incubation with primary antibodies (GPC4 5 µg/ml, CD166 1 µg/ml, ZO-1 5 µg/ml, and Na/K-ATPase 1:100), and incubation with Alexa Fluor 488 secondary antibody (Abcam, ab150077) for 1 h at room temperature.

Article Title: Primary explant culture and collagen I substrate enhances corneal endothelial cell morphology
Article Snippet: .. Immunocytochemistry was performed to analyze the presence of GPC4 (Abcam, ab150517, Cambridge, UK) and Na/K-ATPase (Abcam, ab176163, Cambridge, UK) in CEC cultured in each condition. .. Images were obtained with a fluorescence inverted microscope.

Capillary Electrochromatography:

Article Title: Primary explant culture and collagen I substrate enhances corneal endothelial cell morphology
Article Snippet: .. Immunocytochemistry was performed to analyze the presence of GPC4 (Abcam, ab150517, Cambridge, UK) and Na/K-ATPase (Abcam, ab176163, Cambridge, UK) in CEC cultured in each condition. .. Images were obtained with a fluorescence inverted microscope.

Cell Culture:

Article Title: Primary explant culture and collagen I substrate enhances corneal endothelial cell morphology
Article Snippet: .. Immunocytochemistry was performed to analyze the presence of GPC4 (Abcam, ab150517, Cambridge, UK) and Na/K-ATPase (Abcam, ab176163, Cambridge, UK) in CEC cultured in each condition. .. Images were obtained with a fluorescence inverted microscope.

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    Abcam anti α1 na k atpase
    Effect of chemical ischemia on the expression of <t>Na-K</t> <t>ATPase</t> <t>α1</t> subunit and Na-K ATPase activity in cultured astrocytes. (A) Western blot analysis shows the expression of Na-K ATPase α1 subunit protein following chemical ischemia for 1, 2, 3, and 6 h. Data were obtained from five experiments. (B) Real-time PCR analysis shows the expression of Na-K ATPase α1 subunit mRNA in cultured astrocytes following chemical ischemia for 1, 2, 3, and 6 h. Data were obtained from four experiments. (C) The activity of Na-K ATPase in the membrane fraction of cultured astrocytes after chemical ischemia for 1, 2, 3, and 6 h. Data were obtained from five experiments. For all experiments, each value indicates the mean±S.E.M. normalized to the control of each time point. * p
    Anti α1 Na K Atpase, supplied by Abcam, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti α1 na k atpase/product/Abcam
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    99
    Abcam anti na k atpase
    Analysis of wild-type and mutant BSEP distribution by immunofluorescence. The resulting distribution of fluorescence was observed by laser confocal microscopy. Green represents BSEP and red represents calnexin (left) and <t>Na/K-ATPase</t> (right) and yellow represents co-localization of the two proteins. In each panel, an X-Y image (upper) and an X-Z image (lower) are shown. The data shown were obtained from four independent experiments. Scale bar, 5 µm.
    Anti Na K Atpase, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti na k atpase/product/Abcam
    Average 99 stars, based on 10 article reviews
    Price from $9.99 to $1999.99
    anti na k atpase - by Bioz Stars, 2020-05
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    91
    Abcam h k atpase
    (A) Structure of known proton-pump inhibitor, Rabeprazole (B) Gastric <t>H+/K+-ATPase</t> activity is inhibited by 100μM Rabeprazole. (C) Benzimidazole probes inhibit ATPase activity by approximately 50% at 10μM compound.
    H K Atpase, supplied by Abcam, used in various techniques. Bioz Stars score: 91/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/h k atpase/product/Abcam
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    89
    Abcam mouse anti na k atpase
    Localization of ATP-dependent efflux transporter proteins. A-L ) Confocal micrographs after indirect immunofluorescence labeling with efflux pump proteins MRP-1, -4, or -5 (green), and eye-specific proteins MITF and cellular retinaldehyde-binding protein (CRALBP, both red), the polarization marker Na + /K + <t>ATPase</t> (red), and the nuclear label 4′,6′-diamidino-2-phenylidole (blue). In figures M-P ) the brightfield micrographs show the same ARPE-19 cells and fusiform, early cobblestone, and cobblestone hESC-RPE as shown in the confocal images. Scale bars, 10 µm.
    Mouse Anti Na K Atpase, supplied by Abcam, used in various techniques. Bioz Stars score: 89/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Effect of chemical ischemia on the expression of Na-K ATPase α1 subunit and Na-K ATPase activity in cultured astrocytes. (A) Western blot analysis shows the expression of Na-K ATPase α1 subunit protein following chemical ischemia for 1, 2, 3, and 6 h. Data were obtained from five experiments. (B) Real-time PCR analysis shows the expression of Na-K ATPase α1 subunit mRNA in cultured astrocytes following chemical ischemia for 1, 2, 3, and 6 h. Data were obtained from four experiments. (C) The activity of Na-K ATPase in the membrane fraction of cultured astrocytes after chemical ischemia for 1, 2, 3, and 6 h. Data were obtained from five experiments. For all experiments, each value indicates the mean±S.E.M. normalized to the control of each time point. * p

    Journal: The Korean Journal of Physiology & Pharmacology : Official Journal of the Korean Physiological Society and the Korean Society of Pharmacology

    Article Title: Expression and Activity of the Na-K ATPase in Ischemic Injury of Primary Cultured Astrocytes

    doi: 10.4196/kjpp.2013.17.4.275

    Figure Lengend Snippet: Effect of chemical ischemia on the expression of Na-K ATPase α1 subunit and Na-K ATPase activity in cultured astrocytes. (A) Western blot analysis shows the expression of Na-K ATPase α1 subunit protein following chemical ischemia for 1, 2, 3, and 6 h. Data were obtained from five experiments. (B) Real-time PCR analysis shows the expression of Na-K ATPase α1 subunit mRNA in cultured astrocytes following chemical ischemia for 1, 2, 3, and 6 h. Data were obtained from four experiments. (C) The activity of Na-K ATPase in the membrane fraction of cultured astrocytes after chemical ischemia for 1, 2, 3, and 6 h. Data were obtained from five experiments. For all experiments, each value indicates the mean±S.E.M. normalized to the control of each time point. * p

    Article Snippet: The transferred membranes were incubated overnight at 4℃ with primary antibodies including anti-α1 Na-K ATPase (1:5,000, mouse monoclonal; Abcam, San Francisco, CA) and anti-β-actin (1:2,000, mouse monoclonal; Santa Cruz Biotechnology, Paso Robles, CA).

    Techniques: Expressing, Activity Assay, Cell Culture, Western Blot, Real-time Polymerase Chain Reaction

    Primary cultures of astrocytes. Immunohistochemistry of primary astrocytes and Na-K ATPase. DAPI, blue (A); GFAP, red (B); Na-K ATPase, green (C); Merged image (D). Scale bar=200 µm.

    Journal: The Korean Journal of Physiology & Pharmacology : Official Journal of the Korean Physiological Society and the Korean Society of Pharmacology

    Article Title: Expression and Activity of the Na-K ATPase in Ischemic Injury of Primary Cultured Astrocytes

    doi: 10.4196/kjpp.2013.17.4.275

    Figure Lengend Snippet: Primary cultures of astrocytes. Immunohistochemistry of primary astrocytes and Na-K ATPase. DAPI, blue (A); GFAP, red (B); Na-K ATPase, green (C); Merged image (D). Scale bar=200 µm.

    Article Snippet: The transferred membranes were incubated overnight at 4℃ with primary antibodies including anti-α1 Na-K ATPase (1:5,000, mouse monoclonal; Abcam, San Francisco, CA) and anti-β-actin (1:2,000, mouse monoclonal; Santa Cruz Biotechnology, Paso Robles, CA).

    Techniques: Immunohistochemistry

    Effects of reoxygenation on the changes in the cytotoxicity, viability, and activity of the Na-K ATPase. The levels of LDH release (A) and MTT reduction (B) were quantified at 24 h of reoxygenation following chemical ischemia for 1, 2, 3, and 6 h. (C) Activity of Na-K ATPase was measured at 24 h of reoxygenation following chemical ischemia for 1 and 2 h. Data show the mean± S.E.M. of the relative values obtained from five animals. * p

    Journal: The Korean Journal of Physiology & Pharmacology : Official Journal of the Korean Physiological Society and the Korean Society of Pharmacology

    Article Title: Expression and Activity of the Na-K ATPase in Ischemic Injury of Primary Cultured Astrocytes

    doi: 10.4196/kjpp.2013.17.4.275

    Figure Lengend Snippet: Effects of reoxygenation on the changes in the cytotoxicity, viability, and activity of the Na-K ATPase. The levels of LDH release (A) and MTT reduction (B) were quantified at 24 h of reoxygenation following chemical ischemia for 1, 2, 3, and 6 h. (C) Activity of Na-K ATPase was measured at 24 h of reoxygenation following chemical ischemia for 1 and 2 h. Data show the mean± S.E.M. of the relative values obtained from five animals. * p

    Article Snippet: The transferred membranes were incubated overnight at 4℃ with primary antibodies including anti-α1 Na-K ATPase (1:5,000, mouse monoclonal; Abcam, San Francisco, CA) and anti-β-actin (1:2,000, mouse monoclonal; Santa Cruz Biotechnology, Paso Robles, CA).

    Techniques: Activity Assay, MTT Assay

    Analysis of wild-type and mutant BSEP distribution by immunofluorescence. The resulting distribution of fluorescence was observed by laser confocal microscopy. Green represents BSEP and red represents calnexin (left) and Na/K-ATPase (right) and yellow represents co-localization of the two proteins. In each panel, an X-Y image (upper) and an X-Z image (lower) are shown. The data shown were obtained from four independent experiments. Scale bar, 5 µm.

    Journal: Molecular Medicine Reports

    Article Title: Functional analysis of the correlation between ABCB11 gene mutation and primary intrahepatic stone

    doi: 10.3892/mmr.2018.9661

    Figure Lengend Snippet: Analysis of wild-type and mutant BSEP distribution by immunofluorescence. The resulting distribution of fluorescence was observed by laser confocal microscopy. Green represents BSEP and red represents calnexin (left) and Na/K-ATPase (right) and yellow represents co-localization of the two proteins. In each panel, an X-Y image (upper) and an X-Z image (lower) are shown. The data shown were obtained from four independent experiments. Scale bar, 5 µm.

    Article Snippet: The primary antibodies used in this study included the following: Anti-BSEP (1:500; cat. no. 155421; Abcam, Cambridge, UK) and anti-Na/K-ATPase (1:1,000; cat. no. 58475; Abcam).

    Techniques: Mutagenesis, Immunofluorescence, Fluorescence, Confocal Microscopy

    (A) Structure of known proton-pump inhibitor, Rabeprazole (B) Gastric H+/K+-ATPase activity is inhibited by 100μM Rabeprazole. (C) Benzimidazole probes inhibit ATPase activity by approximately 50% at 10μM compound.

    Journal: Molecular bioSystems

    Article Title: Benzimidazole covalent probes and the gastric H+/K+-ATPase as a model system for protein labeling in a copper-free setting

    doi: 10.1039/c6mb00024j

    Figure Lengend Snippet: (A) Structure of known proton-pump inhibitor, Rabeprazole (B) Gastric H+/K+-ATPase activity is inhibited by 100μM Rabeprazole. (C) Benzimidazole probes inhibit ATPase activity by approximately 50% at 10μM compound.

    Article Snippet: Eluates were loaded onto a precast 4–15% Criterion™ Tris-HCl gel (BioRad) for protein separation, transferred to a PVDF membrane and blotted for the H,K-ATPase with Anti-ATP4A antibody (ab174293) (AbCam).

    Techniques: Activity Assay

    Benzimidazole probes label the gastric H+/K+-ATPase, ATP4A. (A) Schematic drawing of labeling protocol. After incubation of probe and gastric membranes biotin is “clicked” onto the probe, the target enriched using streptavidin beads, separated using SDS-PAGE and analyzed by western blot. (B) Labeling is pH dependent, requiring a pH of 3.0 for acid activation of benzimidazole compounds. (C) Direct comparison of labeling effi ciency between probes at 1μM shows that labeling is blockable with DTT (50mM). (D) Labeling with 0.3, 1, and 3μM probe shows that labeling is dose dependent and Rabe-Tz labels more efficiently than Rabe-N 3 at equivalent concentrations.

    Journal: Molecular bioSystems

    Article Title: Benzimidazole covalent probes and the gastric H+/K+-ATPase as a model system for protein labeling in a copper-free setting

    doi: 10.1039/c6mb00024j

    Figure Lengend Snippet: Benzimidazole probes label the gastric H+/K+-ATPase, ATP4A. (A) Schematic drawing of labeling protocol. After incubation of probe and gastric membranes biotin is “clicked” onto the probe, the target enriched using streptavidin beads, separated using SDS-PAGE and analyzed by western blot. (B) Labeling is pH dependent, requiring a pH of 3.0 for acid activation of benzimidazole compounds. (C) Direct comparison of labeling effi ciency between probes at 1μM shows that labeling is blockable with DTT (50mM). (D) Labeling with 0.3, 1, and 3μM probe shows that labeling is dose dependent and Rabe-Tz labels more efficiently than Rabe-N 3 at equivalent concentrations.

    Article Snippet: Eluates were loaded onto a precast 4–15% Criterion™ Tris-HCl gel (BioRad) for protein separation, transferred to a PVDF membrane and blotted for the H,K-ATPase with Anti-ATP4A antibody (ab174293) (AbCam).

    Techniques: Labeling, Incubation, SDS Page, Western Blot, Activation Assay

    Localization of ATP-dependent efflux transporter proteins. A-L ) Confocal micrographs after indirect immunofluorescence labeling with efflux pump proteins MRP-1, -4, or -5 (green), and eye-specific proteins MITF and cellular retinaldehyde-binding protein (CRALBP, both red), the polarization marker Na + /K + ATPase (red), and the nuclear label 4′,6′-diamidino-2-phenylidole (blue). In figures M-P ) the brightfield micrographs show the same ARPE-19 cells and fusiform, early cobblestone, and cobblestone hESC-RPE as shown in the confocal images. Scale bars, 10 µm.

    Journal: PLoS ONE

    Article Title: Efflux Protein Expression in Human Stem Cell-Derived Retinal Pigment Epithelial Cells

    doi: 10.1371/journal.pone.0030089

    Figure Lengend Snippet: Localization of ATP-dependent efflux transporter proteins. A-L ) Confocal micrographs after indirect immunofluorescence labeling with efflux pump proteins MRP-1, -4, or -5 (green), and eye-specific proteins MITF and cellular retinaldehyde-binding protein (CRALBP, both red), the polarization marker Na + /K + ATPase (red), and the nuclear label 4′,6′-diamidino-2-phenylidole (blue). In figures M-P ) the brightfield micrographs show the same ARPE-19 cells and fusiform, early cobblestone, and cobblestone hESC-RPE as shown in the confocal images. Scale bars, 10 µm.

    Article Snippet: Nonspecific binding sites were blocked with 3% BSA (Sigma-Aldrich) in PBS at RT for 1 h. Primary antibody incubations were done in 0.5% BSA-PBS, with rat monoclonal anti-MRP-1 (1∶100), anti-MRP-4 (1∶100), and anti-MRP-5 (1∶50), with rabbit anti-microphthalmia-associated transcription factor (MITF, 1∶350), mouse anti-cellular retinaldehyde-binding protein (CRALBP, 1∶1000), or mouse anti-Na+ /K+ ATPase (1∶50; all antibodies were from Abcam) for 1h.

    Techniques: Immunofluorescence, Labeling, Binding Assay, Marker