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  • 94
    Millipore hptlc plates
    Hptlc Plates, supplied by Millipore, used in various techniques. Bioz Stars score: 94/100, based on 51 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Camag camag hptlc system
    High-performance thin layer chromatography <t>(HPTLC)</t> images of the fingerprint of Valerianae Radix et Rhizoma water extract (VRe). VRe was loaded and developed in HPTLC plate, then detected with an ultraviolet or ((a) 254 nm; (b) 366 nm; (c) white light after p-anisaldehyde sprayed) valerenic acid standard. Images were obtained with <t>Camag</t> Visualizer (Camag, Swiss)
    Camag Hptlc System, supplied by Camag, used in various techniques. Bioz Stars score: 90/100, based on 420 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Merck KGaA hptlc plates
    F1394 increases free cholesterol/cholesterol ester ratio in LDs. <t>Huh7</t> cells were cultured for 24 h in the absence (Control) or presence of 10 μg/ml F1394 or 10 μg/ml 58-035 or treated with MβCD/cholesterol in LPDS (Chol) or MβCD/cholesterol plus 58-035 in LPDS (Chol + 58-035) as described in Materials and Methods . LD fractions were prepared and lipids analyzed by <t>HPTLC</t> as described in Materials and Methods . Results indicate the mean ± SD ( n = 3). * p = 0.05, ** p
    Hptlc Plates, supplied by Merck KGaA, used in various techniques. Bioz Stars score: 92/100, based on 213 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Camag hptlc plates
    The <t>HPTLC</t> plate developed with four different solvent systems, showing separated biosurfactant under UV 254 nm (A) , and the mass spectra of two bands directly extracted by <t>TLC–MS</t> interface and analyzed by ESI–MS (B) under positive mode.
    Hptlc Plates, supplied by Camag, used in various techniques. Bioz Stars score: 91/100, based on 190 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Merck KGaA silica gel 60 hptlc plates
    Lipid analysis of cortical sphingolipids. Lipids of the cortex were isolated from mice of the indicated ages and genotypes, subjected to mild alkaline methanolysis, and separated by TLC. A , The amount of lipids that corresponds to 0.5 mg (2.5 months) or 1 mg (5–6 months) of tissue wet weight was loaded per lane of silica <t>gel</t> 60 <t>HPTLC</t> plates, which was developed using chloroform/methanol/water (70:30:4). In CGT-tg mice, a lipid migrating between the NFA-GalCer (top band) and HFA-GalCer (bottom band) appeared (arrowhead). This lipid could be identified as C18:0-hexosylceramide and, because of its migration behavior in borate-treated TLC plates, as GalCer (data not shown). B , SGalCer was quantified by densitometry as described in Materials and Methods. Shown are the total SGalCer concentrations (total) and the concentrations of VLCFA-SGalCer (top band of the SGalCer douplet) and LCFA/HFA-containing SGalCer (bottom band). Data shown are mean ± SD [ n = 3; except for 2.5-month-old ASA(−/−) ( n = 6) and CGT/ASA(−/−) mice ( n = 4)]. Asterisks indicate a significant difference ( p
    Silica Gel 60 Hptlc Plates, supplied by Merck KGaA, used in various techniques. Bioz Stars score: 90/100, based on 126 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Merck & Co hptlc plates
    Cholesterol and ceramide-dependent partitioning of Sig-1R into DRMs. A, time-dependent depletion of free cholesterol caused by MβC. CHO cells in a serum-free medium were incubated with 5 mM MβC. Lipids were extracted, and free cholesterol was measured enzymatically. The graph represents the mean ± S.E.M. from five results. B, time-dependent depletion of free cholesterol caused by MβC. CHO cells treated with MβC for 15 or 60 min were fixed and stained with filipin. Bar, 50 μm. Results from five independent experiments are shown. C, depletion of <t>ceramides</t> at the MAM caused by FB1 (5 μg/ml, 4 h). Ceramides were extracted from MAM fractions. GM3 gangliosides were extracted from whole cells. Representative results from three independent <t>HPTLC</t> are shown. Lipids were visualized by ANS (top) and diphenylamine-aniline (bottom) sprays, respectively. D, PDMP (25 μg/ml, 4 h) decreases GlcCer but accumulates ceramides. Ceramides and GlcCer were extracted from whole cells. Representative results from three independent HPTLCs are shown. Lipids were visualized by ANS (top) and diphenylamine-aniline (bottom) sprays, respectively. E, effects of depletion of cholesterol or ceramide on partitioning of Src and Sig-1R to DRMs. Src-containing DRMs were extracted by 0.5% Tx whereas Sig-1R-containing DRMs were extracted by 0.5% Triton X-114. CHO cells were treated with MβC, FB1, or PDMP under the same conditions in B to D.
    Hptlc Plates, supplied by Merck & Co, used in various techniques. Bioz Stars score: 91/100, based on 86 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Merck KGaA high performance thin layer chromatography hptlc
    Comparative <t>HPTLC</t> profiles of standard, imperatorin (a); leaf extract (b) and <t>AME-2</t> (c).
    High Performance Thin Layer Chromatography Hptlc, supplied by Merck KGaA, used in various techniques. Bioz Stars score: 90/100, based on 82 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Merck & Co hptlc
    Comparative <t>HPTLC</t> profiles of standard, imperatorin (a); leaf extract (b) and <t>AME-2</t> (c).
    Hptlc, supplied by Merck & Co, used in various techniques. Bioz Stars score: 91/100, based on 129 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    Merck & Co silica gel 60 hptlc plates
    Lipid analysis of cortical sphingolipids. Lipids of the cortex were isolated from mice of the indicated ages and genotypes, subjected to mild alkaline methanolysis, and separated by TLC. A , The amount of lipids that corresponds to 0.5 mg (2.5 months) or 1 mg (5–6 months) of tissue wet weight was loaded per lane of silica <t>gel</t> 60 <t>HPTLC</t> plates, which was developed using chloroform/methanol/water (70:30:4). In CGT-tg mice, a lipid migrating between the NFA-GalCer (top band) and HFA-GalCer (bottom band) appeared (arrowhead). This lipid could be identified as C18:0-hexosylceramide and, because of its migration behavior in borate-treated TLC plates, as GalCer (data not shown). B , SGalCer was quantified by densitometry as described in Materials and Methods. Shown are the total SGalCer concentrations (total) and the concentrations of VLCFA-SGalCer (top band of the SGalCer douplet) and LCFA/HFA-containing SGalCer (bottom band). Data shown are mean ± SD [ n = 3; except for 2.5-month-old ASA(−/−) ( n = 6) and CGT/ASA(−/−) mice ( n = 4)]. Asterisks indicate a significant difference ( p
    Silica Gel 60 Hptlc Plates, supplied by Merck & Co, used in various techniques. Bioz Stars score: 89/100, based on 110 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Camag high performance thin layer chromatography hptlc
    Typical <t>HPTLC</t> densitogram of <t>mycophenolate</t> mofetil standard (Rf: 0.55 ± 0.02)
    High Performance Thin Layer Chromatography Hptlc, supplied by Camag, used in various techniques. Bioz Stars score: 89/100, based on 67 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Merck & Co hptlc silica gel 60
    Typical <t>HPTLC</t> densitogram of <t>mycophenolate</t> mofetil standard (Rf: 0.55 ± 0.02)
    Hptlc Silica Gel 60, supplied by Merck & Co, used in various techniques. Bioz Stars score: 90/100, based on 25 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Merck & Co silica 60 f254 hptlc plates
    Typical <t>HPTLC</t> densitogram of <t>mycophenolate</t> mofetil standard (Rf: 0.55 ± 0.02)
    Silica 60 F254 Hptlc Plates, supplied by Merck & Co, used in various techniques. Bioz Stars score: 88/100, based on 66 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Merck KGaA silica gel 60 f254 hptlc plates
    Typical <t>HPTLC</t> densitogram of <t>mycophenolate</t> mofetil standard (Rf: 0.55 ± 0.02)
    Silica Gel 60 F254 Hptlc Plates, supplied by Merck KGaA, used in various techniques. Bioz Stars score: 89/100, based on 68 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Camag camag hptlc instrumentation
    Typical <t>HPTLC</t> densitogram of <t>mycophenolate</t> mofetil standard (Rf: 0.55 ± 0.02)
    Camag Hptlc Instrumentation, supplied by Camag, used in various techniques. Bioz Stars score: 86/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore silica gel 60 hptlc plates
    Typical <t>HPTLC</t> densitogram of <t>mycophenolate</t> mofetil standard (Rf: 0.55 ± 0.02)
    Silica Gel 60 Hptlc Plates, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 20 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Merck KGaA high performance thin layer chromatography hptlc silica gel 60 plates
    Typical <t>HPTLC</t> densitogram of <t>mycophenolate</t> mofetil standard (Rf: 0.55 ± 0.02)
    High Performance Thin Layer Chromatography Hptlc Silica Gel 60 Plates, supplied by Merck KGaA, used in various techniques. Bioz Stars score: 89/100, based on 55 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    High-performance thin layer chromatography (HPTLC) images of the fingerprint of Valerianae Radix et Rhizoma water extract (VRe). VRe was loaded and developed in HPTLC plate, then detected with an ultraviolet or ((a) 254 nm; (b) 366 nm; (c) white light after p-anisaldehyde sprayed) valerenic acid standard. Images were obtained with Camag Visualizer (Camag, Swiss)

    Journal: Pharmacognosy Magazine

    Article Title: Effects of Valerianae Radix et Rhizoma extract on psychological stress in mice

    doi: 10.4103/0973-1296.153093

    Figure Lengend Snippet: High-performance thin layer chromatography (HPTLC) images of the fingerprint of Valerianae Radix et Rhizoma water extract (VRe). VRe was loaded and developed in HPTLC plate, then detected with an ultraviolet or ((a) 254 nm; (b) 366 nm; (c) white light after p-anisaldehyde sprayed) valerenic acid standard. Images were obtained with Camag Visualizer (Camag, Swiss)

    Article Snippet: VRe was loaded on the HPTLC plate beside valerenic acid (C15 H22 O2 ) standard, and the HPLC fingerprint image was detected with CAMAG HPTLC system with Wincat32 software [ ].

    Techniques: High Performance Thin Layer Chromatography

    HPTLC chromatogram of P. foetida methanol extract and standard β-sitosterol

    Journal: BMC Complementary and Alternative Medicine

    Article Title: Gastroprotective mechanism of Paederia foetida Linn. (Rubiaceae) – a popular edible plant used by the tribal community of North-East India

    doi: 10.1186/s12906-015-0831-0

    Figure Lengend Snippet: HPTLC chromatogram of P. foetida methanol extract and standard β-sitosterol

    Article Snippet: HPTLC analysis of methanol extract of P. foetida The methanol extract of P. foetida was evaluated by using high performance thin layer chromatography (HPTLC) system (Camag linomat 5, Switzerland) for qualitative estimation of β- sitosterol at 540 nm.

    Techniques: High Performance Thin Layer Chromatography

    F1394 increases free cholesterol/cholesterol ester ratio in LDs. Huh7 cells were cultured for 24 h in the absence (Control) or presence of 10 μg/ml F1394 or 10 μg/ml 58-035 or treated with MβCD/cholesterol in LPDS (Chol) or MβCD/cholesterol plus 58-035 in LPDS (Chol + 58-035) as described in Materials and Methods . LD fractions were prepared and lipids analyzed by HPTLC as described in Materials and Methods . Results indicate the mean ± SD ( n = 3). * p = 0.05, ** p

    Journal: Molecular Biology of the Cell

    Article Title: Acute accumulation of free cholesterol induces the degradation of perilipin 2 and Rab18-dependent fusion of ER and lipid droplets in cultured human hepatocytes

    doi: 10.1091/mbc.E15-10-0730

    Figure Lengend Snippet: F1394 increases free cholesterol/cholesterol ester ratio in LDs. Huh7 cells were cultured for 24 h in the absence (Control) or presence of 10 μg/ml F1394 or 10 μg/ml 58-035 or treated with MβCD/cholesterol in LPDS (Chol) or MβCD/cholesterol plus 58-035 in LPDS (Chol + 58-035) as described in Materials and Methods . LD fractions were prepared and lipids analyzed by HPTLC as described in Materials and Methods . Results indicate the mean ± SD ( n = 3). * p = 0.05, ** p

    Article Snippet: Thin-layer chromatography Lipids were extracted ( ) from Huh7 cells and applied to HPTLC plates (Merck, Darmstadt, Germany).

    Techniques: Cell Culture, High Performance Thin Layer Chromatography

    The HPTLC plate developed with four different solvent systems, showing separated biosurfactant under UV 254 nm (A) , and the mass spectra of two bands directly extracted by TLC–MS interface and analyzed by ESI–MS (B) under positive mode.

    Journal: Frontiers in Microbiology

    Article Title: Production, Characterization, and Application of Bacillus licheniformis W16 Biosurfactant in Enhancing Oil Recovery

    doi: 10.3389/fmicb.2016.01853

    Figure Lengend Snippet: The HPTLC plate developed with four different solvent systems, showing separated biosurfactant under UV 254 nm (A) , and the mass spectra of two bands directly extracted by TLC–MS interface and analyzed by ESI–MS (B) under positive mode.

    Article Snippet: Twenty microliter of biosurfactant (0.1 g l−1 ) sample was spotted onto a HPTLC plate (10 cm × 10 cm), Silica gel 60 – F254 (Merck, Germany).

    Techniques: High Performance Thin Layer Chromatography, Thin Layer Chromatography, Mass Spectrometry

    The HPTLC plate developed with four different solvent systems, showing separated biosurfactant under UV 254 nm (A) , and the mass spectra of two bands directly extracted by TLC–MS interface and analyzed by ESI–MS (B) under positive mode.

    Journal: Frontiers in Microbiology

    Article Title: Production, Characterization, and Application of Bacillus licheniformis W16 Biosurfactant in Enhancing Oil Recovery

    doi: 10.3389/fmicb.2016.01853

    Figure Lengend Snippet: The HPTLC plate developed with four different solvent systems, showing separated biosurfactant under UV 254 nm (A) , and the mass spectra of two bands directly extracted by TLC–MS interface and analyzed by ESI–MS (B) under positive mode.

    Article Snippet: The HPTLC plates were evaluated by TLC visualizer (CAMAG) under direct Ultra Violet (UV 254 nm) light, and images were captured.

    Techniques: High Performance Thin Layer Chromatography, Thin Layer Chromatography, Mass Spectrometry

    Lipid analysis of cortical sphingolipids. Lipids of the cortex were isolated from mice of the indicated ages and genotypes, subjected to mild alkaline methanolysis, and separated by TLC. A , The amount of lipids that corresponds to 0.5 mg (2.5 months) or 1 mg (5–6 months) of tissue wet weight was loaded per lane of silica gel 60 HPTLC plates, which was developed using chloroform/methanol/water (70:30:4). In CGT-tg mice, a lipid migrating between the NFA-GalCer (top band) and HFA-GalCer (bottom band) appeared (arrowhead). This lipid could be identified as C18:0-hexosylceramide and, because of its migration behavior in borate-treated TLC plates, as GalCer (data not shown). B , SGalCer was quantified by densitometry as described in Materials and Methods. Shown are the total SGalCer concentrations (total) and the concentrations of VLCFA-SGalCer (top band of the SGalCer douplet) and LCFA/HFA-containing SGalCer (bottom band). Data shown are mean ± SD [ n = 3; except for 2.5-month-old ASA(−/−) ( n = 6) and CGT/ASA(−/−) mice ( n = 4)]. Asterisks indicate a significant difference ( p

    Journal: The Journal of Neuroscience

    Article Title: Sulfatide Storage in Neurons Causes Hyperexcitability and Axonal Degeneration in a Mouse Model of Metachromatic Leukodystrophy

    doi: 10.1523/JNEUROSCI.2329-07.2007

    Figure Lengend Snippet: Lipid analysis of cortical sphingolipids. Lipids of the cortex were isolated from mice of the indicated ages and genotypes, subjected to mild alkaline methanolysis, and separated by TLC. A , The amount of lipids that corresponds to 0.5 mg (2.5 months) or 1 mg (5–6 months) of tissue wet weight was loaded per lane of silica gel 60 HPTLC plates, which was developed using chloroform/methanol/water (70:30:4). In CGT-tg mice, a lipid migrating between the NFA-GalCer (top band) and HFA-GalCer (bottom band) appeared (arrowhead). This lipid could be identified as C18:0-hexosylceramide and, because of its migration behavior in borate-treated TLC plates, as GalCer (data not shown). B , SGalCer was quantified by densitometry as described in Materials and Methods. Shown are the total SGalCer concentrations (total) and the concentrations of VLCFA-SGalCer (top band of the SGalCer douplet) and LCFA/HFA-containing SGalCer (bottom band). Data shown are mean ± SD [ n = 3; except for 2.5-month-old ASA(−/−) ( n = 6) and CGT/ASA(−/−) mice ( n = 4)]. Asterisks indicate a significant difference ( p

    Article Snippet: The aqueous phase was washed once with chloroform, and the combined organic phases were dried and separated by thin-layer chromatography (TLC) on silica gel 60 HPTLC plates (Merck, Darmstadt, Germany) using chloroform/methanol/water (70:30:4).

    Techniques: Isolation, Mouse Assay, Thin Layer Chromatography, High Performance Thin Layer Chromatography, Migration

    Cholesterol and ceramide-dependent partitioning of Sig-1R into DRMs. A, time-dependent depletion of free cholesterol caused by MβC. CHO cells in a serum-free medium were incubated with 5 mM MβC. Lipids were extracted, and free cholesterol was measured enzymatically. The graph represents the mean ± S.E.M. from five results. B, time-dependent depletion of free cholesterol caused by MβC. CHO cells treated with MβC for 15 or 60 min were fixed and stained with filipin. Bar, 50 μm. Results from five independent experiments are shown. C, depletion of ceramides at the MAM caused by FB1 (5 μg/ml, 4 h). Ceramides were extracted from MAM fractions. GM3 gangliosides were extracted from whole cells. Representative results from three independent HPTLC are shown. Lipids were visualized by ANS (top) and diphenylamine-aniline (bottom) sprays, respectively. D, PDMP (25 μg/ml, 4 h) decreases GlcCer but accumulates ceramides. Ceramides and GlcCer were extracted from whole cells. Representative results from three independent HPTLCs are shown. Lipids were visualized by ANS (top) and diphenylamine-aniline (bottom) sprays, respectively. E, effects of depletion of cholesterol or ceramide on partitioning of Src and Sig-1R to DRMs. Src-containing DRMs were extracted by 0.5% Tx whereas Sig-1R-containing DRMs were extracted by 0.5% Triton X-114. CHO cells were treated with MβC, FB1, or PDMP under the same conditions in B to D.

    Journal: Molecular Pharmacology

    Article Title: Detergent-Resistant Microdomains Determine the Localization of ?-1 Receptors to the Endoplasmic Reticulum-Mitochondria Junction S⃞

    doi: 10.1124/mol.109.062539

    Figure Lengend Snippet: Cholesterol and ceramide-dependent partitioning of Sig-1R into DRMs. A, time-dependent depletion of free cholesterol caused by MβC. CHO cells in a serum-free medium were incubated with 5 mM MβC. Lipids were extracted, and free cholesterol was measured enzymatically. The graph represents the mean ± S.E.M. from five results. B, time-dependent depletion of free cholesterol caused by MβC. CHO cells treated with MβC for 15 or 60 min were fixed and stained with filipin. Bar, 50 μm. Results from five independent experiments are shown. C, depletion of ceramides at the MAM caused by FB1 (5 μg/ml, 4 h). Ceramides were extracted from MAM fractions. GM3 gangliosides were extracted from whole cells. Representative results from three independent HPTLC are shown. Lipids were visualized by ANS (top) and diphenylamine-aniline (bottom) sprays, respectively. D, PDMP (25 μg/ml, 4 h) decreases GlcCer but accumulates ceramides. Ceramides and GlcCer were extracted from whole cells. Representative results from three independent HPTLCs are shown. Lipids were visualized by ANS (top) and diphenylamine-aniline (bottom) sprays, respectively. E, effects of depletion of cholesterol or ceramide on partitioning of Src and Sig-1R to DRMs. Src-containing DRMs were extracted by 0.5% Tx whereas Sig-1R-containing DRMs were extracted by 0.5% Triton X-114. CHO cells were treated with MβC, FB1, or PDMP under the same conditions in B to D.

    Article Snippet: After separation from other lipids, the ceramides were extracted from an HPTLC plate (Merck) and spotted on a Hybond-C membrane (GE Healthcare).

    Techniques: Incubation, Staining, High Performance Thin Layer Chromatography

    MAM accommodates DRMs associated with Sig-1R. A, purification of MAM and microsomal fractions from CHO cells. Nuclear (P1), mitochondrial (Mito), MAM, microsomal (P3), and cytosolic (Cyt) fractions were prepared by differential centrifugation combined with a Percoll gradient fractionation. Five micrograms of proteins was applied to SDS-PAGE, followed by immunoblotting. COX, cytochrome c oxidase subunit I. Asterisks indicate ER chaperones involved in vesicular transport. Graphs represent fraction distributions of proteins in which the sum of five fractions was taken as 100% for each protein. B, Sig-1R-associated DRMs in the MAM. MAM and microsomes (P3) (25 μg of total proteins in each) were extracted by 0.5% Tx (Tx-100) or Triton X-114 at 4°C. DRMs and detergent-soluble supernatant (S) were prepared by differential centrifugations. The numbers represent the average of optical density (O.D.) measured in each protein band ( n = 3). C, silver staining for total proteins associated with DRMs in MAM and microsomal fractions. MAM and microsomes (25 μg of total proteins in each) were extracted in 0.5% Tx at 4°C, and DRMs and soluble supernatants (S) were prepared. Proteins were visualized by 13% SDS-PAGE, followed by silver staining. The numbers represent the average of O.D. measured in each lane ( n = 3). MW, molecular weight of standard proteins. D, lipid contents in MAM and microsomal fractions. Lipids were extracted and analyzed by HPTLC. Cholesterol (Chol) was detected using a ferric chloride spray; GlcCer using a diphenylamine-aniline spray. Lipids in the second panel were visualized under UV light after an ANS spray. In the lipid overlay assay for ceramides (Cer, bottom), ceramides extracted from HPTLC plates were immobilized on a nitrocellulose membrane followed by immunoblotting with anti-ceramide antibodies. SM, sphingomyelin.

    Journal: Molecular Pharmacology

    Article Title: Detergent-Resistant Microdomains Determine the Localization of ?-1 Receptors to the Endoplasmic Reticulum-Mitochondria Junction S⃞

    doi: 10.1124/mol.109.062539

    Figure Lengend Snippet: MAM accommodates DRMs associated with Sig-1R. A, purification of MAM and microsomal fractions from CHO cells. Nuclear (P1), mitochondrial (Mito), MAM, microsomal (P3), and cytosolic (Cyt) fractions were prepared by differential centrifugation combined with a Percoll gradient fractionation. Five micrograms of proteins was applied to SDS-PAGE, followed by immunoblotting. COX, cytochrome c oxidase subunit I. Asterisks indicate ER chaperones involved in vesicular transport. Graphs represent fraction distributions of proteins in which the sum of five fractions was taken as 100% for each protein. B, Sig-1R-associated DRMs in the MAM. MAM and microsomes (P3) (25 μg of total proteins in each) were extracted by 0.5% Tx (Tx-100) or Triton X-114 at 4°C. DRMs and detergent-soluble supernatant (S) were prepared by differential centrifugations. The numbers represent the average of optical density (O.D.) measured in each protein band ( n = 3). C, silver staining for total proteins associated with DRMs in MAM and microsomal fractions. MAM and microsomes (25 μg of total proteins in each) were extracted in 0.5% Tx at 4°C, and DRMs and soluble supernatants (S) were prepared. Proteins were visualized by 13% SDS-PAGE, followed by silver staining. The numbers represent the average of O.D. measured in each lane ( n = 3). MW, molecular weight of standard proteins. D, lipid contents in MAM and microsomal fractions. Lipids were extracted and analyzed by HPTLC. Cholesterol (Chol) was detected using a ferric chloride spray; GlcCer using a diphenylamine-aniline spray. Lipids in the second panel were visualized under UV light after an ANS spray. In the lipid overlay assay for ceramides (Cer, bottom), ceramides extracted from HPTLC plates were immobilized on a nitrocellulose membrane followed by immunoblotting with anti-ceramide antibodies. SM, sphingomyelin.

    Article Snippet: After separation from other lipids, the ceramides were extracted from an HPTLC plate (Merck) and spotted on a Hybond-C membrane (GE Healthcare).

    Techniques: Purification, Centrifugation, Fractionation, SDS Page, Silver Staining, Molecular Weight, High Performance Thin Layer Chromatography, Overlay Assay

    MAM accumulates ceramides. A, accumulation of NBD-Cer at subcellular membranes. CHO cells were incubated with NBD-ceramide (1.25 μM) at 4°C for 30 min. Confocal microscope (left). Arrowhead, ER cisternae; small arrow, mitochondria with a spaghetti-like shape; large arrow, juxta-mitochondria structures accumulating NBD-Cer; N, nucleus. Bar, 10 μm. HPTLC (right), where lipid extracts from NBD-Cer-labeled CHO cells were visualized under UV. 1, the lipid extract from CHO cells incubated with NBD-Cer at 4°C for 30 min; 2, the lipid extract from CHO cells incubated with NBD-Cer at 4°C for 30 min, followed by washings and incubation at 37°C for 30 min. Note the few conversions of NBD-Cer to NBD-GlcCer in lane 1. B, accumulation of NBD-Cer at intracellular domains apposing mitochondria. NBD-Cer (top) and mitochondria expressing Mito-DsRed (bottom) were visualized in the same cell. The two panels were superimposed in the inset (NBD-Cer in green, MitoDsRed in red). Bar, 2.5 μm. C, MAM-specific distribution of Sig-1R (top; in green inset). Shown at the bottom is mitochondria in the same sample. Bar, 2.5 μm. Inset, two panels superimposed. D, colocalization between Sig-1R-EYFP and fluorescent ceramides. CHO cells expressing Sig-1R-EYFP (top) were labeled with fluorescence ceramide BODIPY-Cer TR (1.25 μM; bottom) at 4°C for 30 min. Inset, two panels are superimposed (Sig-1R-EYFP in green, BODIPY-Cer TR in red). Bar, 2.5 μm. Inset, colocalization is represented by yellow. The same results were obtained from four independent samples ( > 5 fields/sample). E, MAM accumulates NBD-Cer. CHO cells labeled with NBD-Cer at 4°C for 30 min were subjected to membrane fractionations. The level of NBD-Cer in each fraction was measured by a fluorescence microplate reader. Fluorescence intensities were normalized to protein concentrations in each sample. F, retention of NBD-Cer at MAM. CHO cells were labeled with NBD-ceramides at 4°C for 30 min (left image). After washings, cells were incubated in normal medium at 37°C for 30 min (right image). Thin arrows, mitochondria; thick arrows, MAM; N, nucleus; G, Golgi apparatus; Bar, 10 μm.

    Journal: Molecular Pharmacology

    Article Title: Detergent-Resistant Microdomains Determine the Localization of ?-1 Receptors to the Endoplasmic Reticulum-Mitochondria Junction S⃞

    doi: 10.1124/mol.109.062539

    Figure Lengend Snippet: MAM accumulates ceramides. A, accumulation of NBD-Cer at subcellular membranes. CHO cells were incubated with NBD-ceramide (1.25 μM) at 4°C for 30 min. Confocal microscope (left). Arrowhead, ER cisternae; small arrow, mitochondria with a spaghetti-like shape; large arrow, juxta-mitochondria structures accumulating NBD-Cer; N, nucleus. Bar, 10 μm. HPTLC (right), where lipid extracts from NBD-Cer-labeled CHO cells were visualized under UV. 1, the lipid extract from CHO cells incubated with NBD-Cer at 4°C for 30 min; 2, the lipid extract from CHO cells incubated with NBD-Cer at 4°C for 30 min, followed by washings and incubation at 37°C for 30 min. Note the few conversions of NBD-Cer to NBD-GlcCer in lane 1. B, accumulation of NBD-Cer at intracellular domains apposing mitochondria. NBD-Cer (top) and mitochondria expressing Mito-DsRed (bottom) were visualized in the same cell. The two panels were superimposed in the inset (NBD-Cer in green, MitoDsRed in red). Bar, 2.5 μm. C, MAM-specific distribution of Sig-1R (top; in green inset). Shown at the bottom is mitochondria in the same sample. Bar, 2.5 μm. Inset, two panels superimposed. D, colocalization between Sig-1R-EYFP and fluorescent ceramides. CHO cells expressing Sig-1R-EYFP (top) were labeled with fluorescence ceramide BODIPY-Cer TR (1.25 μM; bottom) at 4°C for 30 min. Inset, two panels are superimposed (Sig-1R-EYFP in green, BODIPY-Cer TR in red). Bar, 2.5 μm. Inset, colocalization is represented by yellow. The same results were obtained from four independent samples ( > 5 fields/sample). E, MAM accumulates NBD-Cer. CHO cells labeled with NBD-Cer at 4°C for 30 min were subjected to membrane fractionations. The level of NBD-Cer in each fraction was measured by a fluorescence microplate reader. Fluorescence intensities were normalized to protein concentrations in each sample. F, retention of NBD-Cer at MAM. CHO cells were labeled with NBD-ceramides at 4°C for 30 min (left image). After washings, cells were incubated in normal medium at 37°C for 30 min (right image). Thin arrows, mitochondria; thick arrows, MAM; N, nucleus; G, Golgi apparatus; Bar, 10 μm.

    Article Snippet: After separation from other lipids, the ceramides were extracted from an HPTLC plate (Merck) and spotted on a Hybond-C membrane (GE Healthcare).

    Techniques: Incubation, Microscopy, High Performance Thin Layer Chromatography, Labeling, Expressing, Fluorescence

    Comparative HPTLC profiles of standard, imperatorin (a); leaf extract (b) and AME-2 (c).

    Journal: Indian Journal of Pharmaceutical Sciences

    Article Title: Pharmacognostic standardisation and antiproliferative activity of Aegle marmelos (L.) Correa leaves in various human cancer cell lines

    doi:

    Figure Lengend Snippet: Comparative HPTLC profiles of standard, imperatorin (a); leaf extract (b) and AME-2 (c).

    Article Snippet: The most active fraction (AME-2) was subjected to further characterisation by high-performance thin layer chromatography (HPTLC) on precoated silica hexane:ethyl acetate plates (20×20, Merck, Germany) using Linomet-IV applicator and Camag TLC Scanner-III with WinCATS 4 software.

    Techniques: High Performance Thin Layer Chromatography

    Lipid analysis of cortical sphingolipids. Lipids of the cortex were isolated from mice of the indicated ages and genotypes, subjected to mild alkaline methanolysis, and separated by TLC. A , The amount of lipids that corresponds to 0.5 mg (2.5 months) or 1 mg (5–6 months) of tissue wet weight was loaded per lane of silica gel 60 HPTLC plates, which was developed using chloroform/methanol/water (70:30:4). In CGT-tg mice, a lipid migrating between the NFA-GalCer (top band) and HFA-GalCer (bottom band) appeared (arrowhead). This lipid could be identified as C18:0-hexosylceramide and, because of its migration behavior in borate-treated TLC plates, as GalCer (data not shown). B , SGalCer was quantified by densitometry as described in Materials and Methods. Shown are the total SGalCer concentrations (total) and the concentrations of VLCFA-SGalCer (top band of the SGalCer douplet) and LCFA/HFA-containing SGalCer (bottom band). Data shown are mean ± SD [ n = 3; except for 2.5-month-old ASA(−/−) ( n = 6) and CGT/ASA(−/−) mice ( n = 4)]. Asterisks indicate a significant difference ( p

    Journal: The Journal of Neuroscience

    Article Title: Sulfatide Storage in Neurons Causes Hyperexcitability and Axonal Degeneration in a Mouse Model of Metachromatic Leukodystrophy

    doi: 10.1523/JNEUROSCI.2329-07.2007

    Figure Lengend Snippet: Lipid analysis of cortical sphingolipids. Lipids of the cortex were isolated from mice of the indicated ages and genotypes, subjected to mild alkaline methanolysis, and separated by TLC. A , The amount of lipids that corresponds to 0.5 mg (2.5 months) or 1 mg (5–6 months) of tissue wet weight was loaded per lane of silica gel 60 HPTLC plates, which was developed using chloroform/methanol/water (70:30:4). In CGT-tg mice, a lipid migrating between the NFA-GalCer (top band) and HFA-GalCer (bottom band) appeared (arrowhead). This lipid could be identified as C18:0-hexosylceramide and, because of its migration behavior in borate-treated TLC plates, as GalCer (data not shown). B , SGalCer was quantified by densitometry as described in Materials and Methods. Shown are the total SGalCer concentrations (total) and the concentrations of VLCFA-SGalCer (top band of the SGalCer douplet) and LCFA/HFA-containing SGalCer (bottom band). Data shown are mean ± SD [ n = 3; except for 2.5-month-old ASA(−/−) ( n = 6) and CGT/ASA(−/−) mice ( n = 4)]. Asterisks indicate a significant difference ( p

    Article Snippet: Dried sphingolipids were dissolved in methanol/chloroform (1:1) and applied onto TLC or HPTLC silica gel 60 plates (Merck).

    Techniques: Isolation, Mouse Assay, Thin Layer Chromatography, High Performance Thin Layer Chromatography, Migration

    Typical HPTLC densitogram of mycophenolate mofetil standard (Rf: 0.55 ± 0.02)

    Journal: Pharmaceutical Methods

    Article Title: Development and validation of HPTLC method for the determination of mycophenolate mofetil in bulk and pharmaceutical formulation

    doi: 10.4103/2229-4708.103882

    Figure Lengend Snippet: Typical HPTLC densitogram of mycophenolate mofetil standard (Rf: 0.55 ± 0.02)

    Article Snippet: No high-performance thin layer chromatography (HPTLC) method for the quantitative analysis of mycophenolate mofetil as bulk and as formulation or for stability studies was found by a computer-assisted survey of the literature either from chemical abstracts or by using the CAMAG Bibliography Service.

    Techniques: High Performance Thin Layer Chromatography

    Typical HPTLC densitogram of mycophenolate mofetil extracted from tablet formulation (Rf: 0.55 ± 0.02)

    Journal: Pharmaceutical Methods

    Article Title: Development and validation of HPTLC method for the determination of mycophenolate mofetil in bulk and pharmaceutical formulation

    doi: 10.4103/2229-4708.103882

    Figure Lengend Snippet: Typical HPTLC densitogram of mycophenolate mofetil extracted from tablet formulation (Rf: 0.55 ± 0.02)

    Article Snippet: No high-performance thin layer chromatography (HPTLC) method for the quantitative analysis of mycophenolate mofetil as bulk and as formulation or for stability studies was found by a computer-assisted survey of the literature either from chemical abstracts or by using the CAMAG Bibliography Service.

    Techniques: High Performance Thin Layer Chromatography