lc ms buffer a  (Millipore)


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    Structured Review

    Millipore lc ms buffer a
    Lc Ms Buffer A, supplied by Millipore, 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/lc ms buffer a/product/Millipore
    Average 91 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    lc ms buffer a - by Bioz Stars, 2020-09
    91/100 stars

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    Liquid Chromatography with Mass Spectroscopy:

    Article Title: Purification and characterisation of dsRNA using ion pair reverse phase chromatography and mass spectrometry
    Article Snippet: .. LC MS buffer A: 80 mM 1,1,1,3,3,3,-Hexafluoro-2-propanol (HFIP, Sigma-Aldrich) with 20 mM TEAA (Sigma-Aldrich), LC MS buffer B: LC MS buffer A with 50% acetonitrile (v/v) (ThermoFisher). .. Starting with 10% buffer B, an isocratic step was performed for 2 mins followed by a linear extension to 20% B in 20 mins, then 20%B to 30% B over 10 min and 30%B to 90%B for 3 mins at a flow rate of 100 μL min−1 .

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    Millipore gabapentin
    Altered hindpaw gait indices in mice with neuropathy are not influenced by the analgesic <t>gabapentin.</t> ( A ) Systemic administration (i.p.) of gabapentin (30 mg/kg) did not significantly attenuate altered hindpaw gait indices in SNI mice. Mice were tested 1 h post saline/gabapentin administration. Data are plotted as the ratio of individual gait parameters in ipsilateral (right) vs contralateral (left) hindpaws; mean ± SEM (n=8 per group). ** denotes p
    Gabapentin, supplied by Millipore, used in various techniques. Bioz Stars score: 96/100, based on 21 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 96 stars, based on 21 article reviews
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    89
    Millipore microcon ultrafiltration uf membranes
    Enhancements to the FASP workflow. Samples are prepared in 4% SDS and diluted with 8 M urea to dissociate SDS from proteins. <t>Microcon</t> filter units and collection tubes are passivated by overnight incubation with 5% TWEEN-20, followed by thorough washing in MS-grade water. Diluted samples are then applied to passivated Microcon filters for buffer exchanges, eliminating SDS and contaminants. Proteins are alkylated with urea present, followed by successive buffer exchanges. Proteins are digested in the presence of surfactants, with the product peptides liberated by centrifugation. Extracting surfactant into an organic layer leaves behind pure peptides for fractionation or directly analysis by LC–MS.
    Microcon Ultrafiltration Uf Membranes, supplied by Millipore, used in various techniques. Bioz Stars score: 89/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/microcon ultrafiltration uf membranes/product/Millipore
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    91
    Millipore fura 2 loading buffer
    Bioassay-linked fractionation of TRPM2-active organic extract of Cacospongia sp. (A, B) Hydrogen peroxide-induced Ca 2+ entry in our calcium imaging assay using TRPM2-HEK293 cells following (A) treatment with vehicle control (closed circles, n = 15) versus an organic extract of Cacospongia sp. (open circles, n = 3) or (B)treatment with vehicle control (closed circles, n = 20) versus HPLC fractions of the extract eluting at 12.0–12.5 min (black trace, open circles, n = 2), 12.5–13.0 min (red trace, open circles, n = 2), 13.0–13.5 min (red trace, close circles, n = 2), or 13.5–14.0 min (black trace, closed squares, n = 2). H 2 O 2 (250 μ M) was added at 30 s. Fractions (approximately 17 μ g/mL each) were incubated for 20–30 min. Cells were preincubated with <t>fura-2</t> AM, and change in relative fluorescence units was measured at 510 nm after excitation at 340 nm.(C)Chromatogram of semipreparative reversed-phase HPLC of the organic extract residue. Eluent (2.0–30.0 min) was collected into 96-well plates in 30–60 s fractions and tested at proportional concentrations in the TRPM2 Ca 2+ following treatment with vehicle control (closed circles, n = 4) versus HPLC fractions of the Cacospongia sp. extract eluting at 13.0–14.0 min (approximately 1 μ g/mL, open circles, n = 4). Cells were incubated with the indicated fraction/control for 30–60 min in standard external Ringer’s solution before patching. Internal solution was unbuffered (no Ca 2+ chelator) standard K-glutamate-based solution supplemented with 100 μ M ADPR. Currents were elicited by voltage ramps from −100 mV to 100 mV over 50 ms at 0.5 Hz intervals. Current amplitudes were extracted at −80 mV, normalized to the current assessed at 100 s, averaged, and plotted versus time of the experiment. Induction time for tetracycline was 6–9 h. Error bars indicate SEM. For panels A, B, and D, data at the 100 s time point were statistically evaluated. Each of the following samples showed significance compared to control: (A) crude extract; (B) 12.5–13.0 and 13.0–13.5 min fractions; (D) 13.0–14.0 min fraction.
    Fura 2 Loading Buffer, supplied by Millipore, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Altered hindpaw gait indices in mice with neuropathy are not influenced by the analgesic gabapentin. ( A ) Systemic administration (i.p.) of gabapentin (30 mg/kg) did not significantly attenuate altered hindpaw gait indices in SNI mice. Mice were tested 1 h post saline/gabapentin administration. Data are plotted as the ratio of individual gait parameters in ipsilateral (right) vs contralateral (left) hindpaws; mean ± SEM (n=8 per group). ** denotes p

    Journal: Neuropharmacology

    Article Title: Pharmacological Validation of Voluntary Gait and Mechanical Sensitivity Assays Associated with Inflammatory and Neuropathic Pain in Mice

    doi: 10.1016/j.neuropharm.2017.11.036

    Figure Lengend Snippet: Altered hindpaw gait indices in mice with neuropathy are not influenced by the analgesic gabapentin. ( A ) Systemic administration (i.p.) of gabapentin (30 mg/kg) did not significantly attenuate altered hindpaw gait indices in SNI mice. Mice were tested 1 h post saline/gabapentin administration. Data are plotted as the ratio of individual gait parameters in ipsilateral (right) vs contralateral (left) hindpaws; mean ± SEM (n=8 per group). ** denotes p

    Article Snippet: Gabapentin (MilliporeSigma, St. Louis, MO) was administered to sham and SNI surgery animals at 30 mg/kg i.p. 1 hour prior to testing on post-operative days 11 and 20, and carprofen (Zoetis, Inc., Kalamazoo, MI) was administered Saline and CFA-injected animals at 5 mg/kg i.p., starting 1 hour prior to testing on days 2, 4 and 5 following saline/CFA injection.

    Techniques: Mouse Assay

    Hindpaw inflammation and peripheral nerve injury augment avoidance in a mechanical conflict-avoidance test, which could be attenuated by known analgesic drugs. ( A ) Cartoon depicting the three-chamber mechanical conflict-avoidance test system, used as a non-reflexive, voluntary, pain-indicating behavioral assay in mice. ( B ) Hindpaw injection of CFA (10 μl of a 1 mg/ml suspension) into mouse hindpaws significantly increased the escape latency from the white-lit chamber compared to baseline, with a 5 mm probe height in the conflict chamber, tested at 4 days (4D) post-saline/CFA injection. Systemic administration (i.p.) of carprofen (10 mg/kg) significantly attenuated this CFA-induced increase in escape latency from the white-lit chamber, with a 5 mm probe height in the conflict chamber. Mice were tested 1.5 h post saline/carprofen administration on 4D. Data are plotted as mean ± SEM (with individual latencies in overlay) of escape latency from white-lit chamber with increasing height of mechanical probes in the conflict chamber (n=7 per group). ( C ) Compared to baseline, induction of spared nerve injury (SNI) significantly increased the escape latency from the white-lit chamber, with a 5 mm probe height in the conflict chamber, tested at 8 days (8D) post-sham/SNI surgery. Systemic administration (i.p.) of buprenorphine (25 μg/kg) significantly attenuated the increase in escape latency from the white-lit chamber, with a 5 mm probe height in the conflict chamber. Mice were tested 1.5 h post saline/ buprenorphine administration on 8D. Data are plotted in the same format as panel B (Sham+Buprenorphine n=6; SNI+Saline/SNI+Buprenorphine n=7/group). ( D ) In mice tested only at 20 days post-sham/SNI surgery, SNI induced an increase in the escape latency from the white-lit chamber, with a 5 mm probe height in the conflict chamber, an increase which could be reversed by systemic administration (i.p.) of gabapentin (30 mg/kg). Mice were tested 1.5 h post saline/gabapentin administration. Data are plotted in the same format as panels B and C (n=8 per group). In panels B–D , *** denotes p

    Journal: Neuropharmacology

    Article Title: Pharmacological Validation of Voluntary Gait and Mechanical Sensitivity Assays Associated with Inflammatory and Neuropathic Pain in Mice

    doi: 10.1016/j.neuropharm.2017.11.036

    Figure Lengend Snippet: Hindpaw inflammation and peripheral nerve injury augment avoidance in a mechanical conflict-avoidance test, which could be attenuated by known analgesic drugs. ( A ) Cartoon depicting the three-chamber mechanical conflict-avoidance test system, used as a non-reflexive, voluntary, pain-indicating behavioral assay in mice. ( B ) Hindpaw injection of CFA (10 μl of a 1 mg/ml suspension) into mouse hindpaws significantly increased the escape latency from the white-lit chamber compared to baseline, with a 5 mm probe height in the conflict chamber, tested at 4 days (4D) post-saline/CFA injection. Systemic administration (i.p.) of carprofen (10 mg/kg) significantly attenuated this CFA-induced increase in escape latency from the white-lit chamber, with a 5 mm probe height in the conflict chamber. Mice were tested 1.5 h post saline/carprofen administration on 4D. Data are plotted as mean ± SEM (with individual latencies in overlay) of escape latency from white-lit chamber with increasing height of mechanical probes in the conflict chamber (n=7 per group). ( C ) Compared to baseline, induction of spared nerve injury (SNI) significantly increased the escape latency from the white-lit chamber, with a 5 mm probe height in the conflict chamber, tested at 8 days (8D) post-sham/SNI surgery. Systemic administration (i.p.) of buprenorphine (25 μg/kg) significantly attenuated the increase in escape latency from the white-lit chamber, with a 5 mm probe height in the conflict chamber. Mice were tested 1.5 h post saline/ buprenorphine administration on 8D. Data are plotted in the same format as panel B (Sham+Buprenorphine n=6; SNI+Saline/SNI+Buprenorphine n=7/group). ( D ) In mice tested only at 20 days post-sham/SNI surgery, SNI induced an increase in the escape latency from the white-lit chamber, with a 5 mm probe height in the conflict chamber, an increase which could be reversed by systemic administration (i.p.) of gabapentin (30 mg/kg). Mice were tested 1.5 h post saline/gabapentin administration. Data are plotted in the same format as panels B and C (n=8 per group). In panels B–D , *** denotes p

    Article Snippet: Gabapentin (MilliporeSigma, St. Louis, MO) was administered to sham and SNI surgery animals at 30 mg/kg i.p. 1 hour prior to testing on post-operative days 11 and 20, and carprofen (Zoetis, Inc., Kalamazoo, MI) was administered Saline and CFA-injected animals at 5 mg/kg i.p., starting 1 hour prior to testing on days 2, 4 and 5 following saline/CFA injection.

    Techniques: Behavioral Assay, Mouse Assay, Injection

    Enhancements to the FASP workflow. Samples are prepared in 4% SDS and diluted with 8 M urea to dissociate SDS from proteins. Microcon filter units and collection tubes are passivated by overnight incubation with 5% TWEEN-20, followed by thorough washing in MS-grade water. Diluted samples are then applied to passivated Microcon filters for buffer exchanges, eliminating SDS and contaminants. Proteins are alkylated with urea present, followed by successive buffer exchanges. Proteins are digested in the presence of surfactants, with the product peptides liberated by centrifugation. Extracting surfactant into an organic layer leaves behind pure peptides for fractionation or directly analysis by LC–MS.

    Journal: Journal of Proteome Research

    Article Title: Enhanced FASP (eFASP) to Increase Proteome Coverage and Sample Recovery for Quantitative Proteomic Experiments

    doi: 10.1021/pr4010019

    Figure Lengend Snippet: Enhancements to the FASP workflow. Samples are prepared in 4% SDS and diluted with 8 M urea to dissociate SDS from proteins. Microcon filter units and collection tubes are passivated by overnight incubation with 5% TWEEN-20, followed by thorough washing in MS-grade water. Diluted samples are then applied to passivated Microcon filters for buffer exchanges, eliminating SDS and contaminants. Proteins are alkylated with urea present, followed by successive buffer exchanges. Proteins are digested in the presence of surfactants, with the product peptides liberated by centrifugation. Extracting surfactant into an organic layer leaves behind pure peptides for fractionation or directly analysis by LC–MS.

    Article Snippet: Second, proteins remain denatured in urea, permitting Microcon ultrafiltration (UF) membranes with pore sizes up to 30 kDa to be employed, significantly reducing the time required for buffer exchange., One concern for the future use of FASP was the discontinuation of the Millipore Microcon centrifugal filters.

    Techniques: Incubation, Mass Spectrometry, Centrifugation, Fractionation, Liquid Chromatography with Mass Spectroscopy

    Bioassay-linked fractionation of TRPM2-active organic extract of Cacospongia sp. (A, B) Hydrogen peroxide-induced Ca 2+ entry in our calcium imaging assay using TRPM2-HEK293 cells following (A) treatment with vehicle control (closed circles, n = 15) versus an organic extract of Cacospongia sp. (open circles, n = 3) or (B)treatment with vehicle control (closed circles, n = 20) versus HPLC fractions of the extract eluting at 12.0–12.5 min (black trace, open circles, n = 2), 12.5–13.0 min (red trace, open circles, n = 2), 13.0–13.5 min (red trace, close circles, n = 2), or 13.5–14.0 min (black trace, closed squares, n = 2). H 2 O 2 (250 μ M) was added at 30 s. Fractions (approximately 17 μ g/mL each) were incubated for 20–30 min. Cells were preincubated with fura-2 AM, and change in relative fluorescence units was measured at 510 nm after excitation at 340 nm.(C)Chromatogram of semipreparative reversed-phase HPLC of the organic extract residue. Eluent (2.0–30.0 min) was collected into 96-well plates in 30–60 s fractions and tested at proportional concentrations in the TRPM2 Ca 2+ following treatment with vehicle control (closed circles, n = 4) versus HPLC fractions of the Cacospongia sp. extract eluting at 13.0–14.0 min (approximately 1 μ g/mL, open circles, n = 4). Cells were incubated with the indicated fraction/control for 30–60 min in standard external Ringer’s solution before patching. Internal solution was unbuffered (no Ca 2+ chelator) standard K-glutamate-based solution supplemented with 100 μ M ADPR. Currents were elicited by voltage ramps from −100 mV to 100 mV over 50 ms at 0.5 Hz intervals. Current amplitudes were extracted at −80 mV, normalized to the current assessed at 100 s, averaged, and plotted versus time of the experiment. Induction time for tetracycline was 6–9 h. Error bars indicate SEM. For panels A, B, and D, data at the 100 s time point were statistically evaluated. Each of the following samples showed significance compared to control: (A) crude extract; (B) 12.5–13.0 and 13.0–13.5 min fractions; (D) 13.0–14.0 min fraction.

    Journal: Journal of natural products

    Article Title: Scalaradial Is a Potent Inhibitor of Transient Receptor Potential Melastatin 2 (TRPM2) Ion Channels

    doi: 10.1021/acs.jnatprod.7b00515

    Figure Lengend Snippet: Bioassay-linked fractionation of TRPM2-active organic extract of Cacospongia sp. (A, B) Hydrogen peroxide-induced Ca 2+ entry in our calcium imaging assay using TRPM2-HEK293 cells following (A) treatment with vehicle control (closed circles, n = 15) versus an organic extract of Cacospongia sp. (open circles, n = 3) or (B)treatment with vehicle control (closed circles, n = 20) versus HPLC fractions of the extract eluting at 12.0–12.5 min (black trace, open circles, n = 2), 12.5–13.0 min (red trace, open circles, n = 2), 13.0–13.5 min (red trace, close circles, n = 2), or 13.5–14.0 min (black trace, closed squares, n = 2). H 2 O 2 (250 μ M) was added at 30 s. Fractions (approximately 17 μ g/mL each) were incubated for 20–30 min. Cells were preincubated with fura-2 AM, and change in relative fluorescence units was measured at 510 nm after excitation at 340 nm.(C)Chromatogram of semipreparative reversed-phase HPLC of the organic extract residue. Eluent (2.0–30.0 min) was collected into 96-well plates in 30–60 s fractions and tested at proportional concentrations in the TRPM2 Ca 2+ following treatment with vehicle control (closed circles, n = 4) versus HPLC fractions of the Cacospongia sp. extract eluting at 13.0–14.0 min (approximately 1 μ g/mL, open circles, n = 4). Cells were incubated with the indicated fraction/control for 30–60 min in standard external Ringer’s solution before patching. Internal solution was unbuffered (no Ca 2+ chelator) standard K-glutamate-based solution supplemented with 100 μ M ADPR. Currents were elicited by voltage ramps from −100 mV to 100 mV over 50 ms at 0.5 Hz intervals. Current amplitudes were extracted at −80 mV, normalized to the current assessed at 100 s, averaged, and plotted versus time of the experiment. Induction time for tetracycline was 6–9 h. Error bars indicate SEM. For panels A, B, and D, data at the 100 s time point were statistically evaluated. Each of the following samples showed significance compared to control: (A) crude extract; (B) 12.5–13.0 and 13.0–13.5 min fractions; (D) 13.0–14.0 min fraction.

    Article Snippet: The culture medium was completely removed at 16–20 h post induction and replaced with the fura-2 loading buffer: KRH buffer supplemented with 2 μ M fura-2 acetoxymethyl ester (fura-2 AM; Calbiochem) and 0.1% Pluronic F-127.

    Techniques: Fractionation, Imaging, High Performance Liquid Chromatography, Incubation, Fluorescence, Mass Spectrometry

    Effect of PARP-1, sPLA 2 , and PI3K/Akt pathway inhibitors on TRPM2. (A) In a calcium imaging assay, TRPM2-HEK cells were plated and induced for 20 h. Following fura-2 AM loading, the PARP-1 inhibitors PJ34 and DPQ were added. Cells were stimulated with 1.0 mM H 2 O 2 30 s after addition of the PARP-1 inhibitors, and the changes in intracellular [Ca 2+ ] were followed for 120 s. Change of the Ca 2+ signal is expressed as the maximum ratio change of fluorescence (at 510 nm, after excitation at 340/380 nm) normalized to vehicle as control (=100%). Each data point is the mean (±SEM) of quadruplicate determinations. Data are from one representative experiment. (B) In patch clamp experiments, TRPM2-HEK293 cells were incubated with the indicated concentrations of PARP-1 inhibitors in standard external Ringer’s solution, and whole-cell currents were recorded during the first 60 min. Internal solution was standard K-glutamate solution with 100 μ M ADPR. Current amplitudes were extracted at −80 mV and normalized to percent of controls recorded on the same day. Bar graphs show means of n = 3–17. Error bars are SEM. (C) In patch clamp experiments, TRPM2-HEK293 cells were incubated with the indicated concentrations of sPLA 2 inhibitor in standard external Ringer’s solution, and whole-cell currents were recorded during the first 60 min. Internal solution was standard K-glutamate solution with 100 μ M ADPR. Current amplitudes were extracted at −80 mV and normalized to percent of controls recorded on the same day. Bar graphs show means of n = 3–17. Error bars are SEM. (D) In patch clamp experiments, TRPM2-HEK293 cells were preincubated for 60 min with the indicated concentrations of Akt and PI3K inhibitor in serum-containing medium, before being transferred into standard external Ringer’s solution. Currents were recorded in the continued presence of the inhibitors. Internal solution was standard K-glutamate solution supplemented with 500 μ M ADPR. Current amplitudes were extracted at −80 mV and normalized to percent of controls recorded on the same day. Bar graphs show means of n = 10–19. Error bars are SEM.

    Journal: Journal of natural products

    Article Title: Scalaradial Is a Potent Inhibitor of Transient Receptor Potential Melastatin 2 (TRPM2) Ion Channels

    doi: 10.1021/acs.jnatprod.7b00515

    Figure Lengend Snippet: Effect of PARP-1, sPLA 2 , and PI3K/Akt pathway inhibitors on TRPM2. (A) In a calcium imaging assay, TRPM2-HEK cells were plated and induced for 20 h. Following fura-2 AM loading, the PARP-1 inhibitors PJ34 and DPQ were added. Cells were stimulated with 1.0 mM H 2 O 2 30 s after addition of the PARP-1 inhibitors, and the changes in intracellular [Ca 2+ ] were followed for 120 s. Change of the Ca 2+ signal is expressed as the maximum ratio change of fluorescence (at 510 nm, after excitation at 340/380 nm) normalized to vehicle as control (=100%). Each data point is the mean (±SEM) of quadruplicate determinations. Data are from one representative experiment. (B) In patch clamp experiments, TRPM2-HEK293 cells were incubated with the indicated concentrations of PARP-1 inhibitors in standard external Ringer’s solution, and whole-cell currents were recorded during the first 60 min. Internal solution was standard K-glutamate solution with 100 μ M ADPR. Current amplitudes were extracted at −80 mV and normalized to percent of controls recorded on the same day. Bar graphs show means of n = 3–17. Error bars are SEM. (C) In patch clamp experiments, TRPM2-HEK293 cells were incubated with the indicated concentrations of sPLA 2 inhibitor in standard external Ringer’s solution, and whole-cell currents were recorded during the first 60 min. Internal solution was standard K-glutamate solution with 100 μ M ADPR. Current amplitudes were extracted at −80 mV and normalized to percent of controls recorded on the same day. Bar graphs show means of n = 3–17. Error bars are SEM. (D) In patch clamp experiments, TRPM2-HEK293 cells were preincubated for 60 min with the indicated concentrations of Akt and PI3K inhibitor in serum-containing medium, before being transferred into standard external Ringer’s solution. Currents were recorded in the continued presence of the inhibitors. Internal solution was standard K-glutamate solution supplemented with 500 μ M ADPR. Current amplitudes were extracted at −80 mV and normalized to percent of controls recorded on the same day. Bar graphs show means of n = 10–19. Error bars are SEM.

    Article Snippet: The culture medium was completely removed at 16–20 h post induction and replaced with the fura-2 loading buffer: KRH buffer supplemented with 2 μ M fura-2 acetoxymethyl ester (fura-2 AM; Calbiochem) and 0.1% Pluronic F-127.

    Techniques: Imaging, Fluorescence, Patch Clamp, Incubation