Structured Review

Promega sequencing grade modified trypsin
Sequencing Grade Modified Trypsin, supplied by Promega, used in various techniques. Bioz Stars score: 99/100, based on 43 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/sequencing grade modified trypsin/product/Promega
Average 99 stars, based on 43 article reviews
Price from $9.99 to $1999.99
sequencing grade modified trypsin - by Bioz Stars, 2020-04
99/100 stars

Images

Related Articles

Expressing:

Article Title: The key protein of endosomal mRNP transport Rrm4 binds translational landmark sites of cargo mRNAs
Article Snippet: Preliminary purification experiments were performed using a strain expressing Rrm4 fused C‐terminally to Gfp and tandem affinity purification (TAP) tag . .. Samples were digested with sequencing‐grade modified trypsin (Promega), and the resulting peptides were separated into fractions by nanoLC (PepMap100 C‐18 RP nanocolumn and UltiMate 3000 liquid chromatography system, Dionex).

Blocking Assay:

Article Title: Profiling constitutive proteolytic events in vivo
Article Snippet: 1 h. This NHS-reactive reagent is specific for the N-terminal of the proteins, i.e. native N-termini and proteolytic cleavage sites, owing to the previous blocking of cysteine and lysine side chains, and the biotin tag of the molecule allows for positive selection by immobilized streptavidin. .. The samples were diluted 1:2 with distilled water before digestion overnight by sequencing-grade modified trypsin (Promega) or endoproteinase Glu-C (Roche).

Article Title: Sensitive Determination of Proteolytic Proteoforms in Limited Microscale Proteome Samples *
Article Snippet: Dimethyl labeling was used to block amine groups within the proteome. .. Subsequently, trypsin (1 mg/ml, Promega, cat. no. V5113) was introduced and incubated at 37 °C overnight.

Electrophoresis:

Article Title: Cross-Species Analysis of Nicotine-Induced Proteomic Alterations in Pancreatic Cells
Article Snippet: SeeBluePlus2 Pre-Stained standard (LC5925), LDS (lithium dodecyl sulfate) sample buffer (NP0008), NuPAGE 4-12% Bis-Tris polyacrylamide gels (NP0335), SimplyBlue Coomassie stain (LC0665), and MES-SDS (2-(N-morpholino) ethanesulfonic acid-sodium dodecyl sulfate) electrophoresis buffer (NP002) were from Invitrogen (Carlsbad, CA). (-)-Nicotine (≥99%) (N3876) was purchased from Sigma (St. Louis, MO). .. Sequencing-grade modified trypsin (V5111) was obtained from Promega (Madison, WI).

Incubation:

Article Title: The internal interaction in RBBP5 regulates assembly and activity of MLL1 methyltransferase complex
Article Snippet: Then Sulfo-NHS (N -hydroxysulfosuccinimide) was added to the reaction mixture at a final concentration of 2 mM, followed by incubation at room temperature for 2 h. Finally, the reaction was quenched by adding 20 mM Tris (pH 8.0). .. Trypsin digestion was performed with sequencing-grade modified trypsin (Promega, USA) at 37°C overnight.

Article Title: Sensitive Determination of Proteolytic Proteoforms in Limited Microscale Proteome Samples *
Article Snippet: .. Subsequently, trypsin (1 mg/ml, Promega, cat. no. V5113) was introduced and incubated at 37 °C overnight. ..

Isotopic Labeling:

Article Title: Sensitive Determination of Proteolytic Proteoforms in Limited Microscale Proteome Samples *
Article Snippet: Terminal amine isotopic labeling of substrates was performed according to the detailed terminal amine isotopic labeling of substrates (TAILS) procedure as described in the official TAILS bench protocol v4 by the Overall Lab ( http://clip.ubc.ca/resources/protocols-and-sops/ , May 2016). .. Subsequently, trypsin (1 mg/ml, Promega, cat. no. V5113) was introduced and incubated at 37 °C overnight.

Activity Assay:

Article Title: Profiling constitutive proteolytic events in vivo
Article Snippet: The samples were diluted 1:2 with distilled water before digestion overnight by sequencing-grade modified trypsin (Promega) or endoproteinase Glu-C (Roche). .. Further enzymatic activity was inhibited by boiling the samples for 5 min.

Mass Spectrometry:

Article Title: A proteomic-based approach to study underlying molecular responses of the small intestine of Wistar rats to genetically modified corn (MON810)
Article Snippet: Liquid chromatography–mass spectrometry (LC–MS/MS) and database search Selected protein spots were excised manually from the gel, and digested with sequencing grade-modified trypsin (Promega, USA) as described by Shevchenko et al. ( ). .. An ion-trap mass spectrometer (amaZon ETD, Bruker Daltonics, Bremen, Germany) equipped with an on-line nanospray source was used for mass spectrometry data acquisition.

Article Title: Methionine oxidation impairs reverse cholesterol transport by apolipoprotein A-I
Article Snippet: HDL was digested with sequencing grade modified trypsin (Promega) or sequencing grade endoproteinase Glu-C (Roche Applied Science) ( , , ). .. MS and MS/MS analyses were performed in the positive ion mode (mass range 200–2,000 Da) with a Thermo-Finnigan LCQ Deca XP Plus instrument ( , , ).

Article Title: The internal interaction in RBBP5 regulates assembly and activity of MLL1 methyltransferase complex
Article Snippet: Paragraph title: Cross-linking mass spectrometry ... Trypsin digestion was performed with sequencing-grade modified trypsin (Promega, USA) at 37°C overnight.

Article Title: Top-Down ETD-MS Provides Unreliable Quantitation of Methionine Oxidation
Article Snippet: Sequencing-grade modified trypsin was purchased from Promega (Madison, WI, USA). .. Sequencing-grade modified trypsin was purchased from Promega (Madison, WI, USA).

Article Title: Sensitive Determination of Proteolytic Proteoforms in Limited Microscale Proteome Samples *
Article Snippet: All the fractions were dried in a SpeedVac centrifuge and resuspended in 0.1% FA in water (Thermo Scientific, cat. no. SC2352911) prior to mass spectrometry analysis. .. Subsequently, trypsin (1 mg/ml, Promega, cat. no. V5113) was introduced and incubated at 37 °C overnight.

Article Title: The key protein of endosomal mRNP transport Rrm4 binds translational landmark sites of cargo mRNAs
Article Snippet: We analysed the eluted fractions by mass spectrometry either as complex mixture in solution (Fig C) or by cutting individual bands from an SDS–PAGE (Fig D). .. Samples were digested with sequencing‐grade modified trypsin (Promega), and the resulting peptides were separated into fractions by nanoLC (PepMap100 C‐18 RP nanocolumn and UltiMate 3000 liquid chromatography system, Dionex).

Modification:

Article Title: Microfluidic LC Device with Orthogonal Sample Extraction for On-Chip MALDI-MS Detection
Article Snippet: .. Sequencing grade modified trypsin was purchased from Promega Corp. (Madison, WI), ammonium bicarbonate from Aldrich (Milwaukee, WI), and Rhodamine 610 chloride from Exciton (Dayton, OH). .. Protein standards (bovine hemoglobin, fetuin, albumin, alpha casein 1, carbonic anhydrase 2, beta casein, alpha lactalbumin, cytochrome C, ribonuclease A, and horse cytochrome C and myoglobin) and α-cyano-4-hydroxy-cinnamic acid were purchased from Sigma (St. Louis, MO).

Article Title: Profiling constitutive proteolytic events in vivo
Article Snippet: .. The samples were diluted 1:2 with distilled water before digestion overnight by sequencing-grade modified trypsin (Promega) or endoproteinase Glu-C (Roche). .. Further enzymatic activity was inhibited by boiling the samples for 5 min.

Article Title: Proteomics Characterization of Cell Membrane Blebs in Human Retinal Pigment Epithelium Cells *
Article Snippet: .. The gel was stained with GelCode Blue (Pierce); gel slices were excised, destained, reduced with 10 m m DTT for 45 min, alkylated with 55 m m iodoacetamide for 30 min, and digested in situ with sequencing grade modified trypsin (0.1 μg/μl; Promega Biosciences Inc., Madison, WI) following protocols routinely used in our laboratory ( , ); and peptides were subjected to LC MS/MS analysis. .. All LC solvents were purchased from Burdick and Jackson (Muskegon, MI).

Article Title: Cross-Species Analysis of Nicotine-Induced Proteomic Alterations in Pancreatic Cells
Article Snippet: .. Sequencing-grade modified trypsin (V5111) was obtained from Promega (Madison, WI). .. The PaDC cell line, hTERT-HPNE (CRL-4023), was purchased from ATCC (Manassas, VA).

Article Title: Methionine oxidation impairs reverse cholesterol transport by apolipoprotein A-I
Article Snippet: .. HDL was digested with sequencing grade modified trypsin (Promega) or sequencing grade endoproteinase Glu-C (Roche Applied Science) ( , , ). .. MS and MS/MS analyses were performed in the positive ion mode (mass range 200–2,000 Da) with a Thermo-Finnigan LCQ Deca XP Plus instrument ( , , ).

Article Title: The internal interaction in RBBP5 regulates assembly and activity of MLL1 methyltransferase complex
Article Snippet: .. Trypsin digestion was performed with sequencing-grade modified trypsin (Promega, USA) at 37°C overnight. .. The tryptic digested peptides were extracted and desalted with monoSpin C18 columns (GL Science, Japan), followed by separation in a Proxeon EASY-nLC liquid chromatography system (ThermoFisher Scientific, USA) by applying a step-wise gradient of 0–80% acetonitrile in 0.1% formic acid.

Article Title: Top-Down ETD-MS Provides Unreliable Quantitation of Methionine Oxidation
Article Snippet: .. Sequencing-grade modified trypsin was purchased from Promega (Madison, WI, USA). .. Five hundred microliters of 1 mM of protein was incubated with 100 mM H2 O2 , shielded from UV light.

Article Title: KRAS Suppression-Induced Degradation of MYC is Antagonized by a MEK5-ERK5 Compensatory Mechanism
Article Snippet: .. Protein pellets were resuspended in 50 mM HEPES (pH 8.0) and were digested for overnight at 37ºC 24 hr with sequencing grade modified trypsin (Promega). .. Digested peptides were cleaned using PepClean C18 spin columns (Thermo Scientific).

Article Title: The key protein of endosomal mRNP transport Rrm4 binds translational landmark sites of cargo mRNAs
Article Snippet: .. Samples were digested with sequencing‐grade modified trypsin (Promega), and the resulting peptides were separated into fractions by nanoLC (PepMap100 C‐18 RP nanocolumn and UltiMate 3000 liquid chromatography system, Dionex). .. Fractions were spotted with matrix solution on a MALDI plate and subsequently analysed by MALDI‐TOF MS (4800 Proteomics Analyser, AB Sciex).

High Performance Liquid Chromatography:

Article Title: Microfluidic LC Device with Orthogonal Sample Extraction for On-Chip MALDI-MS Detection
Article Snippet: HPLC-grade methanol and acetonitrile were obtained from Fisher Scientific (Fair Lawn, NJ) and deionized water (18 MΩ-cm) was generated in-house using a MilliQ ultrapure water system (Millipore, Bedford, MA). .. Sequencing grade modified trypsin was purchased from Promega Corp. (Madison, WI), ammonium bicarbonate from Aldrich (Milwaukee, WI), and Rhodamine 610 chloride from Exciton (Dayton, OH).

Article Title: A proteomic-based approach to study underlying molecular responses of the small intestine of Wistar rats to genetically modified corn (MON810)
Article Snippet: Liquid chromatography–mass spectrometry (LC–MS/MS) and database search Selected protein spots were excised manually from the gel, and digested with sequencing grade-modified trypsin (Promega, USA) as described by Shevchenko et al. ( ). .. Solubilised peptides were fractionated on a nanoflow HPLC system (Thermo RSLCnano) before online analysis by electrospray ionisation (ESI) mass spectrometry on an amaZon speed ion ETD trap (Bruker Daltonics).

Flow Cytometry:

Article Title: A proteomic-based approach to study underlying molecular responses of the small intestine of Wistar rats to genetically modified corn (MON810)
Article Snippet: Liquid chromatography–mass spectrometry (LC–MS/MS) and database search Selected protein spots were excised manually from the gel, and digested with sequencing grade-modified trypsin (Promega, USA) as described by Shevchenko et al. ( ). .. Peptide separation was performed on a Pepmap C18 reversed phase column (LC Packings), using a 5–85% v/v acetonitrile gradient (in 0.5% v/v formic acid) run over 45 min at a flow rate of 0.2 µl/min.

Chromatography:

Article Title: A proteomic-based approach to study underlying molecular responses of the small intestine of Wistar rats to genetically modified corn (MON810)
Article Snippet: .. Liquid chromatography–mass spectrometry (LC–MS/MS) and database search Selected protein spots were excised manually from the gel, and digested with sequencing grade-modified trypsin (Promega, USA) as described by Shevchenko et al. ( ). .. An ion-trap mass spectrometer (amaZon ETD, Bruker Daltonics, Bremen, Germany) equipped with an on-line nanospray source was used for mass spectrometry data acquisition.

Article Title: Methionine oxidation impairs reverse cholesterol transport by apolipoprotein A-I
Article Snippet: Paragraph title: Liquid Chromatography-Electrospray Ionization-MS. ... HDL was digested with sequencing grade modified trypsin (Promega) or sequencing grade endoproteinase Glu-C (Roche Applied Science) ( , , ).

Buffer Exchange:

Article Title: Profiling constitutive proteolytic events in vivo
Article Snippet: Eventually, unreacted biotin reagent was quenched by the addition of 50 mM glycine for 30 min and subsequently excluded by buffer exchange into 4 M urea, 100 mM Hepes, pH 7.8, and 100 mM NaCl. .. The samples were diluted 1:2 with distilled water before digestion overnight by sequencing-grade modified trypsin (Promega) or endoproteinase Glu-C (Roche).

Transferring:

Article Title: Microfluidic LC Device with Orthogonal Sample Extraction for On-Chip MALDI-MS Detection
Article Snippet: Sequencing grade modified trypsin was purchased from Promega Corp. (Madison, WI), ammonium bicarbonate from Aldrich (Milwaukee, WI), and Rhodamine 610 chloride from Exciton (Dayton, OH). .. The cell Lytic™ NuCLEAR™ extraction kit was from Sigma, and the SPEC-PTC18 and SPEC-PTSCX solid-phase extraction pipette tips were from Agilent.

Liquid Chromatography:

Article Title: The internal interaction in RBBP5 regulates assembly and activity of MLL1 methyltransferase complex
Article Snippet: Trypsin digestion was performed with sequencing-grade modified trypsin (Promega, USA) at 37°C overnight. .. The tryptic digested peptides were extracted and desalted with monoSpin C18 columns (GL Science, Japan), followed by separation in a Proxeon EASY-nLC liquid chromatography system (ThermoFisher Scientific, USA) by applying a step-wise gradient of 0–80% acetonitrile in 0.1% formic acid.

Article Title: Top-Down ETD-MS Provides Unreliable Quantitation of Methionine Oxidation
Article Snippet: Sequencing-grade modified trypsin was purchased from Promega (Madison, WI, USA). .. Sequencing-grade modified trypsin was purchased from Promega (Madison, WI, USA).

Article Title: The key protein of endosomal mRNP transport Rrm4 binds translational landmark sites of cargo mRNAs
Article Snippet: .. Samples were digested with sequencing‐grade modified trypsin (Promega), and the resulting peptides were separated into fractions by nanoLC (PepMap100 C‐18 RP nanocolumn and UltiMate 3000 liquid chromatography system, Dionex). .. Fractions were spotted with matrix solution on a MALDI plate and subsequently analysed by MALDI‐TOF MS (4800 Proteomics Analyser, AB Sciex).

Generated:

Article Title: Microfluidic LC Device with Orthogonal Sample Extraction for On-Chip MALDI-MS Detection
Article Snippet: HPLC-grade methanol and acetonitrile were obtained from Fisher Scientific (Fair Lawn, NJ) and deionized water (18 MΩ-cm) was generated in-house using a MilliQ ultrapure water system (Millipore, Bedford, MA). .. Sequencing grade modified trypsin was purchased from Promega Corp. (Madison, WI), ammonium bicarbonate from Aldrich (Milwaukee, WI), and Rhodamine 610 chloride from Exciton (Dayton, OH).

Tandem Mass Spectroscopy:

Article Title: Proteomics Characterization of Cell Membrane Blebs in Human Retinal Pigment Epithelium Cells *
Article Snippet: .. The gel was stained with GelCode Blue (Pierce); gel slices were excised, destained, reduced with 10 m m DTT for 45 min, alkylated with 55 m m iodoacetamide for 30 min, and digested in situ with sequencing grade modified trypsin (0.1 μg/μl; Promega Biosciences Inc., Madison, WI) following protocols routinely used in our laboratory ( , ); and peptides were subjected to LC MS/MS analysis. .. All LC solvents were purchased from Burdick and Jackson (Muskegon, MI).

Article Title: Methionine oxidation impairs reverse cholesterol transport by apolipoprotein A-I
Article Snippet: HDL was digested with sequencing grade modified trypsin (Promega) or sequencing grade endoproteinase Glu-C (Roche Applied Science) ( , , ). .. MS and MS/MS analyses were performed in the positive ion mode (mass range 200–2,000 Da) with a Thermo-Finnigan LCQ Deca XP Plus instrument ( , , ).

Article Title: The internal interaction in RBBP5 regulates assembly and activity of MLL1 methyltransferase complex
Article Snippet: Trypsin digestion was performed with sequencing-grade modified trypsin (Promega, USA) at 37°C overnight. .. Tandem mass spectrometry (MS/MS) analysis was performed on a Q-Exactive instrument (ThermoFisher Scientific, USA) in a 120-minute gradient.

Sequencing:

Article Title: Microfluidic LC Device with Orthogonal Sample Extraction for On-Chip MALDI-MS Detection
Article Snippet: .. Sequencing grade modified trypsin was purchased from Promega Corp. (Madison, WI), ammonium bicarbonate from Aldrich (Milwaukee, WI), and Rhodamine 610 chloride from Exciton (Dayton, OH). .. Protein standards (bovine hemoglobin, fetuin, albumin, alpha casein 1, carbonic anhydrase 2, beta casein, alpha lactalbumin, cytochrome C, ribonuclease A, and horse cytochrome C and myoglobin) and α-cyano-4-hydroxy-cinnamic acid were purchased from Sigma (St. Louis, MO).

Article Title: Profiling constitutive proteolytic events in vivo
Article Snippet: .. The samples were diluted 1:2 with distilled water before digestion overnight by sequencing-grade modified trypsin (Promega) or endoproteinase Glu-C (Roche). .. Further enzymatic activity was inhibited by boiling the samples for 5 min.

Article Title: Proteomics Characterization of Cell Membrane Blebs in Human Retinal Pigment Epithelium Cells *
Article Snippet: .. The gel was stained with GelCode Blue (Pierce); gel slices were excised, destained, reduced with 10 m m DTT for 45 min, alkylated with 55 m m iodoacetamide for 30 min, and digested in situ with sequencing grade modified trypsin (0.1 μg/μl; Promega Biosciences Inc., Madison, WI) following protocols routinely used in our laboratory ( , ); and peptides were subjected to LC MS/MS analysis. .. All LC solvents were purchased from Burdick and Jackson (Muskegon, MI).

Article Title: A proteomic-based approach to study underlying molecular responses of the small intestine of Wistar rats to genetically modified corn (MON810)
Article Snippet: .. Liquid chromatography–mass spectrometry (LC–MS/MS) and database search Selected protein spots were excised manually from the gel, and digested with sequencing grade-modified trypsin (Promega, USA) as described by Shevchenko et al. ( ). .. An ion-trap mass spectrometer (amaZon ETD, Bruker Daltonics, Bremen, Germany) equipped with an on-line nanospray source was used for mass spectrometry data acquisition.

Article Title: Cross-Species Analysis of Nicotine-Induced Proteomic Alterations in Pancreatic Cells
Article Snippet: .. Sequencing-grade modified trypsin (V5111) was obtained from Promega (Madison, WI). .. The PaDC cell line, hTERT-HPNE (CRL-4023), was purchased from ATCC (Manassas, VA).

Article Title: Methionine oxidation impairs reverse cholesterol transport by apolipoprotein A-I
Article Snippet: .. HDL was digested with sequencing grade modified trypsin (Promega) or sequencing grade endoproteinase Glu-C (Roche Applied Science) ( , , ). .. MS and MS/MS analyses were performed in the positive ion mode (mass range 200–2,000 Da) with a Thermo-Finnigan LCQ Deca XP Plus instrument ( , , ).

Article Title: The internal interaction in RBBP5 regulates assembly and activity of MLL1 methyltransferase complex
Article Snippet: .. Trypsin digestion was performed with sequencing-grade modified trypsin (Promega, USA) at 37°C overnight. .. The tryptic digested peptides were extracted and desalted with monoSpin C18 columns (GL Science, Japan), followed by separation in a Proxeon EASY-nLC liquid chromatography system (ThermoFisher Scientific, USA) by applying a step-wise gradient of 0–80% acetonitrile in 0.1% formic acid.

Article Title: Top-Down ETD-MS Provides Unreliable Quantitation of Methionine Oxidation
Article Snippet: .. Sequencing-grade modified trypsin was purchased from Promega (Madison, WI, USA). .. Five hundred microliters of 1 mM of protein was incubated with 100 mM H2 O2 , shielded from UV light.

Article Title: KRAS Suppression-Induced Degradation of MYC is Antagonized by a MEK5-ERK5 Compensatory Mechanism
Article Snippet: .. Protein pellets were resuspended in 50 mM HEPES (pH 8.0) and were digested for overnight at 37ºC 24 hr with sequencing grade modified trypsin (Promega). .. Digested peptides were cleaned using PepClean C18 spin columns (Thermo Scientific).

Article Title: The key protein of endosomal mRNP transport Rrm4 binds translational landmark sites of cargo mRNAs
Article Snippet: .. Samples were digested with sequencing‐grade modified trypsin (Promega), and the resulting peptides were separated into fractions by nanoLC (PepMap100 C‐18 RP nanocolumn and UltiMate 3000 liquid chromatography system, Dionex). .. Fractions were spotted with matrix solution on a MALDI plate and subsequently analysed by MALDI‐TOF MS (4800 Proteomics Analyser, AB Sciex).

Affinity Purification:

Article Title: The key protein of endosomal mRNP transport Rrm4 binds translational landmark sites of cargo mRNAs
Article Snippet: Paragraph title: Preliminary affinity purification experiments using Rrm4‐GfpTT as bait ... Samples were digested with sequencing‐grade modified trypsin (Promega), and the resulting peptides were separated into fractions by nanoLC (PepMap100 C‐18 RP nanocolumn and UltiMate 3000 liquid chromatography system, Dionex).

Isolation:

Article Title: Sensitive Determination of Proteolytic Proteoforms in Limited Microscale Proteome Samples *
Article Snippet: Subsequently, trypsin (1 mg/ml, Promega, cat. no. V5113) was introduced and incubated at 37 °C overnight. .. Finally, the tagged-internal peptides were isolated by stage-tips.

Protein Concentration:

Article Title: Binding of calcium ions and SNAP-25 to the hexa EF-hand protein secretagogin
Article Snippet: Secretagogin was dissolved in 50 mM Tris/HCl and 150 mM KCl, containing either 1 mM CaCl2 or 1 mM EDTA at a protein concentration of 1 mg/ml. .. Sequencing grade modified trypsin (Promega) was dissolved in 50 mM ammonium bicarbonate, yielding a stock solution with a concentration of 0.5 mg/ml.

Labeling:

Article Title: Sensitive Determination of Proteolytic Proteoforms in Limited Microscale Proteome Samples *
Article Snippet: Dimethyl labeling was used to block amine groups within the proteome. .. Subsequently, trypsin (1 mg/ml, Promega, cat. no. V5113) was introduced and incubated at 37 °C overnight.

Purification:

Article Title: The key protein of endosomal mRNP transport Rrm4 binds translational landmark sites of cargo mRNAs
Article Snippet: Preliminary purification experiments were performed using a strain expressing Rrm4 fused C‐terminally to Gfp and tandem affinity purification (TAP) tag . .. Samples were digested with sequencing‐grade modified trypsin (Promega), and the resulting peptides were separated into fractions by nanoLC (PepMap100 C‐18 RP nanocolumn and UltiMate 3000 liquid chromatography system, Dionex).

SDS Page:

Article Title: Proteomics Characterization of Cell Membrane Blebs in Human Retinal Pigment Epithelium Cells *
Article Snippet: Proteins (10 μg) were separated by 4–20% gradient SDS-PAGE (Invitrogen). .. The gel was stained with GelCode Blue (Pierce); gel slices were excised, destained, reduced with 10 m m DTT for 45 min, alkylated with 55 m m iodoacetamide for 30 min, and digested in situ with sequencing grade modified trypsin (0.1 μg/μl; Promega Biosciences Inc., Madison, WI) following protocols routinely used in our laboratory ( , ); and peptides were subjected to LC MS/MS analysis.

Article Title: The internal interaction in RBBP5 regulates assembly and activity of MLL1 methyltransferase complex
Article Snippet: The reaction products were separated by SDS-PAGE on 8–10% Tris-glycine gels and visualized by staining with Coomassie blue. .. Trypsin digestion was performed with sequencing-grade modified trypsin (Promega, USA) at 37°C overnight.

Article Title: The key protein of endosomal mRNP transport Rrm4 binds translational landmark sites of cargo mRNAs
Article Snippet: We analysed the eluted fractions by mass spectrometry either as complex mixture in solution (Fig C) or by cutting individual bands from an SDS–PAGE (Fig D). .. Samples were digested with sequencing‐grade modified trypsin (Promega), and the resulting peptides were separated into fractions by nanoLC (PepMap100 C‐18 RP nanocolumn and UltiMate 3000 liquid chromatography system, Dionex).

Software:

Article Title: The key protein of endosomal mRNP transport Rrm4 binds translational landmark sites of cargo mRNAs
Article Snippet: Samples were digested with sequencing‐grade modified trypsin (Promega), and the resulting peptides were separated into fractions by nanoLC (PepMap100 C‐18 RP nanocolumn and UltiMate 3000 liquid chromatography system, Dionex). .. Mass spectrometry data were searched against the MUMDB (Munich Ustilago Maydis Data Base; 6,785 entries; IBIS Institute of Bioinformatics and Systems, German Research Center for Environmental Health) using Masc ot embedded into GPS Explorer software (AB Sciex).

Selection:

Article Title: Profiling constitutive proteolytic events in vivo
Article Snippet: 1 h. This NHS-reactive reagent is specific for the N-terminal of the proteins, i.e. native N-termini and proteolytic cleavage sites, owing to the previous blocking of cysteine and lysine side chains, and the biotin tag of the molecule allows for positive selection by immobilized streptavidin. .. The samples were diluted 1:2 with distilled water before digestion overnight by sequencing-grade modified trypsin (Promega) or endoproteinase Glu-C (Roche).

In Situ:

Article Title: Proteomics Characterization of Cell Membrane Blebs in Human Retinal Pigment Epithelium Cells *
Article Snippet: .. The gel was stained with GelCode Blue (Pierce); gel slices were excised, destained, reduced with 10 m m DTT for 45 min, alkylated with 55 m m iodoacetamide for 30 min, and digested in situ with sequencing grade modified trypsin (0.1 μg/μl; Promega Biosciences Inc., Madison, WI) following protocols routinely used in our laboratory ( , ); and peptides were subjected to LC MS/MS analysis. .. All LC solvents were purchased from Burdick and Jackson (Muskegon, MI).

Concentration Assay:

Article Title: The internal interaction in RBBP5 regulates assembly and activity of MLL1 methyltransferase complex
Article Snippet: Then Sulfo-NHS (N -hydroxysulfosuccinimide) was added to the reaction mixture at a final concentration of 2 mM, followed by incubation at room temperature for 2 h. Finally, the reaction was quenched by adding 20 mM Tris (pH 8.0). .. Trypsin digestion was performed with sequencing-grade modified trypsin (Promega, USA) at 37°C overnight.

Article Title: Binding of calcium ions and SNAP-25 to the hexa EF-hand protein secretagogin
Article Snippet: .. Sequencing grade modified trypsin (Promega) was dissolved in 50 mM ammonium bicarbonate, yielding a stock solution with a concentration of 0.5 mg/ml. ..

Article Title: Sensitive Determination of Proteolytic Proteoforms in Limited Microscale Proteome Samples *
Article Snippet: Briefly, HeLa cells were first lysed with 6 m guanidine hydrochloride (GuHCl, Fisher BioReagents, cat. no. BP178–1) to a final concentration of 3 m in a 1.5 ml protein Lobind tube (Eppendorf, cat. no. 022431081). .. Subsequently, trypsin (1 mg/ml, Promega, cat. no. V5113) was introduced and incubated at 37 °C overnight.

Fractionation:

Article Title: Sensitive Determination of Proteolytic Proteoforms in Limited Microscale Proteome Samples *
Article Snippet: Paragraph title: High-pH Reversed Phase Fractionation ... Subsequently, trypsin (1 mg/ml, Promega, cat. no. V5113) was introduced and incubated at 37 °C overnight.

Staining:

Article Title: Proteomics Characterization of Cell Membrane Blebs in Human Retinal Pigment Epithelium Cells *
Article Snippet: .. The gel was stained with GelCode Blue (Pierce); gel slices were excised, destained, reduced with 10 m m DTT for 45 min, alkylated with 55 m m iodoacetamide for 30 min, and digested in situ with sequencing grade modified trypsin (0.1 μg/μl; Promega Biosciences Inc., Madison, WI) following protocols routinely used in our laboratory ( , ); and peptides were subjected to LC MS/MS analysis. .. All LC solvents were purchased from Burdick and Jackson (Muskegon, MI).

Article Title: Cross-Species Analysis of Nicotine-Induced Proteomic Alterations in Pancreatic Cells
Article Snippet: SeeBluePlus2 Pre-Stained standard (LC5925), LDS (lithium dodecyl sulfate) sample buffer (NP0008), NuPAGE 4-12% Bis-Tris polyacrylamide gels (NP0335), SimplyBlue Coomassie stain (LC0665), and MES-SDS (2-(N-morpholino) ethanesulfonic acid-sodium dodecyl sulfate) electrophoresis buffer (NP002) were from Invitrogen (Carlsbad, CA). (-)-Nicotine (≥99%) (N3876) was purchased from Sigma (St. Louis, MO). .. Sequencing-grade modified trypsin (V5111) was obtained from Promega (Madison, WI).

Article Title: The internal interaction in RBBP5 regulates assembly and activity of MLL1 methyltransferase complex
Article Snippet: The reaction products were separated by SDS-PAGE on 8–10% Tris-glycine gels and visualized by staining with Coomassie blue. .. Trypsin digestion was performed with sequencing-grade modified trypsin (Promega, USA) at 37°C overnight.

Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94
    Promega two step digestion approach
    Two Step Digestion Approach, supplied by Promega, used in various techniques. Bioz Stars score: 94/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/two step digestion approach/product/Promega
    Average 94 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    two step digestion approach - by Bioz Stars, 2020-04
    94/100 stars
      Buy from Supplier

    99
    Promega sequencing grade modified trypsin
    Sequencing Grade Modified Trypsin, supplied by Promega, used in various techniques. Bioz Stars score: 99/100, based on 43 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sequencing grade modified trypsin/product/Promega
    Average 99 stars, based on 43 article reviews
    Price from $9.99 to $1999.99
    sequencing grade modified trypsin - by Bioz Stars, 2020-04
    99/100 stars
      Buy from Supplier

    Image Search Results