dispase  (Worthington Biochemical)


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    Name:
    Neutral Protease Dispase Purified
    Description:
    Animal Origin Free Chromatographically purified A lyophilized powder
    Catalog Number:
    LS02100
    Price:
    70
    Source:
    Bacillus polymyxa
    Size:
    10 mg
    Cas Number:
    42613.33.2
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    Structured Review

    Worthington Biochemical dispase
    <t>Dispase</t> treatment reduces capsid redistribution to the nucleus. (A) pH-dependent dispase cleavage of AdV5 capsid hexon. Purified AdV5 capsids were treated with dispase at pH 7.4 or pH 4.4 or kept at pH 4.4 alone before being boiled in PSB and subjected
    Animal Origin Free Chromatographically purified A lyophilized powder
    https://www.bioz.com/result/dispase/product/Worthington Biochemical
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    dispase - by Bioz Stars, 2021-10
    97/100 stars

    Images

    1) Product Images from "Conformational Changes in the Adenovirus Hexon Subunit Responsible for Regulating Cytoplasmic Dynein Recruitment"

    Article Title: Conformational Changes in the Adenovirus Hexon Subunit Responsible for Regulating Cytoplasmic Dynein Recruitment

    Journal: Journal of Virology

    doi: 10.1128/JVI.02889-14

    Dispase treatment reduces capsid redistribution to the nucleus. (A) pH-dependent dispase cleavage of AdV5 capsid hexon. Purified AdV5 capsids were treated with dispase at pH 7.4 or pH 4.4 or kept at pH 4.4 alone before being boiled in PSB and subjected
    Figure Legend Snippet: Dispase treatment reduces capsid redistribution to the nucleus. (A) pH-dependent dispase cleavage of AdV5 capsid hexon. Purified AdV5 capsids were treated with dispase at pH 7.4 or pH 4.4 or kept at pH 4.4 alone before being boiled in PSB and subjected

    Techniques Used: Purification

    Dispase hydrolysis inhibits low-pH effects on hexon structure and dynein binding. (A) Purified hexon was tested for dissociation after dispase treatment. Dispase exposure of hexon at low pH resulted in two major digestion products (85 kDa [I] and 15 kDa
    Figure Legend Snippet: Dispase hydrolysis inhibits low-pH effects on hexon structure and dynein binding. (A) Purified hexon was tested for dissociation after dispase treatment. Dispase exposure of hexon at low pH resulted in two major digestion products (85 kDa [I] and 15 kDa

    Techniques Used: Binding Assay, Purification

    The AdV5 hexon HVR1 mutant is inhibited in dynein binding and cytoplasmic transport. (A) Dispase insensitivity of mutant hexon. Immunopurified hexons of wild-type (WT) AdV5, AdV5HVR48(1), and WT AdV48 were tested for dissociation after dispase treatment.
    Figure Legend Snippet: The AdV5 hexon HVR1 mutant is inhibited in dynein binding and cytoplasmic transport. (A) Dispase insensitivity of mutant hexon. Immunopurified hexons of wild-type (WT) AdV5, AdV5HVR48(1), and WT AdV48 were tested for dissociation after dispase treatment.

    Techniques Used: Mutagenesis, Binding Assay

    2) Product Images from "Isolation of mitochondrial subpopulations from skeletal muscle: optimizing recovery and preserving integrity."

    Article Title: Isolation of mitochondrial subpopulations from skeletal muscle: optimizing recovery and preserving integrity.

    Journal: Acta physiologica (Oxford, England)

    doi: 10.1111/apha.13182

    Two distinct cell fragments found within a myofibrillar pellet after homogenization and centrifugation of dispase pellet (Scale line = 2μm) in panel (a) and (b). The sarcolemma and interfibrillar (IFM) mitochondria are identified.
    Figure Legend Snippet: Two distinct cell fragments found within a myofibrillar pellet after homogenization and centrifugation of dispase pellet (Scale line = 2μm) in panel (a) and (b). The sarcolemma and interfibrillar (IFM) mitochondria are identified.

    Techniques Used: Homogenization, Centrifugation

    Two distinct fragments of a cell found within the skeletal muscle pellet treated with dispase protease (Scale line = 2μm) in panel (a) and (b). The sarcolemma, subsarcolemmal (SSM) and interfibrillar (IFM) mitochondria are identified.
    Figure Legend Snippet: Two distinct fragments of a cell found within the skeletal muscle pellet treated with dispase protease (Scale line = 2μm) in panel (a) and (b). The sarcolemma, subsarcolemmal (SSM) and interfibrillar (IFM) mitochondria are identified.

    Techniques Used:

    Related Articles

    other:

    Article Title: Coronary blood vessels from distinct origins converge to equivalent states during mouse and human development
    Article Snippet: Hearts were then dissociated in a 600 μl mix consisting of 500 U/ml collagenase IV (Worthington #LS004186), 1.2 U/ml dispase (Worthington #LS02100), 32 U/ml DNase I (Worthington #LS002007), and sterile DPBS with Mg2+ and Ca2+ at 37 degrees for 45 minutes and resuspended by pipetting every 5 minutes.

    Article Title: Human anogenital monocyte-derived dendritic cells and langerin+cDC2 are major HIV target cells
    Article Snippet: The meshed skin was placed in RPMI1640 (Lonza, Switzerland) with 0.14 U/ml dispase (neutral protease, Worthington Industries, Columbus, OH, USA) and 50 μg/mL Gentamicin (Sigma-Aldrich, St Louis, MO, USA) and rotated at 4 °C overnight.

    Article Title: Microglial NF-κB drives tau spreading and toxicity in a mouse model of tauopathy
    Article Snippet: Dissected brains were chilled on ice and minced in digestion media containing with 0.2% collagenase type 3 (LS004182, Worthington) and 3 U/mL dispase (LS02104, Worthington).

    Incubation:

    Article Title: Comparison of quinazoline and benzoylpyrazoline chemotypes targeting the CaVα-β interaction as antagonists of the N-type CaV2.2 channel
    Article Snippet: .. DRG were then collected, trimmed at their roots, and enzymatically digested in 3 mL bicarbonate-free, serum-free, sterile DMEM (Cat# 11,965, Thermo Fisher Scientific, Waltham, MA) solution containing neutral protease (3.125 mg.ml-1, CatLS02104; Worthington, Lakewood, NJ) and collagenase type I (5 mg/mL, Cat# LS004194, Worthington, Lakewood, NJ) and incubated for 60 minutes at 37°C under gentile agitation. ..

    Article Title: Non-SUMOylated CRMP2 decreases NaV1.7 currents via the endocytic proteins Numb, Nedd4-2 and Eps15
    Article Snippet: .. Dorsal root ganglia (DRG) were quickly removed, trimmed at their roots, and enzymatically digested in 3 mL bicarbonate-free, serum-free, sterile DMEM (Cat# 11,965, Thermo Fisher Scientific, Waltham, MA) solution containing neutral protease (1.87 mg/ml, CatLS02104; Worthington, Lakewood, NJ) and collagenase type I (3 mg/mL, Cat# LS004194, Worthington, Lakewood, NJ) and incubated for 50 min at 37 °C under gentle agitation. ..

    Article Title: Gut-licensed IFNγ+ NK cells drive LAMP1+TRAIL+ anti-inflammatory astrocytes
    Article Snippet: .. Tissue was incubated for 30 min at 37 °C in 500 μl of digestion mix composed of 0.5 mg ml−1 of collagenase P (Sigma-Aldrich, 11213865001), 0.5 mg ml−1 of dispase (Worthington, LS02104), and 125 U ml−1 of DNase I (Sigma-Aldrich, 10104159001) in RPMI-1640. ..

    Article Title: Cyclic Stretch of Either PNS or CNS Located Nerves Can Stimulate Neurite Outgrowth
    Article Snippet: .. DRG were spun down at 300× g for 1 min. HBSS was replaced by 500 μL of a digestion mixture of 2.5 mg/mL collagenase (Merck KGaA, C9407, Darmstadt, Germany) and 5 mg/mL neutral protease (dispase) (Worthington Biochemical Corp., LS02104, Lakewood, NJ, USA) in HBSS and incubated at 37 °C for 35 min by gently shaking every 10 min. Then, the DRG were centrifuged at 300× g for 2 min, and the digestion solution was discarded. ..

    Mouse Assay:

    Article Title: 3-hydroxy-L-kynurenamine is an immunomodulatory biogenic amine
    Article Snippet: .. To obtain primary fibroblast reticular cells (FRC) lymph nodes (axillary, inguinal, mesenteric, and brachial) were collected from C57BL/6J mice and digested with 0.8 mg/ml Dispase (Worthington Biochemical corporation, cat# LS02100, 0.2 mg/ml Collagenase P (Roche, cat# 11213857 001) and 0.1 mg/ml DNase I (Millipore Sigma, cat# 260913-10MU) at 37 °C on a shaker for 60 min. At 15-min intervals, digested tissue was pipetted to facility dissociation and dispersed single cells were collected. ..

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  • 97
    Worthington Biochemical dispase
    <t>Dispase</t> treatment reduces capsid redistribution to the nucleus. (A) pH-dependent dispase cleavage of AdV5 capsid hexon. Purified AdV5 capsids were treated with dispase at pH 7.4 or pH 4.4 or kept at pH 4.4 alone before being boiled in PSB and subjected
    Dispase, supplied by Worthington Biochemical, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dispase/product/Worthington Biochemical
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    dispase - by Bioz Stars, 2021-10
    97/100 stars
      Buy from Supplier

    99
    Worthington Biochemical collagenase type 2
    Defective contractile function of S3KO aorta. A through C, PE‐induced contraction of WT and S3KO aortic rings treated with placebo (A), 200 μmol/L l‐NAME (B), or 200 μmol/L aminoguanidine (C) (n=4 per group; duplicate experiments were performed). Contraction was measured at PE concentrations ranging from 10 −10 to 10 −3 mol/L, and concentration–response curves were constructed. Note that contraction is expressed as the percentage of maximal contraction induced by incubation of aortic rings in 80 mmol/L high potassium (Hi‐K + ) buffer. For placebo and l‐NAME treatment, endothelium‐intact aortic segments were used, whereas endothelium‐denuded aortas were used in aminoguanidine treatment to eliminate the vasorelaxant effect of eNOS‐derived NO. D, pEC 50 of WT and S3KO aortas in response to PE, indicating tissue sensitivity toward the drug. Note that pEC 50 is the negative logarithm of EC 50 , which refers to the concentration of a drug that induces a half‐maximal response. E, Maximal contraction force generated when incubated with 80 mmol/L Hi‐K + buffer. F, Aortic mRNA level of AngII type 1a (AT1a), type 1b (AT1b), and <t>type</t> 2 (AT2) receptors in WT and S3KO mice (n=3 per group; triplicate experiments were performed). Gene expression was normalized to RPL27. G through I, AngII‐induced contraction in ex vivo aortic segments treated with placebo (G), 200 μmol/L l‐NAME (H), or 200 μmol/L aminoguanidine (I) (n=5 per group; duplicate experiments were performed). Note that AngII elicited a rapid and transient contraction of infrarenal aortic segments at a concentration of 1 μg/mL, whereas minimal (
    Collagenase Type 2, supplied by Worthington Biochemical, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/collagenase type 2/product/Worthington Biochemical
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    95
    Worthington Biochemical dnase i
    Purification procedure. (a ) Strategy. Cartoon (top left): chromatin loop with nucleosomes (green circles) tethered to a polymerizing complex (oval) attached to the substructure (brown). Cells are permeabilized, in some cases a run-on performed in [ 32 P]UTP so nascent RNA can be tracked, nuclei are washed with NP40, most chromatin detached with a nuclease (here, DNase I), chromatin-depleted nuclei resuspended in NLB, and polymerizing complexes released from the substructure with caspases. After pelleting, chromatin associated with polymerizing complexes in the supernatant is degraded with DNase I, and complexes partially resolved in 2D gels (using “blue native” and “native” gels in the first and second dimensions); rough positions of complexes (and a control region, c) are shown. Finally, different regions are excised, and their content analyzed by mass spectrometry. ( b ) Recovery of [ 32 P]RNA, after including a “run-on”. Fractions correspond to those at the same level in ( a ). ( c)  “Run-on” activity assayed later during fractionation (as in  a , but without run-on at beginning). Different fractions, with names as in ( a ), were allowed to extend transcripts by
    Dnase I, supplied by Worthington Biochemical, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Worthington Biochemical rnase t1
    LC-MS analysis of RNaseT1 digests of yeast tRNA Phe−1 . ( a ) Base peak chromatogram of the <t>RNase</t> T1 digest of the yeast tRNA Phe−1 preparation (200 fmol). Chromatography was performed as described in the ‘Materials and Methods’ section. Sixteen major oligoribonucleotide peaks, indicated by arrows with peak numbers, are assigned to the fragments of yeast tRNA Phe−1 , tRNA Phe−2 , tRNA Lys−2 , or tRNA Tyr (see Table 1 ). ( b and c ) A typical MS spectrum of the RNase T1 digest of tRNA Phe−1 ; (b) [AUUUAm 2 G > p] 2− and (c) [ACmUGmAAyWAΨUm 5 CUG > p] 3− .
    Rnase T1, supplied by Worthington Biochemical, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Dispase treatment reduces capsid redistribution to the nucleus. (A) pH-dependent dispase cleavage of AdV5 capsid hexon. Purified AdV5 capsids were treated with dispase at pH 7.4 or pH 4.4 or kept at pH 4.4 alone before being boiled in PSB and subjected

    Journal: Journal of Virology

    Article Title: Conformational Changes in the Adenovirus Hexon Subunit Responsible for Regulating Cytoplasmic Dynein Recruitment

    doi: 10.1128/JVI.02889-14

    Figure Lengend Snippet: Dispase treatment reduces capsid redistribution to the nucleus. (A) pH-dependent dispase cleavage of AdV5 capsid hexon. Purified AdV5 capsids were treated with dispase at pH 7.4 or pH 4.4 or kept at pH 4.4 alone before being boiled in PSB and subjected

    Article Snippet: For controlled proteolysis, hexon or AdV capsids were incubated with a 6-fold molar excess of dispase (neutral protease; Worthington Biochemicals, Freehold, NJ) over hexon in dispase buffer (100 mM Trizma-maleate, 120 mM NaCl, 2.5 mM CaCl2 ), and the pH was adjusted to either pH 7.4 (neutral) or pH 4.4 (acidic) for 5 min at 37°C until the reaction was stopped by the addition of either protein sample buffer (PSB) or 5 mM EDTA to the mixture.

    Techniques: Purification

    Dispase hydrolysis inhibits low-pH effects on hexon structure and dynein binding. (A) Purified hexon was tested for dissociation after dispase treatment. Dispase exposure of hexon at low pH resulted in two major digestion products (85 kDa [I] and 15 kDa

    Journal: Journal of Virology

    Article Title: Conformational Changes in the Adenovirus Hexon Subunit Responsible for Regulating Cytoplasmic Dynein Recruitment

    doi: 10.1128/JVI.02889-14

    Figure Lengend Snippet: Dispase hydrolysis inhibits low-pH effects on hexon structure and dynein binding. (A) Purified hexon was tested for dissociation after dispase treatment. Dispase exposure of hexon at low pH resulted in two major digestion products (85 kDa [I] and 15 kDa

    Article Snippet: For controlled proteolysis, hexon or AdV capsids were incubated with a 6-fold molar excess of dispase (neutral protease; Worthington Biochemicals, Freehold, NJ) over hexon in dispase buffer (100 mM Trizma-maleate, 120 mM NaCl, 2.5 mM CaCl2 ), and the pH was adjusted to either pH 7.4 (neutral) or pH 4.4 (acidic) for 5 min at 37°C until the reaction was stopped by the addition of either protein sample buffer (PSB) or 5 mM EDTA to the mixture.

    Techniques: Binding Assay, Purification

    The AdV5 hexon HVR1 mutant is inhibited in dynein binding and cytoplasmic transport. (A) Dispase insensitivity of mutant hexon. Immunopurified hexons of wild-type (WT) AdV5, AdV5HVR48(1), and WT AdV48 were tested for dissociation after dispase treatment.

    Journal: Journal of Virology

    Article Title: Conformational Changes in the Adenovirus Hexon Subunit Responsible for Regulating Cytoplasmic Dynein Recruitment

    doi: 10.1128/JVI.02889-14

    Figure Lengend Snippet: The AdV5 hexon HVR1 mutant is inhibited in dynein binding and cytoplasmic transport. (A) Dispase insensitivity of mutant hexon. Immunopurified hexons of wild-type (WT) AdV5, AdV5HVR48(1), and WT AdV48 were tested for dissociation after dispase treatment.

    Article Snippet: For controlled proteolysis, hexon or AdV capsids were incubated with a 6-fold molar excess of dispase (neutral protease; Worthington Biochemicals, Freehold, NJ) over hexon in dispase buffer (100 mM Trizma-maleate, 120 mM NaCl, 2.5 mM CaCl2 ), and the pH was adjusted to either pH 7.4 (neutral) or pH 4.4 (acidic) for 5 min at 37°C until the reaction was stopped by the addition of either protein sample buffer (PSB) or 5 mM EDTA to the mixture.

    Techniques: Mutagenesis, Binding Assay

    Defective contractile function of S3KO aorta. A through C, PE‐induced contraction of WT and S3KO aortic rings treated with placebo (A), 200 μmol/L l‐NAME (B), or 200 μmol/L aminoguanidine (C) (n=4 per group; duplicate experiments were performed). Contraction was measured at PE concentrations ranging from 10 −10 to 10 −3 mol/L, and concentration–response curves were constructed. Note that contraction is expressed as the percentage of maximal contraction induced by incubation of aortic rings in 80 mmol/L high potassium (Hi‐K + ) buffer. For placebo and l‐NAME treatment, endothelium‐intact aortic segments were used, whereas endothelium‐denuded aortas were used in aminoguanidine treatment to eliminate the vasorelaxant effect of eNOS‐derived NO. D, pEC 50 of WT and S3KO aortas in response to PE, indicating tissue sensitivity toward the drug. Note that pEC 50 is the negative logarithm of EC 50 , which refers to the concentration of a drug that induces a half‐maximal response. E, Maximal contraction force generated when incubated with 80 mmol/L Hi‐K + buffer. F, Aortic mRNA level of AngII type 1a (AT1a), type 1b (AT1b), and type 2 (AT2) receptors in WT and S3KO mice (n=3 per group; triplicate experiments were performed). Gene expression was normalized to RPL27. G through I, AngII‐induced contraction in ex vivo aortic segments treated with placebo (G), 200 μmol/L l‐NAME (H), or 200 μmol/L aminoguanidine (I) (n=5 per group; duplicate experiments were performed). Note that AngII elicited a rapid and transient contraction of infrarenal aortic segments at a concentration of 1 μg/mL, whereas minimal (

    Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

    Article Title: SMAD3 Deficiency Promotes Inflammatory Aortic Aneurysms in Angiotensin II-Infused Mice Via Activation of iNOS

    doi: 10.1161/JAHA.113.000269

    Figure Lengend Snippet: Defective contractile function of S3KO aorta. A through C, PE‐induced contraction of WT and S3KO aortic rings treated with placebo (A), 200 μmol/L l‐NAME (B), or 200 μmol/L aminoguanidine (C) (n=4 per group; duplicate experiments were performed). Contraction was measured at PE concentrations ranging from 10 −10 to 10 −3 mol/L, and concentration–response curves were constructed. Note that contraction is expressed as the percentage of maximal contraction induced by incubation of aortic rings in 80 mmol/L high potassium (Hi‐K + ) buffer. For placebo and l‐NAME treatment, endothelium‐intact aortic segments were used, whereas endothelium‐denuded aortas were used in aminoguanidine treatment to eliminate the vasorelaxant effect of eNOS‐derived NO. D, pEC 50 of WT and S3KO aortas in response to PE, indicating tissue sensitivity toward the drug. Note that pEC 50 is the negative logarithm of EC 50 , which refers to the concentration of a drug that induces a half‐maximal response. E, Maximal contraction force generated when incubated with 80 mmol/L Hi‐K + buffer. F, Aortic mRNA level of AngII type 1a (AT1a), type 1b (AT1b), and type 2 (AT2) receptors in WT and S3KO mice (n=3 per group; triplicate experiments were performed). Gene expression was normalized to RPL27. G through I, AngII‐induced contraction in ex vivo aortic segments treated with placebo (G), 200 μmol/L l‐NAME (H), or 200 μmol/L aminoguanidine (I) (n=5 per group; duplicate experiments were performed). Note that AngII elicited a rapid and transient contraction of infrarenal aortic segments at a concentration of 1 μg/mL, whereas minimal (

    Article Snippet: Briefly, the whole aorta was dissected out from its origin at the left ventricle to the iliac bifurcation, denuded of periaortic fat, cut into small pieces (≈2‐mm segments), and incubated in 5 mL of Dulbecco's modified Eagle's medium (DMEM) containing 10% fetal bovine serum (#16000036; Life Technologies) and 1.36 mg/mL of type II collagenase (#LS004174; Worthington Biochemical Corporation) in agitation at 115 rpm for 2 hours at 37°C.

    Techniques: Concentration Assay, Construct, Incubation, Derivative Assay, Generated, Mouse Assay, Expressing, Ex Vivo

    Purification procedure. (a ) Strategy. Cartoon (top left): chromatin loop with nucleosomes (green circles) tethered to a polymerizing complex (oval) attached to the substructure (brown). Cells are permeabilized, in some cases a run-on performed in [ 32 P]UTP so nascent RNA can be tracked, nuclei are washed with NP40, most chromatin detached with a nuclease (here, DNase I), chromatin-depleted nuclei resuspended in NLB, and polymerizing complexes released from the substructure with caspases. After pelleting, chromatin associated with polymerizing complexes in the supernatant is degraded with DNase I, and complexes partially resolved in 2D gels (using “blue native” and “native” gels in the first and second dimensions); rough positions of complexes (and a control region, c) are shown. Finally, different regions are excised, and their content analyzed by mass spectrometry. ( b ) Recovery of [ 32 P]RNA, after including a “run-on”. Fractions correspond to those at the same level in ( a ). ( c)  “Run-on” activity assayed later during fractionation (as in  a , but without run-on at beginning). Different fractions, with names as in ( a ), were allowed to extend transcripts by

    Journal: Nature methods

    Article Title: The proteomes of transcription factories containing RNA polymerases I, II or III

    doi: 10.1038/nmeth.1705

    Figure Lengend Snippet: Purification procedure. (a ) Strategy. Cartoon (top left): chromatin loop with nucleosomes (green circles) tethered to a polymerizing complex (oval) attached to the substructure (brown). Cells are permeabilized, in some cases a run-on performed in [ 32 P]UTP so nascent RNA can be tracked, nuclei are washed with NP40, most chromatin detached with a nuclease (here, DNase I), chromatin-depleted nuclei resuspended in NLB, and polymerizing complexes released from the substructure with caspases. After pelleting, chromatin associated with polymerizing complexes in the supernatant is degraded with DNase I, and complexes partially resolved in 2D gels (using “blue native” and “native” gels in the first and second dimensions); rough positions of complexes (and a control region, c) are shown. Finally, different regions are excised, and their content analyzed by mass spectrometry. ( b ) Recovery of [ 32 P]RNA, after including a “run-on”. Fractions correspond to those at the same level in ( a ). ( c) “Run-on” activity assayed later during fractionation (as in a , but without run-on at beginning). Different fractions, with names as in ( a ), were allowed to extend transcripts by

    Article Snippet: Resuspended nuclei were digested (30 min; 33°C) with either (i) DNase I (protease- and RNase-free; Worthington;10 units/107 cells in 100 μl PB plus 0.5 mM CaCl2 ), or (ii) Hae III (Invitrogen; 1000 units/107 cells), or (iii) Hind III (New England Biolabs; 1000 units/107 cells) in PB; reactions were stopped by adding EDTA to 2.5 mM and cooling in iced water.

    Techniques: Purification, Mass Spectrometry, Activity Assay, Fractionation

    LC-MS analysis of RNaseT1 digests of yeast tRNA Phe−1 . ( a ) Base peak chromatogram of the RNase T1 digest of the yeast tRNA Phe−1 preparation (200 fmol). Chromatography was performed as described in the ‘Materials and Methods’ section. Sixteen major oligoribonucleotide peaks, indicated by arrows with peak numbers, are assigned to the fragments of yeast tRNA Phe−1 , tRNA Phe−2 , tRNA Lys−2 , or tRNA Tyr (see Table 1 ). ( b and c ) A typical MS spectrum of the RNase T1 digest of tRNA Phe−1 ; (b) [AUUUAm 2 G > p] 2− and (c) [ACmUGmAAyWAΨUm 5 CUG > p] 3− .

    Journal: Nucleic Acids Research

    Article Title: An analytical platform for mass spectrometry-based identification and chemical analysis of RNA in ribonucleoprotein complexes

    doi: 10.1093/nar/gkp732

    Figure Lengend Snippet: LC-MS analysis of RNaseT1 digests of yeast tRNA Phe−1 . ( a ) Base peak chromatogram of the RNase T1 digest of the yeast tRNA Phe−1 preparation (200 fmol). Chromatography was performed as described in the ‘Materials and Methods’ section. Sixteen major oligoribonucleotide peaks, indicated by arrows with peak numbers, are assigned to the fragments of yeast tRNA Phe−1 , tRNA Phe−2 , tRNA Lys−2 , or tRNA Tyr (see Table 1 ). ( b and c ) A typical MS spectrum of the RNase T1 digest of tRNA Phe−1 ; (b) [AUUUAm 2 G > p] 2− and (c) [ACmUGmAAyWAΨUm 5 CUG > p] 3− .

    Article Snippet: RNase T1 was purchased from Worthington (Lakewood, NJ) and further purified by reversed-phase liquid chromatography (RPLC) before use.

    Techniques: Liquid Chromatography with Mass Spectroscopy, Chromatography, Mass Spectrometry

    CID-MS/MS spectrum of [AUUUAm 2 G] 2− derived from RNase T1 digestion of yeast tRNA Phe−1 . The doubly charged ion with m / z = 966.6 was analyzed by CID. The nucleotide sequence was verified by manual interpretation of the a- and c-type (normal text) and w - and y -type ( italic text ) product ion series as indicated in the figure. The parent ion losing methyl guanine [P−B(mG) 2− ], the parent ion losing adenine [P−B(A) 2− ], the y5 ion losing methyl guanine [y5−B(mG) 2− ], the y5 ion losing adenine [y5−B(A) 2− ], y5 2− and c5 2− were doubly charged products. All other assigned signals were singly charged products, unless indicated otherwise. The asterisks indicate hydrated or dehydrated ions of a-, c-, w- or y-type products.

    Journal: Nucleic Acids Research

    Article Title: An analytical platform for mass spectrometry-based identification and chemical analysis of RNA in ribonucleoprotein complexes

    doi: 10.1093/nar/gkp732

    Figure Lengend Snippet: CID-MS/MS spectrum of [AUUUAm 2 G] 2− derived from RNase T1 digestion of yeast tRNA Phe−1 . The doubly charged ion with m / z = 966.6 was analyzed by CID. The nucleotide sequence was verified by manual interpretation of the a- and c-type (normal text) and w - and y -type ( italic text ) product ion series as indicated in the figure. The parent ion losing methyl guanine [P−B(mG) 2− ], the parent ion losing adenine [P−B(A) 2− ], the y5 ion losing methyl guanine [y5−B(mG) 2− ], the y5 ion losing adenine [y5−B(A) 2− ], y5 2− and c5 2− were doubly charged products. All other assigned signals were singly charged products, unless indicated otherwise. The asterisks indicate hydrated or dehydrated ions of a-, c-, w- or y-type products.

    Article Snippet: RNase T1 was purchased from Worthington (Lakewood, NJ) and further purified by reversed-phase liquid chromatography (RPLC) before use.

    Techniques: Mass Spectrometry, Derivative Assay, Sequencing

    Base peak chromatogram of the RNaseT1 digest of yeast U4 snRNA isolated from the Lsm3-associated RNP complex. The gel piece containing U4 snRNA was in-gel digested with RNaseT1 and subjected to the LC-MS analysis. Major oligoribonucleotide peaks assigned as RNase T1 fragments of yeast U4 snRNA are indicated by arrows with the corresponding sequence. Detailed data for MS/MS-based assignment of each fragment are given in Supplementary Table S3 .

    Journal: Nucleic Acids Research

    Article Title: An analytical platform for mass spectrometry-based identification and chemical analysis of RNA in ribonucleoprotein complexes

    doi: 10.1093/nar/gkp732

    Figure Lengend Snippet: Base peak chromatogram of the RNaseT1 digest of yeast U4 snRNA isolated from the Lsm3-associated RNP complex. The gel piece containing U4 snRNA was in-gel digested with RNaseT1 and subjected to the LC-MS analysis. Major oligoribonucleotide peaks assigned as RNase T1 fragments of yeast U4 snRNA are indicated by arrows with the corresponding sequence. Detailed data for MS/MS-based assignment of each fragment are given in Supplementary Table S3 .

    Article Snippet: RNase T1 was purchased from Worthington (Lakewood, NJ) and further purified by reversed-phase liquid chromatography (RPLC) before use.

    Techniques: Isolation, Liquid Chromatography with Mass Spectroscopy, Sequencing, Mass Spectrometry