dispase  (Worthington Biochemical)


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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
    Dispase, supplied by Worthington Biochemical, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    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:

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    Worthington Biochemical neonatal cardiomyocytes isolation system
    Overexpression of Tsg101 enhances recycling of IGF-1R in <t>cardiomyocytes.</t> A , B ) Representative immunoblots ( A ) and quantitative analyses ( B ) of streptavidin immunoprecipitates (IP), visualized using an IGF-1R antibody. Whole-cell lysates (WCLs) were used as loading controls. Four independent experiments were performed for recycling assays. C ) Representative images of immunofluorescence staining with streptavidin Alexa Fluor 594 conjugate after IGF-1R biotin was internalized and recycled in Ad.Tsg101 and Ad.GFP NRCMs; n = 4 plates, 25–30 cells per plate. Scale bars, 10 μm. * P
    Neonatal Cardiomyocytes Isolation System, 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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    97
    Worthington Biochemical collagenase type 2
    Adipose tissue immune remodeling during WL and WC. Body weight gain results in a transition from abundant <t>type</t> 2 immune cells present in lean adipose tissue towards an accumulation of immune cells/phenotypes associated with lipid handling (lipid associated macrophages, LAMs) and antigen presentation. WL does not restore the antigen presentation phenotype and is associated with T cell exhaustion. T cell exhaustion and LAMs are further amplified by weight regain. There is a di sconnect between systemic glucose homeostasis and immune remodeling in adipose tissue, such that WL corrects obesity-induced glucose intolerance, but does not resolve the altered immune cell composition caused by diet-induced obesity.
    Collagenase Type 2, 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
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    96
    Worthington Biochemical collagenase type iii
    Chemerin release and NK cell anti-tumour defense account for the improved growth restriction on cisplatin treatment in Mut (LysMCre/VEGF f / f ) mice. ( a ) Percentage of NK1.1-positive cells in cisplatin-treated tumours from WT and Mut mice that received i.p. injections of PBS as control (upper panel) or chemerin-neutralizing antibody (lower panel), determined by flow <t>cytometry.</t> Tumours were analysed in <t>three</t> independent experiments. ( b ) Representative micrographs of SA-β-gal-activity at pH 6.0 on day 18 in tumour sections after treatment with CDDP alone (upper panel), with CDDP and chemerin-neutralizing antibody (middle panel) or with CDDP and NK-depleting antibody mAB PK136 (lower panel). ( c ) Quantification of SA-β-gal-positive cells in b (untreated, n ≥4; CDDP, n ≥6). ( d ) Tumour volumes on day 18 after treatment with CDDP alone, CDDP and chemerin-neutralizing antibody, CDDP and mAB PK136 or CDDP and intratumoural injections of recombinant chemerin ( n ≥5). Bars represent mean values; error bars indicate the s.e.m.; statistical significance was determined by one-way analysis of variance followed by Bonferroni post-hoc test when more than two groups were compared. Statistical significance is indicated as * P
    Collagenase Type Iii, supplied by Worthington Biochemical, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    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: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Overexpression of Tsg101 enhances recycling of IGF-1R in cardiomyocytes. A , B ) Representative immunoblots ( A ) and quantitative analyses ( B ) of streptavidin immunoprecipitates (IP), visualized using an IGF-1R antibody. Whole-cell lysates (WCLs) were used as loading controls. Four independent experiments were performed for recycling assays. C ) Representative images of immunofluorescence staining with streptavidin Alexa Fluor 594 conjugate after IGF-1R biotin was internalized and recycled in Ad.Tsg101 and Ad.GFP NRCMs; n = 4 plates, 25–30 cells per plate. Scale bars, 10 μm. * P

    Journal: The FASEB Journal

    Article Title: Tsg101 positively regulates physiologic-like cardiac hypertrophy through FIP3-mediated endosomal recycling of IGF-1R

    doi: 10.1096/fj.201802338RR

    Figure Lengend Snippet: Overexpression of Tsg101 enhances recycling of IGF-1R in cardiomyocytes. A , B ) Representative immunoblots ( A ) and quantitative analyses ( B ) of streptavidin immunoprecipitates (IP), visualized using an IGF-1R antibody. Whole-cell lysates (WCLs) were used as loading controls. Four independent experiments were performed for recycling assays. C ) Representative images of immunofluorescence staining with streptavidin Alexa Fluor 594 conjugate after IGF-1R biotin was internalized and recycled in Ad.Tsg101 and Ad.GFP NRCMs; n = 4 plates, 25–30 cells per plate. Scale bars, 10 μm. * P

    Article Snippet: NRCMs were isolated using the Worthington Neonatal Cardiomyocytes Isolation System (Worthington Biochemical) according to the manufacturer’s procedures.

    Techniques: Over Expression, Western Blot, Immunofluorescence, Staining

    Overexpression of Tsg101 in NRCMs promotes cell hypertrophy. A ) Diagrams depicting the recombinant adenoviral vectors Ad.GFP and Ad.Tsg101. B ) Representative images showing Ad.GFP- or Ad.Tsg101-infected NRCMs (left panel is under bright field; right panel is the same field under fluorescence microscope). Scale bars, 20 μm. C ) Western blot showing increased expression of Tsg101 in Ad.Tsg101 cardiomyocytes compared with Ad.GFP cells. D , E ) Measurement of myocyte cross-sectional area in those infected cells (green) that were stained with cardiomyocyte-specific α-actinin antibody (red). Scale bars, 20 μm. n = 6 plates, 25–30 cells per plate. F , G ) Representative immunoblots ( F ) and quantitative analyses ( G ) showing expression of PM IGF-1R, total IGF-1R, Akt phosphorylation at Ser473, Rab5a, Rab4a, Rab7, Rab11a, and FIP3 in Ad.Tsg101 and Ad.GFP cells. GAPDH was used as loading control for total protein, total Akt was loading control for Akt phosphorylation, and Na/K-ATPase was loading control for PM protein. PM, plasma membrane; n = 4 for independent experiments. * P

    Journal: The FASEB Journal

    Article Title: Tsg101 positively regulates physiologic-like cardiac hypertrophy through FIP3-mediated endosomal recycling of IGF-1R

    doi: 10.1096/fj.201802338RR

    Figure Lengend Snippet: Overexpression of Tsg101 in NRCMs promotes cell hypertrophy. A ) Diagrams depicting the recombinant adenoviral vectors Ad.GFP and Ad.Tsg101. B ) Representative images showing Ad.GFP- or Ad.Tsg101-infected NRCMs (left panel is under bright field; right panel is the same field under fluorescence microscope). Scale bars, 20 μm. C ) Western blot showing increased expression of Tsg101 in Ad.Tsg101 cardiomyocytes compared with Ad.GFP cells. D , E ) Measurement of myocyte cross-sectional area in those infected cells (green) that were stained with cardiomyocyte-specific α-actinin antibody (red). Scale bars, 20 μm. n = 6 plates, 25–30 cells per plate. F , G ) Representative immunoblots ( F ) and quantitative analyses ( G ) showing expression of PM IGF-1R, total IGF-1R, Akt phosphorylation at Ser473, Rab5a, Rab4a, Rab7, Rab11a, and FIP3 in Ad.Tsg101 and Ad.GFP cells. GAPDH was used as loading control for total protein, total Akt was loading control for Akt phosphorylation, and Na/K-ATPase was loading control for PM protein. PM, plasma membrane; n = 4 for independent experiments. * P

    Article Snippet: NRCMs were isolated using the Worthington Neonatal Cardiomyocytes Isolation System (Worthington Biochemical) according to the manufacturer’s procedures.

    Techniques: Over Expression, Recombinant, Infection, Fluorescence, Microscopy, Western Blot, Expressing, Staining

    Tsg101-mediated cardiomyocyte hypertrophy is attenuated by monensin and KD of FIP3. A , B ) Treatment of NRCMs with monensin (1 μM) 1 h prior to adenovirus infection resulted in reduced plasma membrane levels of IGF-1R; n = 4 for independent experiments; * P

    Journal: The FASEB Journal

    Article Title: Tsg101 positively regulates physiologic-like cardiac hypertrophy through FIP3-mediated endosomal recycling of IGF-1R

    doi: 10.1096/fj.201802338RR

    Figure Lengend Snippet: Tsg101-mediated cardiomyocyte hypertrophy is attenuated by monensin and KD of FIP3. A , B ) Treatment of NRCMs with monensin (1 μM) 1 h prior to adenovirus infection resulted in reduced plasma membrane levels of IGF-1R; n = 4 for independent experiments; * P

    Article Snippet: NRCMs were isolated using the Worthington Neonatal Cardiomyocytes Isolation System (Worthington Biochemical) according to the manufacturer’s procedures.

    Techniques: Infection

    Diagram showing structural components and major players involved in the Tsg101-mediated endosomal recycling of IGF-1R. Tsg101 interacts with and enhances the function of FIP3 at the ERC. Increased activity of the ERC promotes translocation of IGF-1R to the plasma membrane, which in turn augments IGF-IR and Akt signaling in cardiac myocytes, leading to physiologic cardiac hypertrophy.

    Journal: The FASEB Journal

    Article Title: Tsg101 positively regulates physiologic-like cardiac hypertrophy through FIP3-mediated endosomal recycling of IGF-1R

    doi: 10.1096/fj.201802338RR

    Figure Lengend Snippet: Diagram showing structural components and major players involved in the Tsg101-mediated endosomal recycling of IGF-1R. Tsg101 interacts with and enhances the function of FIP3 at the ERC. Increased activity of the ERC promotes translocation of IGF-1R to the plasma membrane, which in turn augments IGF-IR and Akt signaling in cardiac myocytes, leading to physiologic cardiac hypertrophy.

    Article Snippet: NRCMs were isolated using the Worthington Neonatal Cardiomyocytes Isolation System (Worthington Biochemical) according to the manufacturer’s procedures.

    Techniques: Activity Assay, Translocation Assay

    Adipose tissue immune remodeling during WL and WC. Body weight gain results in a transition from abundant type 2 immune cells present in lean adipose tissue towards an accumulation of immune cells/phenotypes associated with lipid handling (lipid associated macrophages, LAMs) and antigen presentation. WL does not restore the antigen presentation phenotype and is associated with T cell exhaustion. T cell exhaustion and LAMs are further amplified by weight regain. There is a di sconnect between systemic glucose homeostasis and immune remodeling in adipose tissue, such that WL corrects obesity-induced glucose intolerance, but does not resolve the altered immune cell composition caused by diet-induced obesity.

    Journal: bioRxiv

    Article Title: Multiomics reveals persistence of obesity-associated immune cell phenotypes in adipose tissue during weight loss and subsequent weight regain

    doi: 10.1101/2021.08.20.455954

    Figure Lengend Snippet: Adipose tissue immune remodeling during WL and WC. Body weight gain results in a transition from abundant type 2 immune cells present in lean adipose tissue towards an accumulation of immune cells/phenotypes associated with lipid handling (lipid associated macrophages, LAMs) and antigen presentation. WL does not restore the antigen presentation phenotype and is associated with T cell exhaustion. T cell exhaustion and LAMs are further amplified by weight regain. There is a di sconnect between systemic glucose homeostasis and immune remodeling in adipose tissue, such that WL corrects obesity-induced glucose intolerance, but does not resolve the altered immune cell composition caused by diet-induced obesity.

    Article Snippet: Stromal Vascular Fraction Isolation and Cell Sorting Mice were euthanized by isoflurane overdose and cervical dislocation and perfused with 20 ml PBS through the left ventricle. eAT pads were collected, minced, and digested in 6 ml of 2-mg/mL type II collagenase (Worthington # LS004177) for 30 min at 37°C.

    Techniques: Amplification

    Chemerin release and NK cell anti-tumour defense account for the improved growth restriction on cisplatin treatment in Mut (LysMCre/VEGF f / f ) mice. ( a ) Percentage of NK1.1-positive cells in cisplatin-treated tumours from WT and Mut mice that received i.p. injections of PBS as control (upper panel) or chemerin-neutralizing antibody (lower panel), determined by flow cytometry. Tumours were analysed in three independent experiments. ( b ) Representative micrographs of SA-β-gal-activity at pH 6.0 on day 18 in tumour sections after treatment with CDDP alone (upper panel), with CDDP and chemerin-neutralizing antibody (middle panel) or with CDDP and NK-depleting antibody mAB PK136 (lower panel). ( c ) Quantification of SA-β-gal-positive cells in b (untreated, n ≥4; CDDP, n ≥6). ( d ) Tumour volumes on day 18 after treatment with CDDP alone, CDDP and chemerin-neutralizing antibody, CDDP and mAB PK136 or CDDP and intratumoural injections of recombinant chemerin ( n ≥5). Bars represent mean values; error bars indicate the s.e.m.; statistical significance was determined by one-way analysis of variance followed by Bonferroni post-hoc test when more than two groups were compared. Statistical significance is indicated as * P

    Journal: Nature Communications

    Article Title: Targeting VEGF-A in myeloid cells enhances natural killer cell responses to chemotherapy and ameliorates cachexia

    doi: 10.1038/ncomms12528

    Figure Lengend Snippet: Chemerin release and NK cell anti-tumour defense account for the improved growth restriction on cisplatin treatment in Mut (LysMCre/VEGF f / f ) mice. ( a ) Percentage of NK1.1-positive cells in cisplatin-treated tumours from WT and Mut mice that received i.p. injections of PBS as control (upper panel) or chemerin-neutralizing antibody (lower panel), determined by flow cytometry. Tumours were analysed in three independent experiments. ( b ) Representative micrographs of SA-β-gal-activity at pH 6.0 on day 18 in tumour sections after treatment with CDDP alone (upper panel), with CDDP and chemerin-neutralizing antibody (middle panel) or with CDDP and NK-depleting antibody mAB PK136 (lower panel). ( c ) Quantification of SA-β-gal-positive cells in b (untreated, n ≥4; CDDP, n ≥6). ( d ) Tumour volumes on day 18 after treatment with CDDP alone, CDDP and chemerin-neutralizing antibody, CDDP and mAB PK136 or CDDP and intratumoural injections of recombinant chemerin ( n ≥5). Bars represent mean values; error bars indicate the s.e.m.; statistical significance was determined by one-way analysis of variance followed by Bonferroni post-hoc test when more than two groups were compared. Statistical significance is indicated as * P

    Article Snippet: Flow cytometry analysis Tumours were digested with 0.1% collagenase type III (Worthington) and 1 U ml−1 DNAse I (Promega) and incubated at 37 °C for 1 h. One hundred and six tumour cells were incubated with Fc-Block (BD Biosciences, #553141 at 1/100) before labelling with fluorochrome-conjugated antibodies.

    Techniques: Mouse Assay, Flow Cytometry, Cytometry, Activity Assay, Recombinant

    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