Here a fluorescent probe based on a carbazole derivative(CNS)was developed to increase the detection range and reduce the detection limit of brilliant blue.Characteristics of CNS are studied.Due to the quenching abili...Here a fluorescent probe based on a carbazole derivative(CNS)was developed to increase the detection range and reduce the detection limit of brilliant blue.Characteristics of CNS are studied.Due to the quenching ability of colorants,CNS shows an excellent current response to brilliant blue(from 1 to 10μM)with a detection limit of 2.7×10^(-8)mol/L(3σ/k)in the conditions of a 1:1 volume ratio of water to tetrahydrofuran.And the stability and reproducibility of CNS in the detection of actual samples indicate great potential for application.展开更多
[Objective] This study aimed to investigate the effects of temperature on determination of protein concentration with Coomassie Brilliant Blue method,thus proving advice and guidance for accurate determination of prot...[Objective] This study aimed to investigate the effects of temperature on determination of protein concentration with Coomassie Brilliant Blue method,thus proving advice and guidance for accurate determination of protein concentration.[Method] With Coomassie Brilliant Blue method,the concentrations of different bovine serum albumin samples were determined under different temperatures and incubation time.[Result] According to the standard curve,when the determination range of protein concentration was 0-100 mg/ml,the determined protein concentration was relatively stable after incubation at 20 ℃ for 20-30 min.Furthermore,the determination result of higher protein concentration with Coomassie Brilliant Blue method was less affected by various factors.[Conclusion] In determination of protein concentration with Coomassie Brilliant Blue method,temperature,sample concentration and incubation time were important factors affecting the accuracy of experimental results.展开更多
Aspergillus ficuum was immobilized with sodium alginate, and decolourization of Reactive Brilliant Blue KN-R was studied on immobilized and free Aspergillus ficuum. The optimal preparation condition of the strain immo...Aspergillus ficuum was immobilized with sodium alginate, and decolourization of Reactive Brilliant Blue KN-R was studied on immobilized and free Aspergillus ficuum. The optimal preparation condition of the strain immobilization was obtained by the orthogonal test, it is sodium alginate 3%, CaCl_2 5%, wet mycelia 30 g/L, calcific time 8 h. It was found that the immobilized cells could effectively decolourize Reactive Brilliant Blue KN-R, the optimum temperature and pH were 33℃ and 5.0, respectively. The kinetics study of decolourization of immobilized cells showed that the decolourization of Aspergillus ficuum immobilized conformed to zero-order reaction model. The decolourization efficiency of immobilized cell compared with that of free cell in different physical conditions. Results showed that the decolourization of immobilized cells with mycelia had the best efficiency. The immobilized cells could be reused after the first decolourization.展开更多
Previous studies have confirmed that oxidized adenosine triphosphate, a P2X7 receptor antagonist, attenuates lipopolysaccharide-mediated microglial activation and inflammatory expression following neuronal damage in r...Previous studies have confirmed that oxidized adenosine triphosphate, a P2X7 receptor antagonist, attenuates lipopolysaccharide-mediated microglial activation and inflammatory expression following neuronal damage in rat brain. NaCI and temperature may affect the potency of oxidized adenosine triphosphate. Brilliant blue G is a derivative of a widely used food additive and has little toxicity. This study explored the effects of brilliant blue G, a selective P2X7 receptor antagonist, on microglial activation and inflammation. Results demonstrated that brilliant blue G inhibited the release of cydooxygenase-2 and interleukin-6 in BV2 cells. Immunofluorescence displayed that brilliant blue G could suppress lipopolysaccharide-induced microglial activation. This study used RNA interference to block P2X7 receptor expression and found that small interfering RNA also suppressed the release of cyclooxygenase-2 and interleukin-6 in BV2 cells. These results suggested that downregulation of the P2X7 receptor by brilliant blue G was involved in the inhibition of microglial activation and inflammation.展开更多
Differentiating pasteurized milk and reconsti-tuted milk by scientific approach was necessary to defend consumer from economic fraud of wrong labeling. In this paper 2DGE (2 Dimen-sional Gel Electrophoresis)-coomassie...Differentiating pasteurized milk and reconsti-tuted milk by scientific approach was necessary to defend consumer from economic fraud of wrong labeling. In this paper 2DGE (2 Dimen-sional Gel Electrophoresis)-coomassie brilliant blue staining method was employed and sig-nificant color intensity changing was observed among raw milk, pasteurized milk, UHT milk and reconstituted milk. For example, the intensity of 10 protein spots including casein and lac-toglobulin reduced more than two folds from pasteurized milk to reconstituted milk. However, DIGE (Differential Gel Electrophoresis) assay showed that the majority protein remained simi-lar level from pasteurized milk to reconstituted milk. Therefore the color fading of coomassie brilliant blue stained 2D gels may be due to other biochemical reaction, such as Maillard reaction, instead of protein degradation. Stability of 2DGE pattern was confirmed by running six gels of the same sample in parallel and software analysis showed that all proteins were at similar level. Two commercialized pasteurized milk samples and one reconstituted milk sample were tested by 2DGE-coomassie blue staining method and re-constituted milk could be easily identified.展开更多
An improved CBB staining with higher sensitivity than that of the typical CBB staining was reported.The main improvement was using a fixing step of 25% trichloroacetic acid(TCA) before CBB staining.For most proteins s...An improved CBB staining with higher sensitivity than that of the typical CBB staining was reported.The main improvement was using a fixing step of 25% trichloroacetic acid(TCA) before CBB staining.For most proteins studied,the sensitivity of the improved CBB staining was about twice as high as that of the typical method.For basic and low molecular weight proteins such as ribosomal proteins,the sensitivity of this improved staining method was about 3.5-28 times that of the typical method.It was speculated that the improved procedure would be suitable for exact quantitative analysis of proteins fractionated by SDS-PAGE,especially for basic and low molecular weight proteins.On the other hand,this new modified method might be also applied to multidisciplinary studies,such as biological researches and nuclear sciences.展开更多
A study of the decolorization of reactive brilliant blue in an aqueous solution using Fe-Mn-sepiolite as a heterogeneous Fenton-like catalyst has been performed. The Fourier transform infrared （FTIR） spectra of the ...A study of the decolorization of reactive brilliant blue in an aqueous solution using Fe-Mn-sepiolite as a heterogeneous Fenton-like catalyst has been performed. The Fourier transform infrared （FTIR） spectra of the catalyst showed bending vibrations of the Fe-O. The Xray diffraction （XRD） patterns of the catalyst showed characteristic diffraction peaks of α-Fe203, γ-Fe203 and MnO. A four factor central composite design （CCD） coupled with response surface methodology （RSM） was applied to evaluate and optimize the important variables （catalyst addition, hydrogen peroxide dosage, initial pH value and initial dye concentration）. When the reaction conditions were catalyst dosage = 0.4 g, [H202]= 0.3 mL, pH= 2.5, [reactive brilliant blue]o = 50mg·L-1, and volume of solution = 500 mL at room temperature, the decolorization efficiency of reactive brilliant blue was 91.98% within 60min. Moreover, the Fe-Mn-sepiolite catalyst had good stability for the degradation of reactive brilliant blue even after six cycles. Leaching of iron ions （ 〈 0.4 mg·L-l） was observed. The decoloring process was reactive brilliant blue specific via a redox reaction. The benzene ring and naphthalene ring were first oxidized to open ring; these were then oxidized to the alcohol and carboxylic acid. The reactive brilliant blue was decom- posed mainly by the attack of .OH radicals including surface-bound .OH radicals generated on the catalyst surface.展开更多
Fungi play an important role in dying wastewater treatment.In this work,the mycelia of Lactarius deliciosus exhibited an excellent capacity in decolorizing coomassie brilliant blue(CBB).The results demonstrated that t...Fungi play an important role in dying wastewater treatment.In this work,the mycelia of Lactarius deliciosus exhibited an excellent capacity in decolorizing coomassie brilliant blue(CBB).The results demonstrated that the mycelia could treat CBB with high concentrations over a broad range of pH and temperature.The decolorization rate of 99.19%and the removal rate of 16.31 mg·L^(‒1)·h were realized.The mycelia could be recycled from decolorizing process for 19 times,indicating a good re-usability.It verified that the lignin peroxidase(121.65 U·L^(‒1))and manganese peroxidase(36.77 U·L^(‒1))were involved in the degradation and decolorization process of CBB.Toxicity assessments indicated the seed germination rate was up to 82.22%while inhibition to Escherichia coli decreased dramatically and no significant effect on Caenorhabditis elegans growth was found.The removal of CBB was a synergistic process accomplished by adsorption and biodegradation.The mycelia could be used for eco-friendly CBB treatment.展开更多
[Objective] The aim of this study was to increase the viability of sheep oocytes in vitro by using phosphodiesterase type 3（PDE 3） inhibitor milrinone combined with brilliant cresyl blue（BCB） staining.[Method] The...[Objective] The aim of this study was to increase the viability of sheep oocytes in vitro by using phosphodiesterase type 3（PDE 3） inhibitor milrinone combined with brilliant cresyl blue（BCB） staining.[Method] The differences between BCB tested and morphologically selected oocytes,as well as the effect of them on embryo development were compared;and then suitable inhibitive time of milrinone to sheep oocytes in vitro was studied and used in BCB-oocytes for in vitro embryo production（IVEP）.[Result] The BCB＋ oocytes percentage in A-and B-level sheep oocytes was 64.42%,which was extremely significantly higher than that in C-level（17.0%）.The maturing rate,cleavage rate and blastocyst rate of BCB＋ oocytes（86.16%,85.29% and 34.40%） of was significantly higher than those of BCB-oocytes（50.94%,36.19% and 6.73%）.The best time for PDE 3 inhibitor delaying the sheep oocyte mature in vitro was 6 h.In addition,the rate of embryo development in vitro could be significantly increased by inhibiting the BCB-oocytes for 6 h with Milrinone.[Conclusion] The study will provide reference for improving the efficiency of sheep oocytes culture in vitro.展开更多
Selecting oocytes that are most likely to develop is crucial for in vitro fertilization and animal cloning. Brilliant cresyl blue （BCB） staining has been used for oocyte selection in large animals, but its wider uti...Selecting oocytes that are most likely to develop is crucial for in vitro fertilization and animal cloning. Brilliant cresyl blue （BCB） staining has been used for oocyte selection in large animals, but its wider utility needs further evaluation. Mouse oocytes were divided into those stained （BCB＋） and those unstained （BCB-） according to their ooplasm BCB coloration. Chromatin configurations, cumulus cell apoptosis, cytoplasmic maturity and developmental competence were compared between the BCB＋ and BCB- oocytes. The effects of oocyte diameter, sexual maturity and gonadotropin stimulation on the competence of BCB＋ oocytes were also analyzed. In the large- and medium-size groups, BCB＋ oocytes were larger and showed more surrounded nucleoli （SN） chromatin configurations and higher frequencies of early atresia, and they also gained better cytoplasmic maturity （determined as the intracellular GSH level and pattern of mitochondrial distribution） and higher developmental potential after in vitro maturation （IVM） than the BCB-oocytes. Adult mice produced more BCB＋ oocytes with higher competence than the prepubertal mice when not primed with PMSG. PMSG priming increased both proportion and developmental potency of BCB＋ oocytes. The BCB＋ oocytes in the large-size group showed more SN chromatin configurations, better cytoplasmic maturity and higher developmental potential than their counterparts in the medium-size group. It is concluded that BCB staining can be used as an efficient method for oocyte selection, but that the competence of the BCB＋ oocytes may vary with oocyte diameter, animal sexual maturity and gonadotropin stimulation. Taken together, the series of criteria described here would allow for better choices in selecting oocytes for better development.展开更多
Heparin is a polysaccharide of glycosaminoglycan class, which consists of repeating disaccharide units of iduronic/glucuronic acid and glucosamine residues with many biological functions. Many methods have been propos...Heparin is a polysaccharide of glycosaminoglycan class, which consists of repeating disaccharide units of iduronic/glucuronic acid and glucosamine residues with many biological functions. Many methods have been proposed for the detection of heparin, including UV-Vis spectrophotometry, the light scattering technique, HPLC and electro phoresis and flow injection analysis, etc.. But there are few reports about the detection of heparin by means of an electrochemical method. Electroanalytical methods are useful tools in bioanalytical chemistry because of their advantages, such as their instrumental simplicity, moderate cost and portability. The binding reactions of organic molecules with biomolecules such as DNA and proteins have been widely studied. In acidic solution, heparin is highly negatively charged due to the dissociation of the sulfate and carboxyl groups in its molecule, which can easily interact with cationic dyes. Based on this principle, in this work, a new electrochemical method for the determination of heparin was developed based on the interaction of heparin with brilliant cresyl blue（BCB）.展开更多
A simplified method is presented for tryptic digestion of Coomassie brilliant blue(CBB)-stained proteins in polyacrylamide gels. Compared with conventional methods, the proposed method does not require a removal of th...A simplified method is presented for tryptic digestion of Coomassie brilliant blue(CBB)-stained proteins in polyacrylamide gels. Compared with conventional methods, the proposed method does not require a removal of the dye before digestion, and is thus faster and saves a lot of labor. The resulted digest can be analyzed by either RPLC/ESIMS or MALDI MS for identification of the protein in a conventional way. Model studies with bovine serum albumin(BSA) showed that 50 ng of the protein could be routinely identified. The simplified procedure displays a tendency to produce more incompletely cleaved peptides, which is favorable for improving the sequence coverage.展开更多
基金Funded by the Open Subject from Jiangsu Marine Resources Development Research Institute (JSIMR202117)the Training Programs of Innovation and Entrepreneurship for College Students in Jiangsu Ocean University。
文摘Here a fluorescent probe based on a carbazole derivative(CNS)was developed to increase the detection range and reduce the detection limit of brilliant blue.Characteristics of CNS are studied.Due to the quenching ability of colorants,CNS shows an excellent current response to brilliant blue(from 1 to 10μM)with a detection limit of 2.7×10^(-8)mol/L(3σ/k)in the conditions of a 1:1 volume ratio of water to tetrahydrofuran.And the stability and reproducibility of CNS in the detection of actual samples indicate great potential for application.
基金Supported by Natural Science Foundation of Jilin Province(201115221)~~
文摘[Objective] This study aimed to investigate the effects of temperature on determination of protein concentration with Coomassie Brilliant Blue method,thus proving advice and guidance for accurate determination of protein concentration.[Method] With Coomassie Brilliant Blue method,the concentrations of different bovine serum albumin samples were determined under different temperatures and incubation time.[Result] According to the standard curve,when the determination range of protein concentration was 0-100 mg/ml,the determined protein concentration was relatively stable after incubation at 20 ℃ for 20-30 min.Furthermore,the determination result of higher protein concentration with Coomassie Brilliant Blue method was less affected by various factors.[Conclusion] In determination of protein concentration with Coomassie Brilliant Blue method,temperature,sample concentration and incubation time were important factors affecting the accuracy of experimental results.
文摘Aspergillus ficuum was immobilized with sodium alginate, and decolourization of Reactive Brilliant Blue KN-R was studied on immobilized and free Aspergillus ficuum. The optimal preparation condition of the strain immobilization was obtained by the orthogonal test, it is sodium alginate 3%, CaCl_2 5%, wet mycelia 30 g/L, calcific time 8 h. It was found that the immobilized cells could effectively decolourize Reactive Brilliant Blue KN-R, the optimum temperature and pH were 33℃ and 5.0, respectively. The kinetics study of decolourization of immobilized cells showed that the decolourization of Aspergillus ficuum immobilized conformed to zero-order reaction model. The decolourization efficiency of immobilized cell compared with that of free cell in different physical conditions. Results showed that the decolourization of immobilized cells with mycelia had the best efficiency. The immobilized cells could be reused after the first decolourization.
基金supported by the National Natural Science Foundation of China, No. 81072242the Excellent Supervisor & Yat-Sen Creative Talent Development Program of Sun Yat-sen University
文摘Previous studies have confirmed that oxidized adenosine triphosphate, a P2X7 receptor antagonist, attenuates lipopolysaccharide-mediated microglial activation and inflammatory expression following neuronal damage in rat brain. NaCI and temperature may affect the potency of oxidized adenosine triphosphate. Brilliant blue G is a derivative of a widely used food additive and has little toxicity. This study explored the effects of brilliant blue G, a selective P2X7 receptor antagonist, on microglial activation and inflammation. Results demonstrated that brilliant blue G inhibited the release of cydooxygenase-2 and interleukin-6 in BV2 cells. Immunofluorescence displayed that brilliant blue G could suppress lipopolysaccharide-induced microglial activation. This study used RNA interference to block P2X7 receptor expression and found that small interfering RNA also suppressed the release of cyclooxygenase-2 and interleukin-6 in BV2 cells. These results suggested that downregulation of the P2X7 receptor by brilliant blue G was involved in the inhibition of microglial activation and inflammation.
文摘Differentiating pasteurized milk and reconsti-tuted milk by scientific approach was necessary to defend consumer from economic fraud of wrong labeling. In this paper 2DGE (2 Dimen-sional Gel Electrophoresis)-coomassie brilliant blue staining method was employed and sig-nificant color intensity changing was observed among raw milk, pasteurized milk, UHT milk and reconstituted milk. For example, the intensity of 10 protein spots including casein and lac-toglobulin reduced more than two folds from pasteurized milk to reconstituted milk. However, DIGE (Differential Gel Electrophoresis) assay showed that the majority protein remained simi-lar level from pasteurized milk to reconstituted milk. Therefore the color fading of coomassie brilliant blue stained 2D gels may be due to other biochemical reaction, such as Maillard reaction, instead of protein degradation. Stability of 2DGE pattern was confirmed by running six gels of the same sample in parallel and software analysis showed that all proteins were at similar level. Two commercialized pasteurized milk samples and one reconstituted milk sample were tested by 2DGE-coomassie blue staining method and re-constituted milk could be easily identified.
文摘An improved CBB staining with higher sensitivity than that of the typical CBB staining was reported.The main improvement was using a fixing step of 25% trichloroacetic acid(TCA) before CBB staining.For most proteins studied,the sensitivity of the improved CBB staining was about twice as high as that of the typical method.For basic and low molecular weight proteins such as ribosomal proteins,the sensitivity of this improved staining method was about 3.5-28 times that of the typical method.It was speculated that the improved procedure would be suitable for exact quantitative analysis of proteins fractionated by SDS-PAGE,especially for basic and low molecular weight proteins.On the other hand,this new modified method might be also applied to multidisciplinary studies,such as biological researches and nuclear sciences.
文摘A study of the decolorization of reactive brilliant blue in an aqueous solution using Fe-Mn-sepiolite as a heterogeneous Fenton-like catalyst has been performed. The Fourier transform infrared （FTIR） spectra of the catalyst showed bending vibrations of the Fe-O. The Xray diffraction （XRD） patterns of the catalyst showed characteristic diffraction peaks of α-Fe203, γ-Fe203 and MnO. A four factor central composite design （CCD） coupled with response surface methodology （RSM） was applied to evaluate and optimize the important variables （catalyst addition, hydrogen peroxide dosage, initial pH value and initial dye concentration）. When the reaction conditions were catalyst dosage = 0.4 g, [H202]= 0.3 mL, pH= 2.5, [reactive brilliant blue]o = 50mg·L-1, and volume of solution = 500 mL at room temperature, the decolorization efficiency of reactive brilliant blue was 91.98% within 60min. Moreover, the Fe-Mn-sepiolite catalyst had good stability for the degradation of reactive brilliant blue even after six cycles. Leaching of iron ions （ 〈 0.4 mg·L-l） was observed. The decoloring process was reactive brilliant blue specific via a redox reaction. The benzene ring and naphthalene ring were first oxidized to open ring; these were then oxidized to the alcohol and carboxylic acid. The reactive brilliant blue was decom- posed mainly by the attack of .OH radicals including surface-bound .OH radicals generated on the catalyst surface.
基金This work was supported by the Anhui Provincial Program on Key Research and Development Project(Grant No.202004a06020021)the National Natural Science Foundation of China(Grant No.21606002)+1 种基金the Natural Science Foundation of Anhui Province(CN)(Grant No.1708085QC64)the Undergraduate Research Training Programs for Innovation(Grant Nos.201910357069,S201910357427).
文摘Fungi play an important role in dying wastewater treatment.In this work,the mycelia of Lactarius deliciosus exhibited an excellent capacity in decolorizing coomassie brilliant blue(CBB).The results demonstrated that the mycelia could treat CBB with high concentrations over a broad range of pH and temperature.The decolorization rate of 99.19%and the removal rate of 16.31 mg·L^(‒1)·h were realized.The mycelia could be recycled from decolorizing process for 19 times,indicating a good re-usability.It verified that the lignin peroxidase(121.65 U·L^(‒1))and manganese peroxidase(36.77 U·L^(‒1))were involved in the degradation and decolorization process of CBB.Toxicity assessments indicated the seed germination rate was up to 82.22%while inhibition to Escherichia coli decreased dramatically and no significant effect on Caenorhabditis elegans growth was found.The removal of CBB was a synergistic process accomplished by adsorption and biodegradation.The mycelia could be used for eco-friendly CBB treatment.
基金Supported by Natural Science Foundation of Xinjiang AutonomousRegion (200821182 )Science and Technology Research andDevelopment Program of Xinjiang Autonomous Region (200841122)+1 种基金Science and Technology Planning Project of Xinjiang AutonomousRegion (200711104)the National Transgenic Major Program~~
文摘[Objective] The aim of this study was to increase the viability of sheep oocytes in vitro by using phosphodiesterase type 3（PDE 3） inhibitor milrinone combined with brilliant cresyl blue（BCB） staining.[Method] The differences between BCB tested and morphologically selected oocytes,as well as the effect of them on embryo development were compared;and then suitable inhibitive time of milrinone to sheep oocytes in vitro was studied and used in BCB-oocytes for in vitro embryo production（IVEP）.[Result] The BCB＋ oocytes percentage in A-and B-level sheep oocytes was 64.42%,which was extremely significantly higher than that in C-level（17.0%）.The maturing rate,cleavage rate and blastocyst rate of BCB＋ oocytes（86.16%,85.29% and 34.40%） of was significantly higher than those of BCB-oocytes（50.94%,36.19% and 6.73%）.The best time for PDE 3 inhibitor delaying the sheep oocyte mature in vitro was 6 h.In addition,the rate of embryo development in vitro could be significantly increased by inhibiting the BCB-oocytes for 6 h with Milrinone.[Conclusion] The study will provide reference for improving the efficiency of sheep oocytes culture in vitro.
基金Acknowledgments This study was supported by grants from the China National Natural Science Foundation （Nos. 30430530 and 30571337） and from the Momentous Research Project of the China Ministry of Science and Technology （No. 2006CB944003）.
文摘Selecting oocytes that are most likely to develop is crucial for in vitro fertilization and animal cloning. Brilliant cresyl blue （BCB） staining has been used for oocyte selection in large animals, but its wider utility needs further evaluation. Mouse oocytes were divided into those stained （BCB＋） and those unstained （BCB-） according to their ooplasm BCB coloration. Chromatin configurations, cumulus cell apoptosis, cytoplasmic maturity and developmental competence were compared between the BCB＋ and BCB- oocytes. The effects of oocyte diameter, sexual maturity and gonadotropin stimulation on the competence of BCB＋ oocytes were also analyzed. In the large- and medium-size groups, BCB＋ oocytes were larger and showed more surrounded nucleoli （SN） chromatin configurations and higher frequencies of early atresia, and they also gained better cytoplasmic maturity （determined as the intracellular GSH level and pattern of mitochondrial distribution） and higher developmental potential after in vitro maturation （IVM） than the BCB-oocytes. Adult mice produced more BCB＋ oocytes with higher competence than the prepubertal mice when not primed with PMSG. PMSG priming increased both proportion and developmental potency of BCB＋ oocytes. The BCB＋ oocytes in the large-size group showed more SN chromatin configurations, better cytoplasmic maturity and higher developmental potential than their counterparts in the medium-size group. It is concluded that BCB staining can be used as an efficient method for oocyte selection, but that the competence of the BCB＋ oocytes may vary with oocyte diameter, animal sexual maturity and gonadotropin stimulation. Taken together, the series of criteria described here would allow for better choices in selecting oocytes for better development.
文摘Heparin is a polysaccharide of glycosaminoglycan class, which consists of repeating disaccharide units of iduronic/glucuronic acid and glucosamine residues with many biological functions. Many methods have been proposed for the detection of heparin, including UV-Vis spectrophotometry, the light scattering technique, HPLC and electro phoresis and flow injection analysis, etc.. But there are few reports about the detection of heparin by means of an electrochemical method. Electroanalytical methods are useful tools in bioanalytical chemistry because of their advantages, such as their instrumental simplicity, moderate cost and portability. The binding reactions of organic molecules with biomolecules such as DNA and proteins have been widely studied. In acidic solution, heparin is highly negatively charged due to the dissociation of the sulfate and carboxyl groups in its molecule, which can easily interact with cationic dyes. Based on this principle, in this work, a new electrochemical method for the determination of heparin was developed based on the interaction of heparin with brilliant cresyl blue（BCB）.
基金Supported by the National BasicResearch Priorities Program me of China( No.0 0 1CB5 10 2 0 2),National High- TechProgramm e of China( No.2 0 0 1AA2 30 31)andShanghai Science and Technology Developing Program me( No.0 1JC14 0 11)
文摘A simplified method is presented for tryptic digestion of Coomassie brilliant blue(CBB)-stained proteins in polyacrylamide gels. Compared with conventional methods, the proposed method does not require a removal of the dye before digestion, and is thus faster and saves a lot of labor. The resulted digest can be analyzed by either RPLC/ESIMS or MALDI MS for identification of the protein in a conventional way. Model studies with bovine serum albumin(BSA) showed that 50 ng of the protein could be routinely identified. The simplified procedure displays a tendency to produce more incompletely cleaved peptides, which is favorable for improving the sequence coverage.
