cardiomyocyte apoptosis in heart development methods and protocols
Tài liệu Cardiovascular Development Methods and Protocols ppt
Ngày tải lên :
12/02/2014, 16:20
... Culture for Studying Heart Development Calvin T Hang and Ching-Pin Chang Quantifying Cardiac Functions in Embryonic and Adult Zebrafish Tiffany Hoage, Yonghe Ding, and Xiaolei Xu ... plunger and inject resin into the heart It is important to steady the syringe and needle while injecting the resin, particularly for smaller embryonic hearts Additional movement of the bevel within ... molecular biology and new available equipments and techniques, and we begin to understand the molecular pathways and cellular interaction in the process of heart induction, rightward looping, chamber...
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in situ detection of dna damage, methods and protocols
Ngày tải lên :
11/04/2014, 09:47
... counterstain Incubate cells in 0.2 µg/mL DAPI, and rinse once in PBS To label fixed samples using both TUNEL and immunofluorescence staining for an antigen of interest, the antibody incubations ... fluorescent staining of cultured cells and tissues is presented thereby allowing investigators to utilize the TUNEL staining procedure with a minimal investment in laboratory reagents and equipment In addition, ... DNA making it ideal for labeling fragments in apoptotic cells The enzyme will also label single stranded DNA molecules containing a 3'-OH and will attach nucleotides to a single-strand nick in DNA...
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methods in molecular biology, v.474. nanostructure design. methods and protocols, 2008, p.268
Ngày tải lên :
04/06/2014, 15:06
... Protocols and Bioinformatics, edited by Svetlana Gerdes and Andrei L Osterman, 2008 415 Innate Immunity, edited by Jonathan Ewbank and Eric Vivier, 2007 414 Apoptosis in Cancer: Methods and Protocols, ... enabling gene therapy and tissue engineering applications (2022) 1.2 Methodology for Creating and Studying Chimeric Proteins Between Fibritin and Triple-Stranded Segments of Fibrous Proteins The ... creating and studying chimeric proteins was first developed from fundamental studies aimed at structural understanding of fibrous proteins In phage T4 fibritin, a 27-amino acid (aa) domain (amino...
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Glycoprotein Methods and Protocols - P1
Ngày tải lên :
08/10/2012, 10:25
... allowing gland material to be obtained from the remaining tissue From: Methods in Molecular Biology, Vol 125: Glycoprotein Methods and Protocols: The Mucins Edited by: A Corfield © Humana Press Inc., ... hexose and sialic acid; membrane-based methods such as slot-blotting and staining with periodic acid-Schiff reagent; antibodies and lectins or coating methods such as the glycan detection method and ... in CsCl/4 M guanidinium chloride, which gives a good separation of higher buoyant density mucins and nucleic acids from low buoyant density proteins, glycoproteins, and lipids while maintaining...
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Glycoprotein Methods and Protocols - P2
Ngày tải lên :
08/10/2012, 10:25
... both mucin and lectin histochemistry 1.2 Interpretation and Reporting of Mucin Staining The interpretation of mucin staining will be incomplete or even misleading if the results are not integrated ... problematic, and particular approaches may be required to distinguish focal but intense staining and diffuse but weak staining Grading of staining intensity is notoriously unreliable in the intermediate ... cultures/cytospins with 0.1% avidin in 0.05 M Tris-HCl followed by 1% D-biotin in 0.05 M Tris-HCl for 10 each with thorough washing in PBS following each incubation 19 Several approaches singly or in combination...
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Glycoprotein Methods and Protocols - P3
Ngày tải lên :
08/10/2012, 10:25
... mucins, in Glycoprotein Methods and Protocols: The Mucins (Corfield, T., ed.), Humana, Totowa, NJ Carraway, K L (2000) Preparation of membrane mucin, in Glycoprotein Methods and Protocols: The Mucins ... Subheading 3.1.1 describes a double-determinant format and Subheading 3.1.2 a competitive binding assay using antibody capture 52 McGuckin and Thornton In this protocol, purified mucin-reactive ... mucins in solution using chemical techniques relies on reactions involving mucin carbohydrate groups and is probably most useful for rapid semiquantitative determination of mucin recovery during...
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Glycoprotein Methods and Protocols - P4
Ngày tải lên :
08/10/2012, 10:25
... Pretreatment in 100% ethanol: Rinse in running tap water: Rinse in 3% (v/v) acetic acid: Stain in 1% Alcian Blue 8GX in 3% acetic acid (pH 2.5): Rinse in 3% acetic acid and then rinse in running tap ... primarily of a fibrin gel and necrotic cells with some mucin staining (15) Original methods for measuring adherent mucous thickness in situ were based on (1) differences in refractive index between ... metabisulfite, repeat three times: 12 Rinse in running tap water: 13 Postfix in paraformaldehyde vapor, at 37°C: 14 Mount in gelatin 20 10 10 min 2.5 h 10 min 15 min min/rinse 45 Notes Removal of the muscle...
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Glycoprotein Methods and Protocols - P5
Ngày tải lên :
08/10/2012, 10:25
... produce mucins (1,4–11) More important, a distinction can be made with the preexisting, stored mucin in the mucin-producing cells Metabolic labeling during short periods (up to 60 min) will not ... Perform all ensuing procedures in Blotto as follows: a Incubate in Blotto for 30 min, and incubate with antibody directed against nonglycosylated peptide epitopes of the mucin of interest for 90 ... radioactivity, owing to the use of the PhosphorImager The identification of mucin precursors and mature mucins using polypeptide-specific antisera and SDS-PAGE is elaborated in Chapters 20 and 21, and in several...
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Glycoprotein Methods and Protocols - P6
Ngày tải lên :
08/10/2012, 10:25
... guanidinium chloride containing proteinase inhibitors and are purified by isopycnic centrifugation in CsCl density gradient first in M guanidinium chloride (removal of proteins) and then in 0.2 ... reverse phase chromatography and individual peptides purified and their primary sequence determined by N-terminal sequencing 3.4.2.1 PEPTIDE PURIFICATION AND N-TERMINAL SEQUENCING Solubilize lyophilized ... Audie, J P., Janin, A., Porchet, N., Copin, M C., Gosselin, B., and Aubert, J P (1993) Expression of human mucin genes in respiratory, digestive, and reproductive tracts ascertained by in situ hybridization...
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Glycoprotein Methods and Protocols - P7
Ngày tải lên :
08/10/2012, 10:25
... stained, see Subheading 3.3.1.3.; if not, go to the next step Wash grid in 95% ethanol, remove excess solution on filter paper, and air-dry 3.3.1.3 STAINING AND SHADOWING Dip grid in staining ... the surface and thereafter washed in 95% ethanol, dried, and rotary shadowed This can also be performed in a microversion by adding the BAC to the mucin in solution and transferring 40-µL drops ... these grids; and the addition of contrast, including positive staining and/ or metal shadowing 3.3.1.1 PREPARATION OF CARBON-COATED GRIDS Prepare a thin carbon film (2–5 nm) by the indirect evaporation...
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Glycoprotein Methods and Protocols - P8
Ngày tải lên :
08/10/2012, 10:25
... threonine, A = alanine, P = proline, Y = tyrosine, R = arginine, V = valine, M = methionine, I = isoleucine, L = leucine, F = phenylalanine, K = lysine the amount or ratio of certain amino acids, so ... mucin amino acid/total mucin amino acids × 100%, where total mucin amino acids is the sum of mucin amino acids (except cysteine and tryptophan; asparagine and glutamine are recovered as their ... Quenching reagent: mL of acetic acid (AR grade) and mL of acetonitrile Amino Acid Analysis of Mucins 115 2.2.2 Chromatography Reagents Amino acid standards H: protein hydrolysate standard in 0.1...
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Glycoprotein Methods and Protocols - P9
Ngày tải lên :
08/10/2012, 10:25
... both macro- and microheterogeneity can be determined for a glycopeptide or glycoprotein in a single experiment (5) This technique has allowed mucin (and mucin-like) glycopeptides and trifluoromethane ... Edman sequencing (Fig 1, vessel 1), and a significant lag is introduced when several prolines occur in a cluster and more than one signal is detected in each cycle 10 Freeze-drying mucins is not ... prior to coupling 16 Incubation at 4°C increases the initial yield (7) 17 To incorporate a methanol extraction into the routine, methanol is placed in an unused bottle position, and solvent delivery...
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Glycoprotein Methods and Protocols - P10
Ngày tải lên :
08/10/2012, 10:25
... precoat the pins in blocking buffer in order to minimize nonspecific binding to the gear To achieve this, immerse the pins in a microtiter plate containing blocking buffer at 200 µL/well and incubate ... pin holders, and reaction trays In addition, it contains all the software needed for creating a synthesis schedule, running dispensing aids, and reading and plotting assay results 3.2.1 Creating ... deprotect and wash the free amino termini as described in step The amino terminus may then be acetylated to remove the charge associated with a free amino terminus (if required) by incubating the pins...
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Glycoprotein Methods and Protocols - P11
Ngày tải lên :
08/10/2012, 10:25
... against synthetic peptides D4553 corresponding to a 14 amino acid segment in the mucin domain, and E20-14, corresponding to the C-terminal 14 amino acids of the mucin domain (21) (see Fig 1) in ... peptides corresponding to the C-terminal domain of intestinal mucins Glycoconj J 13, 81–90 22 Higuchi, R (1990) Recombinant PCR, in PCR Protocols (Innis, M., Gelfand, D H., and Sninsky, J J., eds.), ... Woitach, J T., Davidson, E A., and Bhavanandan, V P (1990) Cloning and cDNA sequence of a bovine submaxillary gland mucin-like protein containing two distinct domains Proc Natl Acad Sci USA 87,...
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Glycoprotein Methods and Protocols - P12
Ngày tải lên :
08/10/2012, 10:25
... chromatographic data by using an integrator After stabilization and the isocratic run, inject 10 µL of a mixture of standards containing 250 pmol of neutral monosaccharides and hexosamines Determine the response ... Fuc; 7, Glc; I.S., internal standard cinnamic acid Monosaccharide Composition of Mucin 175 sodium cyanoborohydride in mL of methanol containing 2-aminopyridine (10% w/v) and acetic acid (10% ... described (see Subheading 3.2.) Evaporate off the solvent and dissolve the residue in 100 µL of coupling reagent (prepared by dissolving 100 mg of 2-aminopyridine in 50 µL of acetic acid and 60 µL of...
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Glycoprotein Methods and Protocols - P13
Ngày tải lên :
08/10/2012, 10:25
... mucin oligosaccharide alditols, containing 320 ng of D-melibiose as internal standard, onto the two CarboPac PA-100 columns (connected in series) equilibrated in eluent A Elute with an increasing ... solution of TFMSA in anisole and cool in dry ice/ethanol Add 10 times the weight of TFMSA in anisole to the lyophilized material in a teflon screw-capped vial standing on a bed of ice Incubate at 0°C ... of Galβ1-3GalNAcα- from mucin using O-Glycanase Reconstitute mucin (100 µg) with 90 µL of 0.1 M citrate-phosphate, pH 6.0, containing 100 µg/mL of bovine serum albumin and 0.02% (w/v) sodium azide...
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Glycoprotein Methods and Protocols - P14
Ngày tải lên :
08/10/2012, 10:25
... structure determination of carbohydrate chains This method was introduced in the 1970s and rapidly received a large application for analyzing the sequence of N-acetyllactosamine- and oligomannosidic-type ... is cleaved in two fragments, one corresponding to the 3-branch and the other to the 6-branch Second, the resulting aldehydes are coupled by reductive amination to 2-aminopyridine: in a first ... included in the review of Kamerling et al (17) clearly shows that a spectrum is unique and can be used as an “identity card.” This review presents the carbohydrate chains in a logical order and...
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Glycoprotein Methods and Protocols - P15
Ngày tải lên :
08/10/2012, 10:25
... hygroscopic and must be thoroughly dried before weighing and making up as standards Standards of mM each of 2-, 3-, and 6-sulfo-Nacetylglucosamine in water may be aliquoted and stored at –20°C, and used ... I., and Nawa, Y (1995) Reserpine-induced sulphomucin production by goblet cells in the jejunum of rats and its significance in the establishment of intestinal helminths Parasite Immunol 17, 581–586 ... phosphate-buffered saline!), as well as sulfate-phosphate-containing detergents (e.g., sodium dodecyl sulfate), chaotropes (e.g., thiourea), and reducing agents (e.g., mercaptoethanol) Ideally, desalting into...
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Glycoprotein Methods and Protocols - P16
Ngày tải lên :
08/10/2012, 10:25
... Clonal Carcinoma Cell Lines, Including LS174T, Caco-2, and A431 Cell Lines (see Notes 8–10) Grow the carcinoma cell lines in tissue culture plastics without collagen coating and 3T3 feeders in DMEM ... FBS and mM of glutamine (Subheading 2., item 5e) Carry out passage as single cells using 0.1% trypsin in 0.1% EDTA (see Notes and 10) 3.5 Apoptotic and Differentiating Cells 3.5.1 Isolation and ... of penicillin and streptomycin, and 50 µg/mL of gentamicin c Digesting solution: The same as the washing medium but containing 5% FBS together with 1.5 mg/mL of collagenase (Worthington type...
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Glycoprotein Methods and Protocols - P17
Ngày tải lên :
08/10/2012, 10:25
... Proteoglycan degrading enzymes and incubation buffers Protease inhibitors may be included in the incubation buffers (see Notes and 2) a Chondroitinase ABC from Proteus vulgaris (Boehringer Mannheim, ... (see Notes and 2) 3.1.1.1 CONTINUOUS LABELING USING GASTROINTESTINAL (GI) CELLS Cells are cultured in standard growth medium as described in Chapter 18 Radioactive precursors (Subheading 2., item ... Labeling of Mucin Precursors and Mature Mucins 3.1.2.1 PULSE/CHASE LABELING USING CELL LINES, PARTICULARLY LS174T, CACO-2, AND A431 Culture cells as described in Chapter 18 Remove the medium and...
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Tìm thêm:
- hệ việt nam nhật bản và sức hấp dẫn của tiếng nhật tại việt nam
- xác định các mục tiêu của chương trình
- xác định các nguyên tắc biên soạn
- khảo sát các chuẩn giảng dạy tiếng nhật từ góc độ lí thuyết và thực tiễn
- khảo sát chương trình đào tạo của các đơn vị đào tạo tại nhật bản
- khảo sát chương trình đào tạo gắn với các giáo trình cụ thể
- xác định thời lượng học về mặt lí thuyết và thực tế
- tiến hành xây dựng chương trình đào tạo dành cho đối tượng không chuyên ngữ tại việt nam
- điều tra đối với đối tượng giảng viên và đối tượng quản lí
- điều tra với đối tượng sinh viên học tiếng nhật không chuyên ngữ1
- khảo sát thực tế giảng dạy tiếng nhật không chuyên ngữ tại việt nam
- khảo sát các chương trình đào tạo theo những bộ giáo trình tiêu biểu
- nội dung cụ thể cho từng kĩ năng ở từng cấp độ
- xác định mức độ đáp ứng về văn hoá và chuyên môn trong ct
- phát huy những thành tựu công nghệ mới nhất được áp dụng vào công tác dạy và học ngoại ngữ
- mở máy động cơ lồng sóc
- mở máy động cơ rôto dây quấn
- các đặc tính của động cơ điện không đồng bộ
- hệ số công suất cosp fi p2
- đặc tuyến hiệu suất h fi p2