GLYCOPROTEIN METHODS PROTOCOLS - BIOTECHNOLOGY 048-9-045-055.PDF

Tìm thấy 10,000 tài liệu liên quan tới tiêu đề "Glycoprotein methods protocols - biotechnology 048-9-045-055.pdf":

GLYCOPROTEIN METHODS PROTOCOLS - BIOTECHNOLOGY 048-9-261-271

GLYCOPROTEIN METHODS PROTOCOLS - BIOTECHNOLOGY 048-9-261-271

3. Methods
3.1. Cell Culture in the Presence of GalNAc α -O-benzyl
3.1.1. Conditions of Use of GalNAc α -O-benzyl
GalNAc α - O -benzyl is directly dissolved in the culture medium for 2 h at room temperature with continuous stirring. Then, the medium is sterilized by filtration. GalNAc α[r]

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GLYCOPROTEIN METHODS PROTOCOLS - BIOTECHNOLOGY 048-9-323-336

GLYCOPROTEIN METHODS PROTOCOLS - BIOTECHNOLOGY 048-9-323-336


Currently, the two probes of choice for ISH are riboprobes of usually 100–300 bp (RNA transcribed from cDNA probes) or oligonucleotide probes of 18–100 bp (which match the cDNA sequence), the latter being less sensitive. Because of the enhanced signal obtained with TR probes, straightforward simpl[r]

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GLYCOPROTEIN METHODS PROTOCOLS - BIOTECHNOLOGY 048-9-385-392

GLYCOPROTEIN METHODS PROTOCOLS - BIOTECHNOLOGY 048-9-385-392

8. The use of phosphate buffers should be avoided if possible because phosphate may inhibit sulfatase activities ( see Chapter 34). and thus affect overall mucinase activity.
9. Blank incubations can be prepared using enzyme extract that has been heat treated for 5 min at 95 ° C and centri[r]

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GLYCOPROTEIN METHODS PROTOCOLS - BIOTECHNOLOGY 048-9-369-381

GLYCOPROTEIN METHODS PROTOCOLS - BIOTECHNOLOGY 048-9-369-381

12. Rentrop, M., Knapp, B., Winter, H., and Schweizer, J. (1986) Aminoalkylsilane-treated slides as support for in situ hybridization of keratin cDNAs to frozen tissue sections under varying fixation and pretreatment condition. Histochem. J. 18, 271–276.
13. Briand, J. P., Muller, S., and Van Re[r]

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GLYCOPROTEIN METHODS PROTOCOLS - BIOTECHNOLOGY 048-9-337-350

GLYCOPROTEIN METHODS PROTOCOLS - BIOTECHNOLOGY 048-9-337-350

and Corstedt, I. (1999) Studies on the insoluable glycoprotein complex from human colon: identification of reduction-insensitive MUC2 oligomers and C-terminal cleavage. J. Biol. Chem. 274, 15,828–15,836.
28. Gum, J. R., Hicks, J. W., Toribara, N. W., Siddiki, B., and Kim, Y. S. (1994) Molec[r]

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GLYCOPROTEIN METHODS PROTOCOLS - BIOTECHNOLOGY 048-9-403-416

GLYCOPROTEIN METHODS PROTOCOLS - BIOTECHNOLOGY 048-9-403-416

sodium acetate, pH 4.0); coffee bean α -galactosidase (200 m M Tris-HCl, pH 6.5); bovine testes (100 m M citrate phosphate, pH 4.0).
11. In addition to the colorimetric assays given in this chapter, additional sensitivity can be attained by the use of fluorimetric assays with 4-methyl umbellifer[r]

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GLYCOPROTEIN METHODS PROTOCOLS - BIOTECHNOLOGY 048-9-313-321

GLYCOPROTEIN METHODS PROTOCOLS - BIOTECHNOLOGY 048-9-313-321

4. Ethidium bromide is not added to DNA before electrophoresis because it is thought to modify the migration of DNA.
Acknowledgments
The methods described here were developed with the help of Veronique Guyonnet- Duperat (20) and Pascal Pigny (21) . The authors thank Dr. Alexander S. Hill[r]

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Glycoprotein methods protocols - biotechnology 048-9-305-312.pdf

GLYCOPROTEIN METHODS PROTOCOLS - BIOTECHNOLOGY 048-9-305-312.PDF

1. Introduction
Northern blot analysis has historically been one of the most common methods used to provide information on the number, length, and relative abundance of mRNAs expressed by a single gene. This technique also generates a record of the total mRNA content expressed by a cell cul[r]

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GLYCOPROTEIN METHODS PROTOCOLS - BIOTECHNOLOGY 048-9-297-303

GLYCOPROTEIN METHODS PROTOCOLS - BIOTECHNOLOGY 048-9-297-303

3. Methods
3.1. Preparation of mRNA
1. Total RNA is isolated from cultured cells or human mucosae as described in Chapter 25. In spite of the fact that the purification yield of poly A + mucin fraction is very poor, the purification step of polyA + RNA from total RNA cannot be omitted pri[r]

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GLYCOPROTEIN METHODS PROTOCOLS - BIOTECHNOLOGY 048-9-273-293

GLYCOPROTEIN METHODS PROTOCOLS - BIOTECHNOLOGY 048-9-273-293

TRANG 12 BROCKHAUSEN TABLE 1 MUCIN GLYCOSYLTRANSFERASES, THEIR SUBSTRATES AND PRODUCTS, AND HPLC CONDITIONS FOR PRODUCT ISOLATION HPLC Path Enzyme Substrate Product column % AN a Polypep[r]

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GLYCOPROTEIN METHODS PROTOCOLS - BIOTECHNOLOGY 048-9-001-013

GLYCOPROTEIN METHODS PROTOCOLS - BIOTECHNOLOGY 048-9-001-013

5. Monitor the eluate on-line with a UV monitor and collect fractions using a fraction col- lector and subject to the appropriate carbohydrate and antibody analyses.
3.6. Analysis of Mucins
Methods for the detection and analysis of mucins are dealt with in other chapters in this volume. H[r]

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GLYCOPROTEIN METHODS PROTOCOLS - BIOTECHNOLOGY 048-9-393-401

GLYCOPROTEIN METHODS PROTOCOLS - BIOTECHNOLOGY 048-9-393-401


8. It is essential to have isotonic conditions, i.e., 300 mosmolar to shield the negative charge on the mucins otherwise anomalously high viscosities will result.
9. It is vital to include controls without enzyme in the mucolysis experiments. Crude mucus samples may contain endogenous pro[r]

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Glycoprotein methods protocols - biotechnology 048-9-471-486.pdf

GLYCOPROTEIN METHODS PROTOCOLS - BIOTECHNOLOGY 048-9-471-486.PDF


MUC1/neo-positive population of cells sorted by flow cytometry died within 2 wk with- out proliferation. Cloning of MUC1/neo-positive cells by limiting dilution in a 96-well plate resulted in progressive death of these cells, whereas neo-positive only cells expanded. We do not have a good explanat[r]

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GLYCOPROTEIN METHODS PROTOCOLS BIOTECHNOLOGY 048 9 495 500 PDF

GLYCOPROTEIN METHODS PROTOCOLS BIOTECHNOLOGY 048 9 495 500 PDF

8. Horseradish peroxidase-labeled rabbit antihuman IgG and IgM, both from Dako A/S, Glostrup, Denmark. Dilute the conjugate 1:10,000 in phosphate/salt buffer containing 0.5% BSA. 9. Substrate solution containing 0.06 mg/mL of tetramethylbenzidine (TMB) diluted in 0.1 M
sodium acetate/citric[r]

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GLYCOPROTEIN METHODS PROTOCOLS - BIOTECHNOLOGY 048-9-487-493

GLYCOPROTEIN METHODS PROTOCOLS - BIOTECHNOLOGY 048-9-487-493

3. Transfer the marrow into a 20-mL universal. Allow any fragments of bone or muscle to fall to the bottom of the tube under gravity and the transfer the supernatant to a fresh universal. 4. Count cells and resuspend at 3.33 × 10 5 /mL in IMDM ( see Subheading 2., item 1e ) plus 5%FCS and mouse GM-[r]

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GLYCOPROTEIN METHODS PROTOCOLS BIOTECHNOLOGY 048 9 463 470 PDF

GLYCOPROTEIN METHODS PROTOCOLS BIOTECHNOLOGY 048 9 463 470 PDF

In addition to the MHC-unrestricted CTLs, some MUC1-specific MHC-restricted CTLs have also been identified. Patients who are HLA-A11 and HLA-A3 have CTLs that recognize a nine amino acid peptide from the tandem repeat region bound to these alleles (6) . Another peptide has been reported that binds H[r]

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GLYCOPROTEIN METHODS PROTOCOLS BIOTECHNOLOGY 048 9 455 462 PDF

GLYCOPROTEIN METHODS PROTOCOLS BIOTECHNOLOGY 048 9 455 462 PDF

T.L đi, đứng, bị, lẫy theo cữ và cịn hơi nhanh hơn so với các trẻ khác. Được 2 tháng tuổi cháu đã biết lẫy, 9 tháng tuổi cháu đã biết đi tốt và biết nĩi khoảng 5 từ. Gọi “T.L ơi” cháu biết “ dạ” rất rõ ràng. Hỏi cháu cĩ ăn đồ gì đĩ khơng cháu biết nĩi “ khơng ” hoặc “ cĩ”. Khi xem phim[r]

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GLYCOPROTEIN METHODS PROTOCOLS BIOTECHNOLOGY 048 9 439 452 PDF

GLYCOPROTEIN METHODS PROTOCOLS BIOTECHNOLOGY 048 9 439 452 PDF

Methods outlined for the isolation of bacteria from the rectal mucosa are essentially destructive, and primarily provide details of the types and numbers of different species that take part in this process. They do not contribute information concerning the mul- ticellular organization of biof[r]

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GLYCOPROTEIN METHODS PROTOCOLS BIOTECHNOLOGY 048 9 429 437 PDF

GLYCOPROTEIN METHODS PROTOCOLS BIOTECHNOLOGY 048 9 429 437 PDF

2. Wash wells 24 times with PBS.
3. Block for 2 h at room temperature (or overnight at 4 ° C) by adding 200 µ L of 1% BSA/ PBS to each well.
4. After blocking, wash wells twice with PBS/Tween and incubate with 100 µ L of 5 µ g/mL biotinylated wheat germ agglutinin (WGA) for 45 min at room tem[r]

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GLYCOPROTEIN METHODS PROTOCOLS - BIOTECHNOLOGY 048-9-417-426

GLYCOPROTEIN METHODS PROTOCOLS - BIOTECHNOLOGY 048-9-417-426

THE HISTOCHEMICAL DESCRIPTION OF MUCIN AS NEUTRAL, SIALO-, AND SULFOMUCIN DEPENDS ON WHETHER THE MIXTURE OF OLIGOSACCHARIDES IN THE MOLECULE, AND INDEED THE MIXTURE OF MUCIN MOLECULES PR[r]

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