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

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]

The methods for electrophoresis of RNA have been described in many books. The protocol below represents a modified version of the standard technique described in ref. 6 , and only the modifications introduced to fractionate mucin RNAs are described in detail. 1. Prepare the RNA samples: The[r]

themselves may sometimes complicate the picture. The detailed protocols are given in
Subheading 3. Full protocols for MUC7 are not given, but the appropriate literature is cited ( see Note 8).
Although at present the only way of analyzing this variation is by Southern blotting,[r]

9. Gipson, I. K. and Inatomi, T. (1999) Cellular origin of mucins of the ocular surface. in
Advances in Experimental Medicine and Biology (Sullivan, D. A., ed.), Plenum, New York, pp. 221–228.
10. Gipson, I. K., Ho, S. B., Spurr-Michaud, S. J., Tisdale, A. S., Zhan, Q., Torlakovic, E.,[r]

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]

1. Introduction
One of the interesting technical aspects of working with mucins is that it is rela- tively easy to make antibodies to different mucin glycoproteins—mainly because the repeat sequences in the variable numbers of tandem repeat (VNTR) region are highly immunogenic. Indeed, all the muc[r]

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]

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]

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]

In this Chapter, we focus on the identification of each of the known MUC-type mucin precursors by immunoprecipitation using antipeptide antibodies. Moreover, a number of biochemical and cell biological assays will be described which establish the presence in the RER of each alleged MUC-type mucin pr[r]

9. In this procedure, the antibodies present in one sample (particularly the IgG-fraction) will end up in one lane of the gels, which are used to analyze the samples. As a result, the maximal amount of antibody that can be added to one homogenate or medium sample is determined by the amount[r]

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]

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]

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]

An alternative to the radioactive methods is the use of biotinylated mucin sub- strates tagged through either the peptide or carbohydrate moieties of the molecule
(6 , 7) . Such assays have been described for protein substrates and adapted for use with microtiter plates (8) . They require s[r]

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]

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]

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]

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]

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]