analytical methods and skills

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... mammary gland, defined by antibodies to cytokeratins J Cell Sci 75, 17–33 Kiernan, J A (1990) Histological and Histochemical Methods: Theory and Practice, Pergamon, Oxford, UK Andrew, S M and Jasani, ... M and Milton, J D (1986) Lectin Methods and Protocols, Humana, Totowa, NJ Liener, I E., Sharon, N., and Goldstein, I J (1998) The Lectins: Properties, Functions, and Applications in Biology and ... such as Culling’s Handbook of Histopathological and Histochemical Techniques (including museum techniques) (36) or Kiernan’s Histological and Histochemical Methods: Theory and Practice (45) provide...

Ngày tải lên: 08/10/2012, 10:25

... core protein specificity), and lower sensitivity than slot-blot and immunoassay methods Chap- Detection and Quantitation of Mucins 47 ter (2) discusses these assays and they are mentioned here ... greater sample volume, and superior washing) For quantitation and comparison across blots, a standard in the same buffer as samples should be titrated for use as a standard curve, and samples should ... NJ Nagma, K., Thornton, D J., Khan, N., and Sheehan, J K (2000) Separation and identification of mucins and their glycoforms, in Glycoprotein Methods and Protocols: The Mucins (Corfield, T.,...

Ngày tải lên: 08/10/2012, 10:25

... a small gel, use 1.6 g of agarose and 160 mL of buffer, and for a large gel, use 2.8 g of agarose and 280 mL of buffer Leave to cool before pouring (hand hot) and insert well-forming comb (see ... periodic acid-Schiff (PAS) assay and with a variety of lectins and mucin- or carbohydrate-specific antibodies (for a discussion of the relative merits and drawbacks of these analytical tools, see Chapter ... are 1.5 mm thick and cm wide There is a compromise between band broadness and amount of sample to be loaded Larger amounts of sample tend to yield poorer quality data; i.e., the bands become more...

Ngày tải lên: 08/10/2012, 10:25

... Kadler, K E., and Chapman J A (1985) Mica sandwhich technique for preparing macromolecules for rotary shadowing J Ultrastruct Res 91, 66–76 14 Kawahara K and Tanford C (1966) Viscosity and density ... (Fullerton, CA) SW 40Ti swing-out rotor, and finally unloading and analysis of the tube contents Measure out 18 mL of M GuHCl into one chamber of the gradient maker and an equal volume of M GuHCl into ... into 0.5-mL fractions, and these were analyzed for reactivity with antisera for the MUC5AC (᭹) and MUC5B (᭺) mucins The MUC5B mucin is more polydisperse than the MUC5AC mucin and has molecules of...

Ngày tải lên: 08/10/2012, 10:25

... tandem repeat domain and overlapping each other by six amino acids (16), and the other used octamers overlapping each other by seven amino acids (17) Twelve antibodies were analyzed in total and ... information on antibody specificity and permits identification of significant features of an epitope that may contribute to antibody recognition and binding (see Notes 10 and 11) 1.3 Purification of ... must be pure and free from amines Several methods may be used to purify DMF (see Note 16) Piperidine 20% v/v: used for Fmoc deprotection Piperidine should be redistilled before use and made up...

Ngày tải lên: 08/10/2012, 10:25

... lysates and media Nonspecific bands may be present and can be eliminated from consideration using the pSVL vector control lanes for comparison (Fig 2A,B, lanes and 3) The sizes of specific bands ... polymerase buffer, 2.5 mM MgCl2, 400 µM dNTPs, 200 ng of primers and or and 4, ng of template DNA (with only primers and 4), and U of Taq polymerase Perform a PCR program in a suitable thermal ... polymerase, and incubate the reaction at 72˚C to allow extension of the linked templates Add µg each of primers and 6, and continue the PCR with 30 cycles at 94˚C for 30 s, 50˚C for and 72˚C for...

Ngày tải lên: 08/10/2012, 10:25

... was introduced in the 1970s and rapidly received a large application for analyzing the sequence of N-acetyllactosamine- and oligomannosidic-type glycans Originally, and before the development ... appearence of novel methods of ionization such as fast atom bombardment ionization (FAB), electrospray ionization (ESI), and matrixassisted laser desorption ionization (MALDI) Methods 2.1 Nuclear ... method was developed by Vliegenthart and colleagues during the 1970s and essentially applied to the structure determination of N-glycans of the Nacetyllactosamine and oligomannoside type (16) More...

Ngày tải lên: 08/10/2012, 10:25