MBSA

Methylated bovine serum albumin (MBSA) MBSA 

Immunogen is complexed by the addition of an equal amount (w/w) of MBSA and the precipitate is resuspended to a concentration of 0.25 mg/ml before it is emulsified with Freund s adjuvant. 

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MARCKS Myristolated, alanine-rich C kinase substrate specific protein kinase C substrate MBP Major basic protein MBSA Methylated bovine serum albumin... 

FAB-MS is a renewed old technique, first described in 1966 as molecular beam for solid analysis (MBSA) [73], and later (1981) further extended with a liquid matrix [72,74], The FAB experiment is closely related to... 

Figure 74. Percent Moisture-Equivalent and Water-holding Capacity in Sand-silt Mixtures. (Yuma-mbsa Sandy Soil.) Numbers Refer to Analyses Given in Table 55.

Methylated bovine serum albumin (mBSA) (from Sigma) Dissolve in water (1 mg/mL) and store at-20°C. 

For every 1 mL of this solution, add 15 pL of mBSA solution, keeping the soludon agitated during the addidon. The soludon immediately becomes cloudy as the result of the formadon of an electrostadc complex between the mBSA and DNA see Note 8). 

Three or four days before carrying out a spleen cell fusion give the chosen animal an iv injection (0.5 mL) of a suspension of DNA/mBSA complex prepared as in Step 2. 

Peyer s Patch Immunization The DNA solution should be at a concentration of about 1 mg/mL see Note 11). Allow about 80 pL of DNA solution per rat per immunization. Add mBSA solution, such that the weights of mBSA and DNA are equal. Then add 1.1 vol of adjuvant and emulsify as described in Step 3 of the previous section, taking care not to make the emulsion too viscous, as it must be injected via a very fine needle. 

Immunization with denatured DNA-MBSA gives rise to antibodies, often mainly of the IgM class, that react with single-stranded DNA but not with native DNA. The largest antigenic determinant for such antibodies is about the size of a pentanucleotide. When DNA with an unusual base, such as the glucosylated hydroxymethylcytosine of T-even phage, is used, the specificity is directed largely to the modified base and much more IgG may be produced.Similarly, when UV-irradiated DNA or photooxidized DNA is used, the modified bases of the lesions provide the major specificity determinants. - ... 

Immunization with homopolynucleotide-MBSA complexes gives rise to antibodies that are specific for an oligonucleotide segment of the polymer. They probably recognize a number of bases in sequence, perhaps in a stacked array. Poly(I), poly(C), and poly(A) each induce specific antibodies that show little cross-reaction with the other homopolynucleotides, and slight cross-reaction with denatured DNA. ... 

Procedures for preparation of the immunizing antigen, injection schedules, and assays for antibody formation are similar to those used for helical polynucleotide-MBSA antigens and are discussed in detail in the following section and in Vol. 12B [174] of this series. 

For preparation of the aggregated polynucleotide-MBSA complex, it is convenient to use the polynucleotide at a concentration of about 200-500 /ig/ml. A stock solution of 10 mg of MBSA per milliliter in water is prepared the powdered or lyophilized MBSA does not dissolve readily in saline. An amount of MBSA equal to the total weight of the polynucleotide is added to the annealed polymer. A white precipitate should be visible after the MBSA is added. It is an easily handled fine suspension with most polymers, but fibrous strands may be formed with samples of very high molecular weight. Preliminary mechanical homogenization or soni-cation usually reduces such polymers to a size that will form a manageable suspension with the MBSA. 

The polynucleotide-MBSA suspension is then emulsified with an equal volume of Freund s adjuvant until a stable thick white emulsion is formed. Complete adjuvant is used for the first immunization, and incomplete adjuvant for subsequent injections. 

A primary dose of 50-200 fig of polynucleotide, in the MBSA complex and emulsified with complete adjuvant, is injected into each rabbit at several intradermal sites along the back and subcutaneously a total volume of 1 ml per rabbit is convenient. Similar doses, but with incomplete adjuvant, are given intradermally and subcutaneously on days 14 and 21, and the animals are bled 5-7 days later. They can be bled at weekly intervals, and additional booster immunizations can be given if antibody levels should fall. During early courses of immunization, they may peak and fall rapidly. ... 

In contrast to single-stranded DNA, native DNA adsorbs poorly to polystyrene (Engvall, 1976). This problem can be overcome by treating the polystyrene with a 1% solution of aqueous protamine sulfate for 90 min (Klotz, 1982), followed by three washes with distilled water and drying. Other polycationic substances, such as MBSA (Section 5.2.3.3, Ferrua et al., 1983) or poly-L-lysine (type VII B Sigma), may also be used (Leipold et al., 1983). Poly-L-lysine is suspended in water at a concentration of 1 mg/ml (which may... 

Fast atom bombardment mass spectrometry (FAB-MS) is a renewed old technique that is most often thought of in terms of a relatively novel mass spectral technique. In fact, as early as in 1966, Devienne and co-workers presented data on a technique that they called molecular beam for sohd analysis (MBSA) [1]. The idea was further developed by the same group but was ignored by the rest of the scientific community until the announcement by Barber et al. [2] and by Surman and Vickerman [3] of the discovery of FAB stimulated efforts throughout the world as well as a number of international symposia and workshops devoted in whole, or in part to the understanding of FAB and its apphcations. 

Exogenous maltosyl bovine serum albumine at two concentrations. MBSA (Sigma Chemical lot 98F8120) is prepared from bovine albumin and maltose coupled via reductive amination.8 One mole of albumin contains 14 moles of disaccharide linked to lysine residues. The sugar content represents 5% of the molecule. The MBSA molecular weight and p/ are 71 kDa and 4, respectively. It was used without further purification. 

The foam parameters of the YGP solutions have the same order of magnitude as the base wine ones although the parameters of the more concentrated solutions are higher. The MBSA solution parameters are drastically higher. They were not measurable with our device for the higher MBSA concentration besides, some gel-like liquid remained stuck on the wall of the test tube. 

It is remarkable that the 1993 and 1994 base wines compare so well within the experimental errors, at the local scale with same kinetics of film drainage and same aggregates, and at the global scale with same order of magnitude of the three foam parameters. It is amazing that the foam expansion E, the foam lifetimes Ln and L(2, and the foamability X all have smaller values with the 1990 wine than with the two others. Not less remarkable is the comparison between the 1990 and the dilute YGP model solution, the main difference between the wine and the model solution being the existence of residual foams with wines which do not exist in the case of the model solution. And finally the behaviours of the films and foams of the MBSA solutions are completely different from the wine and YGP solutions ones. 

Sulfatide Lecithin -1- cholesterol + MBSA None Ab RB Zalc et al. (1977)... 

T riphosphoinositide Lecithin+cholesterol -h MBSA None Ab RB Greenberg et al. (1979)... 

MBSA=methylated bovine serum albumin CFA=complete Freund s adjuvant DH = delayed hypersensititivy Ab=antibody response RB=rabbits GP=guinea pigs H = humans IFA=incomplete Freund s adjuvant... 

The first step is the preparation of the emulsion containing the adjuvant and the antigen. For each mouse, 500 pg of mBSA must be dissolved in a final volume of 100 pL (50 pL of CFA+ 50 pL of PBS). There are significant losses during the preparation and injection and this must be taken into consideration when deciding the amount of material to be prepared. 

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