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Membrane strategies have broad commercial ap This instruction manual reports the printed litera plications masking many present and rising ture, offers an in-depth description of com makes use of within the chemical, petrochemical, petroleum, mercialized membrane procedures, and provides a state of the art evaluation of latest membrane seasoned environmental, water remedy, pharmaceutic al, scientific, foodstuff, dairy, beverage, paper, tex cess strategies less than improvement.
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5 mmol) of 6-chloropurine ribonucleoside in 21 ml of ethanol while stirring. 2. 5 g (21 mmol)] and stir under a reflux condenser for 12 hr. 3. Remove solvent under reduced pressure in Buchi Rotovapor. 4. Add 150 ml of ethanol and place at 4◦ C for 5 hr. 5. Filter the crystals and wash with cold ethanol to obtain N6 phenethyl (or additional alkyl) adenosine. Synthesize N6 phenethyl ADP 6. 76 g) in 5 ml of triethethylphosphate (TEP). 7. Cool to 0◦ C in an ice/NaCl bath while stirring. 8. 6 g (4 mmol) of POCl3 dropwise and continue to stir at 0◦ C for 2 hr.
PREPARATION OF A THIOPHOSPHORYLATED POSITIVE CONTROL PEPTIDE AND PROTEIN The two control samples, a thiophosphorylated peptide and protein, can be prepared from easily purchased materials. A kinase reaction is used to make the thiophosphorylated substrates. MALDI-MS analysis can be used to confirm whether the CREB peptide has been thiophosphorylated. For the protein reaction, after labeling, the thiophosphorylated myelin basic protein (MBP) can be run on a denaturing polyacrylamide gel followed by an immunoblot using 51-8 (Support Protocol 1).
A-3059) should be included in the buffer to avoid binding of these antibody populations with albumin conjugates on the array surface. 12. Prepare a reference sample that serves as a positive control and will be used later in data analysis. Glycan Microarrays a. Pooled serum from multiple donors can provide a reference when analyzing serum samples. To minimize freeze-thaw cycles for the reference sample, divide a single 44 Volume 2 Current Protocols in Chemical Biology Table 2 Recommended Buffers and Starting Dilutions of Samples for Incubation on Glycan Microarray Sample type Incubation buffera Concentration Lectin 1% BSA in PBST 1-50 μg/ml Room temperature for 2 hr Monoclonal antibody 3% BSA in PBST 1-50 μg/ml 37◦ C while gently shaken for 2-4 hr Serum antibodies 3% BSA in PBST 1:50 to 1:200 Conditions 37◦ C while gently shaken for 4 hr a PBST: PBST: see recipe for PBST array wash buffer in Reagents and Solutions.