By Andrew P. Cope
Here's a compendium of information pertinent to the tools and protocols that experience contributed to either fresh advances in molecular medication typically in addition to to molecular foundation of rheumatic ailment specifically. This two-volume paintings collects the contributions of leaders within the box who hide such interesting and leading edge issues as imaging and immunohistochemistry, research of cartilage and bone catabolism, immunobiology, and telephone trafficking.
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Additional resources for Arthritis Research: Volume 2: Methods and Protocols (Methods in Molecular Medicine)
2. Tissues are cut into 6 µm sections to get a cell monolayer. 3. Sections are thaw-mounted onto adhesive slides and air-dried for 2 h. 4. Fix the tissue sections in ice cold acetone for 10 min and air dry 5. ) (see Note 1). Alternatively, the frozen section can be stored for about 1 mo at –70°C. 6. To prevent rehydration of tissue sections during storage, put CaCl2 wrapped in tissue paper into the box. 2. Immunohistochemistry Immunohistochemistry permits the identification of cell types infiltrating the synovial tissue, and the analysis of their organization in situ (see Note 2).
2. Incubate with Cy3-conjugated streptavidin diluted in TBS. From this step and onwards light should be eluded to preserve the fluorochromes (see Note 9). 3. Rinse sections in TBS; once briefly and once for 5 min. 4. Incubate with FITC-conjugated anti-IgG/IgA/IgM antibodies. 5. Rinse sections in TBS; once briefly and once for 5 min. 6. Mount in anti-fading medium. 5. Analysis of Slides Computer assisted image analysis is suitable for determining degree of overlap when double staining of antigen specificity and isoform is performed.
664120). 19. Amicon Ultra-15 centrifugal filter unit 30,000 MW cutoff (Millipore; cat. no. UFC903096). 3 Peptide Loading and Chromophore-Labeled Tetramer Formation 1. Class II binding peptide at 50 mg/mL in dimethyl sulfoxide (DMSO) (approx 15 mer peptide). 2. 9). 3. Pefabloc SC: 100 mM in H2O (Boehringer Mannheim; cat. no. 1585-916). 42 Gebe and Kwok 4. OG. 5. Phycoerythrin-labeled streptavidin (BioSource International; cat. no. SNN 1007). 3. 1. 1. pRmHa-3 Construct The copper-inducible metalothionine promoter vector pRmHa-3 is shown in Fig.
Arthritis Research: Volume 2: Methods and Protocols (Methods in Molecular Medicine) by Andrew P. Cope