By Daniele D'Ambrosio, Francesco Sinigaglia
Chemokines and their receptors play a valuable function within the pathogenesis of diverse acute and persistent inflammatory illnesses. This e-book presents sensible protocols for the learn of chemokine receptor biology.
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Extra resources for Cell Migration in Inflammation and Immunity
A) The total ion current (TIC) chromatogram; (B) the sum of the chromatogram tracks for the three major ions of intact synthetic LD78β (ions of 1300, 1560, and 1949 that correspond respectively to six, five, and four times protonated peptide). 1% acetic acid in water/ acetonitrile (50/50) and injected on an ion-trap MS at a flow rate of 5 µL/min. The percent conversion of intact to truncated chemokines is calculated from the relative intensity of the signals on the MS (see Fig. 6) (see Note 9).
FACScalibur®; Becton Dickinson, Mountain View, CA). 16. , CellQuest® software; Becton Dickinson). 2. Measurement of the Effect of GFP-Tagged Proteinsin the Content of F-Actin 1. , Palo Alto, CA], expressing GFP alone or fused to wild-type Rac1, V12Rac1, N17Rac1, wild-type Cdc42, V12Cdc42, and N17Cdc42). 2. Standard molecular-biology reagents and procedures for the generation of transfection-grade plasmid DNA. 3. Optimem® medium (Life Technologies, Paisley, UK). 4. , GenePulser; Bio-Rad, Hercules, CA).
With both physical parameters in a linear scale, the beads will be located in the upper side of the acquisition dot plot; with both parameters in a logarithmic scale, beads will be visible on the upper right corner of the acquisition dot plot (see Fig. 1). 8. Draw a region around cells (region 1, R1) and a region around Polybeads (R2) (see Fig. 1). 9. Set acquisition parameters to acquire 3 × 104 events in R2 and to store events of R1 plus R2. 10. Acquire samples with a sample flow rate of 200 µL/min.