By Alexandre R. Gingras, Feng Ye, Mark H. Ginsberg (auth.), Amanda S. Coutts (eds.)
Cellular adhesion is a basic approach that impacts a variety of organic actions equivalent to morphogenesis, telephone motility and department, in addition to signalling. furthermore, adhesion is a method vital not just in basic body structure and improvement, but in addition in sickness states similar to tumourigenesis, heart problems, irritation and an infection. There are a plethora of proteins all in favour of adhesion-related occasions with a tremendous variety in functionality. for this reason, a wide selection of innovations exist to review adhesion comparable proteins and methods. In Adhesion Protein Protocols, 3rd Edition, chapters conceal suggestions to realize perception into the complicated and incompletely understood methods which are enthusiastic about mobile adhesion. Written within the winning Methods in Molecular Biology sequence layout, chapters comprise introductions to their respective themes, lists of the required fabrics and reagents, step by step, quite simply reproducible protocols, and notes on troubleshooting and heading off recognized pitfalls.
Authoritative and simply available, Adhesion Protein Protocols, 3rd Edition could be necessary for either these new to the sector of adhesion protein learn in addition to the more matured scientist.
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Extra info for Adhesion Protein Protocols
Adams JC (2002) Regulation of protrusive and contractile cell-matrix contacts. J Cell Sci 115:257–265 2. Lauffenburger DA, Wells A (2001) Getting a grip: new insights for cell adhesion and traction. Nat Cell Biol 3:110–112 3. Helenius J, Heisenberg CP, Gaub H et al (2008) Single-cell force spectroscopy. J Cell Sci 121:1785–1791 4. Benoit M, Gabriel D, Gerisch G et al (2000) Discrete interactions in cell adhesion measured by single-molecule force spectroscopy. Nat Cell Biol 2:313–317 5. Taubenberger A, Cisneros DA, Friedrichs J et al (2007) Revealing early steps of alpha2beta1 integrin-mediated adhesion to collagen type I by using single-cell force spectroscopy.
Quantifying Cell Adhesion by SCFS 31 3. Retract the cantilever ≈80 μm to exclude negative impacts of long-range interactions from the surface on the subsequent cantilever spring constant calibration. 4. Determine the cantilever spring constant using the routines incorporated into the AFM control software (see Note 34). 3 Attaching a Cell to Cantilever 1. Lift the AFM head and inject ≈1 × 103 cells into the PetriDishHeater™ right on top of the BSA-coated spot. 2. Immediately place the AFM head on the CellHesion™ stage and manually position the apex of the cantilever above a suitable cell (Fig.
Penicillin. 3. Streptomycin. 4. PBS. 5. 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES). 6. HeLa cells (ATCC). 25 Quantifying Cell Adhesion by SCFS 7. Fetal bovine serum (FBS). 8. DMEM (containing GlutaMAX™, 4,500 mg/L D-glucose, sodium pyruvate, NaHCO3): Mix DMEM with 10 % FBS, 100 U/mL penicillin and 100 μg/mL streptomycin. 22 μm filter (see Note 7). 9. CO2-independent AFM medium: Dissolve DMEM powder (containing glutamine, 4,500 mg/L D-glucose, sodium pyruvate, no NaHCO3) in the appropriate amount of ultrapure water.