Kidney Research: Experimental Protocols by Tim D. Hewitson, Edward R. Smith, Stephen G. Holt (eds.)

By Tim D. Hewitson, Edward R. Smith, Stephen G. Holt (eds.)

This moment variation expands upon the former quantity with extra emphasis on contemporary innovation in uncomplicated renal examine and has a extra holistic strategy on linked problems resembling problems linked to peritoneal dialysis, ischemic acute kidney damage, the sympathetic fearful process, and vascular calcification. The ebook is split into 5 elements: half I offers a couple of in vitro, in vivo, and ex vivo types of kidney illness and linked problems; half II seems to be at fresh advances in imaging ideas; half III covers contemporary advancements in learning metabolism in renal ischemia and reperfusion; half IV addresses examine and size of vascular calcification; and half V explores analytical suggestions which are either topical and of common relevance to the research of experimental renal affliction. Written within the hugely profitable Methods in Molecular Biology series structure, chapters comprise introductions to their respective issues, lists of the mandatory fabrics and reagents, step by step, without difficulty reproducible laboratory protocols, and pointers on troubleshooting and fending off identified pitfalls.

Authoritative and state-of-the-art, Kidney examine: Experimental Protocols, moment Edition is a important selection of protocols helpful to new and skilled researchers who're attracted to the sphere of Nephrology.

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8. , Dako Pen; Dako, Glostrup, Denmark). 9. Humidified chamber (a sealed box with paper tissue at the bottom and soaked with ddH2O to maintain humidity). 2 Immuno fluorescence 1. Antibodies (Table 1). 2. Fixatives: methanol, 4 % PFA. 3. Permeabilizing solution: Triton X-100. 4. Blocking buffer: 1× PBS containing 3 % BSA (w/v) and 10 % goat serum (v/v). Table 1 Cell characterization Antibody IgG source Dilution Application Company α-SMA Mouse 1:200 WB/IF Sigma Cytokeratin 18 Mouse 1:100 WB/IF Sigma Vimentin Mouse 1:100 WB/IF Sigma Alexa 488 Goat 1:1000 IF Invitrogen WB, western blotting; IF, immunofluorescence Isolation and Propagation of Rat Peritoneal Mesothelial Cells 29 5.

Perit Dial Int 25:S14–S18 2. Nishimura M, Ito Y, Mizuno M et al (2008) Mineralocorticoid receptor blockade ameliorates peritoneal fibrosis in new rat peritonitis model. Am J Physiol Renal Physiol 294: F1084–F1093 3. Sawai A, Ito Y, Mizuno M et al (2011) Peritoneal macrophage infiltration is correlated with baseline peritoneal solute transport rate in peritoneal dialysis patients. Nephrol Dial Transplant 26:2322–2332 4. Mizutani M, Ito Y, Mizuno M et al (2009) Connective tissue growth factor (CTGF/ CCN2) is increased in peritoneal dialysis patients with high peritoneal solute transport rate.

The dissociation of cells from the surface should be checked by direct visualization using a light microscope. When lifting cell monolayers from subsequent subcultures, the cells should be left to incubate with the EDTA/trypsin solution for a minimum of 5 min and no longer than 10 min. Prolonged incubation with EDTA/trypsin solution may cause cellular damage. 5. When initial primary human renal fibroblasts are 100 % confluent, the cell monolayer can be passaged using the trypsinization method described.

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