Protocol detail

Immunodetection of Epithelial Cytoskeletal Proteins

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Myo1a and Myo1e, 1 μg/ml [Skowron et al. 1998 (link)], Myo1d and sucrase isomaltase, 1.5μg/ml (Santa Cruz Biotechnology, Inc., Dallas, TX), Myo1c and alkaline phosphatase, 1μg/ml (Sigma, St. Louis, MO), villin, 1μg/ml (AMAC, Westbrook, ME), espin, 1μg/ml (Gift from J.R. Bartles, Northwestern University), fimbrin, 1μg/ml [Shibayama et al. 1987 (link)], Myo5a and Myo6, 1μg/ml [Heintzelman et al. 1994 (link)], Myo1a and Myo1e, 1 μg/ml [Skowron et al. 1998 (link)], ezrin, phosphoezrin, and clathrin, 1:1000 (Cell Signaling Technology, Danvers, MA), cytokeratin 8, 1:500 (TROMA-1, DSHB, Iowa City, IA), adaptin beta, 1:500 (BD Biosciences, Franklin Lakes, NJ), NHE3, 1μg/ml (Alpha Diagnostic, San Antonio, TX), NaPi2b 1 μg/ml [Giral et al. 2009 (link)], spectrin, 1:1000 dilution of serum (gift from J. Morrow, Yale School of Medicine). CFTR, AME 4991 CFTR, 1:1000 [Kravtsov et al. 2012 (link)]

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Hegan P.S., Kravtsov D.V., Caputo C., Egan M.E., Ameen N.A, & Mooseker M.S. (2015). Restoration of cytoskeletal and membrane tethering defects but not defects in membrane trafficking in the intestinal brush border of mice lacking both myosin Ia and myosin VI. Cytoskeleton (Hoboken, N.J.), 72(9), 455-476.

Publication 2015

ezrin phosphoezrin clathrin

Alkaline phosphatase Cftr Clathrin Cytokeratin 8 Diagnostic Dilution Ezrin Fimbrin Myo5a Napi2b Serum Spectrin Sucrase isomaltase Villin

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Variable analysis

independent variables

Myo1a and Myo1e
Myo1d and sucrase isomaltase
Myo1c and alkaline phosphatase
Villin
Espin
Fimbrin
Myo5a and Myo6
Myo1a and Myo1e
Ezrin, phosphoezrin, and clathrin
Cytokeratin 8
Adaptin beta
NaPi2b
Spectrin
CFTR, AME 4991 CFTR

dependent variables

Not explicitly mentioned

control variables

Concentrations of proteins: 1 μg/ml, 1.5 μg/ml, 1:1000 dilution

positive controls

None specified

negative controls

None specified

Annotations

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