Recipe

Motility Solution

10–1000 pm bifunctional rhodamine-calmodulin (BR-CaM)-labeled myosin V, prepared as in Fluorescent Labeling of Myosin V for Polarized Total Internal Reflection Fluorescence Microscopy (polTIRFM) Motility Assays (Beausang et al. 2012)

2.5 mg/mL bovine serum albumin (BSA; Sigma-Aldrich A0281)

100 mm DTT (prepare 1 m DTT stock in M5 buffer daily)

100 µg/mL wild-type calmodulin (WT-CaM)

1–200 µm ATP

Prepare daily in M5 buffer.

REFERENCE

  1. ↵
    1. Beausang JF,
    2. Sun Y,
    3. Quinlan ME,
    4. Forkey JN,
    5. Goldman YE
    Beausang JF, Sun Y, Quinlan ME, Forkey JN, Goldman YE. 2012. Fluorescent labeling of myosin V for polarized total internal reflection fluorescence microscopy (polTIRFM) motility assays. Cold Spring Harb Protoc doi:10.1101/pdb.prot069369.
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  1. doi:10.1101/pdb.rec070193 Cold Spring Harb Protoc 2012: pdb.rec070193- © 2012 Cold Spring Harbor Laboratory Press
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