Ito 2007 wt LD PRR9 protein

Property Value
BioDare ID 13227547204752
Author Ito S
Institution Nagoya University
License CC_BY

Description

Purpose

PRR9 mRNA and protein were measured under the same conditions. This file has PRR9 protein only.

Comments

This PRR9 protein profile rises more slowly than the FLAG-tagged PRR9 protein in Nakamichi Plant Cell 2010.

Sample preparation

Plants were ground to a fine powder in liquid nitrogen. Crude protein extracts were prepared in 2 vols. of SDS loading buffer (2% SDS, 175 mM Tris–HCl pH 8.0, 0.6 mM EDTA pH 8.0, 3.2% 2-mercaptoethanol, 25% glycerol, 0.008% bromophenol blue). Extracts were immediately boiled for 10 min and the supernatant collected after centrifugation at 6,000×g for 10 min.

Measurement

Protocol

Western blot (TAP-tagged protein)

Literature data from: '' by: .

    These protein samples were separated by 10% SDS–PAGE and transferred onto a PVDF membrane (for each examination, 80 μg of total protein was used). The protein A domains of the TAP-tag were detected with a PAP (peroxidase–anti-peroxidase) antibody (Sigma). Immunoreactive bands were detected by the ECL plus Western blotting system (Amersham Pharmacia Biotech, USA).

Experimental conditions

LD standard

Growth on not entered for 10.0 days (LD standard).

Light

Type Duration (days) Cycle (h) Start Duration Spectrum Source Intensity
diurnal light 10 24 0:00 12:00 white LED 70

Temperature

Type Duration (days) Cycle (h) Base (°C) Warm (°C) Warm start Warm duration
constant temperature 10 24 20 -- -- --

12:12

Growth on not entered for 1.0 days (12:12).

Light

Type Duration (days) Cycle (h) Start Duration Spectrum Source Intensity
diurnal light 1 24 0:00 12:00 white LED 40

Temperature

Type Duration (days) Cycle (h) Base (°C) Warm (°C) Warm start Warm duration
constant temperature 1 24 20 -- -- --

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