Arabidopsis CAND1, an Unmodified CUL1-Interacting Protein, Is Involved in Multiple Developmental Pathways Controlled by Ubiquitin/Proteasome-Mediated Protein Degradation

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Arabidopsis CAND1, an Unmodified CUL1-Interacting Protein, Is Involved in Multiple Developmental Pathways Controlled by Ubiquitin/Proteasome-Mediated Protein Degradation

Publisher
American Society of Plant Biologists
Publication Date
Jul 01, 2004
Source
PMC
Keywords
Disciplines
  • Biology
License
Unknown

Abstract

STANDARD OPERATING PROCEDURE http://proteomics.cancer.gov M-101.1 Page 1 of 3 STANDARD OPERATING PROCEDURE Title: Protein concentration measurements using Bio-Rad protein assay kit SOP#: M-101 Version #: 1 Author: A. Stephen / G. Whiteley Date Approved: Feb. 6, 2009 Date Modified: 1. PURPOSE This procedure is to be used for the determination of the protein concentration. 2. SCOPE This document describes the procedure for measuring protein concentration of a sample. 3. RESPONSIBILITIES It is the responsibility of the person(s) performing this test to be familiar with lab safety procedures and to have basic laboratory skills. The interpretation of results must be done by a person trained in the procedure and familiar with such interpretation. 4. EQUIPMENT  Perkin Elmer Lambda20 spectrophotometer 5. MATERIALS  Polystyrene semi-microcuvettes; Sarstedt Cat. # 67.742  Parafilm 6. REAGENTS  Protein to be tested  BioRad Protein Assay concentrated dye reagent; BioRad Cat. #500- 0006  Bovine Serum Albumin (BSA) protein standard; Bio-Rad Cat. #500- 0007 http://proteomics.cancer.gov M-101.1 Page 2 of 3 7. PROCEDURE 7.1. Preparation of BSA standard curve 7.1.1. Take vial of lyophilized BSA standard from BioRad and reconstitute using Milli-Q water to a final concentration of 8mg/ml. 7.1.2. This material is stored in aliquots of 100l at –20oC. 7.1.3. Take 37.5l of the 8mg/ml stock of BSA standard and add 12.5l of milli-Q water for a final volume of 50l and a concentration of 6mg/ml. 7.1.4. Take 25l of the 8mg/ml stock of BSA standard and add 25l of milli-Q water for a final volume of 50l and a concentration of 4mg/ml. 7.1.5. Take 12.5l of the 8mg/ml stock of BSA standard and add 37.5l of milli-Q water for a final volume of 50l and a concentration of 2mg/ml. 7.2. Dilution of unknown protein and BSA standards 7.2.1. Add 800l of milli-Q water into 1ml semi-microcuvettes.

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