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Rapid Isolation of Total Protein from Arabidopsis Pollen.

Bio-protocol (2017-04-20)
Ming Chang, Shanjin Huang
ABSTRACT

Arabidopsis pollen is an excellent system for answering important biological questions about the establishment and maintenance of cellular polarity and polar cell growth, because these processes are amenable to the genetic and genomic approaches that are readily available in Arabidopsis. Given that proteins are the direct executors of a wide variety of cellular processes, it is important to rapidly and efficiently isolate total protein for various protein-based analyses, such as Western blotting, co-immunoprecipitation and mass spectrometry, among others. Here we present a protocol for rapid isolation of total protein from Arabidopsis pollen, which is adapted from our recently published paper (Chang and Huang, 2015).

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Ponceau S, BioReagent, suitable for electrophoresis
Sigma-Aldrich
Phenylmethylsulfonyl Fluoride, Phenylmethylsulfonyl Fluoride, CAS 329-98-6, is an irreversible inhibitor of serine proteases. It causes sulfonylation of the active-site serine residues.