PE0240

Sigma-Aldrich

Plant Fractionated Protein Extraction Kit

Suitable for any plant species or tissue

NACRES:
NA.56

Quality Level

usage

sufficient for 20 extractions

storage temp.

−20°C

General description

Sigma′s Plant Fractionated Protein Extraction Kit is designed specifically for use in plant bioscience to extract qualitative hydrophilic and hydrophobic proteins samples from any type of plant species or tissue for downstream proteomic applications. The protocol does not require any ultracentrifugation or aqueous polymer two-phase partitioning (APTP). The kit includes five reagents; a plant specific protease inhibitor, a specially formulated reagent to extract hydrophilic proteins and a new chaotropic reagent with increased solubilizing power to extract more hydrophobic proteins. Also included in the kit are reducing reagent Tributylphosphine (TBP) and alkylating reagent, Iodoacetamide. These reagents improve separation during isoelectric focusing and 2-D gel electrophoresis. The protease inhibitor is a mixture of protease inhibitors with broad specificity for the inhibition of serine, cysteine, aspartic, metalloproteases and aminopeptidases. The cocktail contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF), bestatin, pepstatinA, E-64, leupeptin, and 1,10-phenanthroline.
Following removal of polyphenolics, tannins and other interfering substances, ground plant tissue, fresh or frozen, is resuspended in a specially formulated reagent to extract hydrophilic proteins. After sequential extractions of hydrophilic proteins, addition of the chaotropic reagent provided extracts hydrophobic membrane bound proteins. Plant debris is pelleted by centrifugation and protein extract solutions are collected. The end results are qualitative fractionated protein samples, ready for downstream proteomic analysis.

Kit Components Also Available Separately

Product No.
Description
SDS

  • Plant Protein Extraction Reagent Type 1 1 bottle

  • C0356Protein Extraction Reagent Type 4 1 bottle

  • P9599Protease Inhibitor Cocktail, for plant cell and tissue extracts, DMSO solution 3 x 1

  • T7567Tributylphosphine solution, 200 mM (in N-methyl-2-pyrrolidinone), liquid 3 x 0.5

  • A3221Iodoacetamide, Single use vial of 56 mg 3 vial(s)

Signal Word

Danger

hazcat

Acute Tox. 3 Oral - Aquatic Chronic 2 - Carc. 2 - Eye Dam. 1 - Repr. 1B - Self-heat. 1 - Skin Corr. 1B - Skin Sens. 1 - STOT SE 3

Target Organs

Respiratory system

storage_class_code

4.2 - Pyrophoric and self-heating hazardous materials

Certificate of Analysis

Certificate of Origin

Loomis, W.D.
Methods in Enzymology, 31, 528-545 null
M Ferro et al.
Electrophoresis, 21(16), 3517-3526 (2000-11-18)
As a complementary approach to genome projects, proteomic analyses have been set up to identify new gene products. One of the major challenges in proteomics concerns membrane proteins, especially the minor ones. A procedure based on the differential extraction of...
Yinghui Ying et al.
Plant physiology, 173(1), 812-824 (2016-11-30)
Phosphate overaccumulator2 (PHO2) encodes a ubiquitin-conjugating E2 enzyme that is a major negative regulator of the inorganic phosphate (Pi)-starvation response-signaling pathway. A yeast two-hybrid (Y2H) screen in rice (Oryza sativa; Os) using OsPHO2 as bait revealed an interaction between OsPHO2...
Wenhao Yue et al.
The Plant journal : for cell and molecular biology, 90(6), 1040-1051 (2017-02-24)
Inorganic phosphate (Pi) transporters (PTs) play vital roles in Pi uptake and translocation in plants. Under Pi sufficient conditions, PTs are degraded to prevent excess Pi accumulation. The mechanisms targeting PTs for degradation are not fully elucidated. In this study...
Angus L Dawe et al.
Methods in molecular biology (Clifton, N.J.), 722, 225-236 (2011-05-19)
The interaction of the filamentous fungal plant pathogen Cryphonectria parasitica with its virulence-attenuating viruses provides a unique platform to explore the molecular biology and genetics of virus-host interactions. Following the development of transformation procedures for this fungus, subsequent advances include...

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