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Réactifs pour l'extraction des protéines

Réactifs pour l'extraction des protéines

Une lyse cellulaire efficace et un rendement maximal d’extraction des protéines sont essentiels à une purification de protéines recombinantes de haute qualité. Les cocktails d’inhibiteurs de protéases servent à lyser les cellules de mammifères, avec des tampons qui solubilisent les membranes plasmiques et intracellulaires, rompent les faibles liaisons intermoléculaires et solubilisent la plupart des protéines. Ces réactifs d’extraction de protéines, qui facilitent les procédures, peuvent également être utilisés pour isoler des protéines des fractions cytoplasmiques et nucléaires de cellules cultivées.

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Ovalbumin (in PBS), Imject™

Ovalbumin (in PBS), Imject™

Supplier: Thermo Fisher Scientific

Ovalbumin (in PBS), Imject™

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Immobilised Papain

Immobilised Papain

Supplier: G-Biosciences

A cysteine protease enzyme (EC 3.4.22.2) immobilised on 4% agarose, cleaves Immunoglobulin G (IgG) antibody molecules in the hinge region, generating three ~50 kDa fragments; two Fab domains and a Fc domain. The papain-digested antibody is unable to promote agglutination, precipitation, opsonisation, and lysis.

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Micrococcal nuclease solution (≥100 U/µl)

Micrococcal nuclease solution (≥100 U/µl)

Supplier: Thermo Fisher Scientific

This Micrococcal Nuclease (MNase) is suitable for removing nucleic acids from cell lysates, releasing chromatin-bound proteins and shearing chromatin for use in chromatin immunoprecipitation (ChIP) experiments.

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Immobilised Pepsin

Immobilised Pepsin

Supplier: G-Biosciences

A proteolytic enzyme immobilised on 4% agarose that is routinely used for the generation of F(ab’)₂ fragments from Immunoglobulin G (IgG). The pepsin has the ability to cleave the heavy chains near the hinge region. The immobilised Pepsin offers the distinct advantage of eliminating enzyme contamination of the F(ab)₂ fragments.

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SDS precipitation kit, SDS-Out™

SDS precipitation kit, SDS-Out™

Supplier: Thermo Fisher Scientific

SDS-Out™ SDS Precipitation Kit provides convenient removal of SDS from small-volume (0,1 to 10 ml) protein samples.

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Immobilised trypsin, TPCK treated, Pierce™

Immobilised trypsin, TPCK treated, Pierce™

Supplier: Thermo Fisher Scientific

Pierce™ immobilised TPCK trypsin is a serine endoprotease applicable to amino acid analysis and protein sequencing, mapping and structural studies. The immobilised form allows sample separation after treatment.

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Mass tagging, mass tag labelling reagents, AminoxyTMT™, Pierce™

Mass tagging, mass tag labelling reagents, AminoxyTMT™, Pierce™

Supplier: Thermo Fisher Scientific

The carbonyl-reactive Thermo Scientific™ aminoxyTMT™ (Tandem Mass Tag™) Label Reagents enable multiplexed characterisation and quantitation of carbonyl-containing biomolecules (carbohydrates, steroids, oxidised proteins) by mass spectrometry (MS).

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Immobilised Trypsin

Immobilised Trypsin

Supplier: G-Biosciences

Trypsin is a serine endopeptidase that specifically cleaves peptide bonds on the carboxy side of s-aminoethyl cysteine, arginine and lysine residues and typically there is little or no cleavage at arginyl-proline and lysyl-proline bonds. The distribution of these residues in proteins allows trypsin digestion to produce peptides that are readily identified by mass spectrometry.

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Detergent sampler Kit, Surfact-Amps™

Detergent sampler Kit, Surfact-Amps™

Supplier: Thermo Fisher Scientific

Surfact-Amps™ Detergent sampler is a collection of various Surfact-Amps™ detergent solutions and selected other useful detergents for protein research methods.

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Detergent solutions, Surfact-Amps™

Detergent solutions, Surfact-Amps™

Supplier: Thermo Fisher Scientific

Surfact-Amps™ detergents are ready-to-use, diluted solutions of purified detergent that can be used in routine and high-demand protein research methods and molecular biology techniques. These detergents are useful as a component of cell lysis buffers or other solutions intended to extract and solubilise proteins, improve chromatography, prepare yeast spheroplasts, and reduce background in ELISA or Western blot methods.

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Lysing matrix Z, MP Biomedicals

Lysing matrix Z, MP Biomedicals

Supplier: MP Biomedicals

Lysing Matrix Z is used primarily for lysis of animal and plant tissue, tougher stems, roots and seeds, and whole insects. Each impact-resistant tube contains yttria-stabilised zirconium oxide beads.

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Endoproteinase Glu-C, excision grade, Staphylococcus aureus

Supplier: Merck Millipore (Calbiochem‎)

Native endoproteinase Glu-C from Staphylococcus aureus. Serine protease that specifically hydrolyses peptide bonds at the carboxylic side of glutamic acid and aspartic acid residues. Inhibited by DFP and α2-macroglobulin. Suggested working concentration: 1:20 to 1:100 (protease:protein by weight) for sequence analysis.

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PopCulture® reagent

Supplier: Merck Millipore (Novagen)

Le réactif PopCulture® est un concentré à base de détergent qui peut être ajouté directement aux cultures d'E. coli pour une extraction efficace des protéines sans récolte de cellules. Les protéines recombinantes peuvent être directement criblées dans l'extrait brut ou purifiées par l'ajout d'une matrice d'affinité, le lavage du complexe matrice-protéines cibles afin d'en éliminer le milieu de culture usagé et les contaminants cellulaires et l'élution des protéines purifiées de la matrice. La procédure de purification de protéines PopCulture convient parfaitement au traitement robotisé haut débit d'échantillons pour la recherche protéomique ou pour toute autre application pouvant bénéficier de l'accélération et du confort accru des procédures.

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Trypsin protease, MS grade

Trypsin protease, MS grade

Supplier: Thermo Fisher Scientific

Pierce™ Trypsin Protease, MS Grade, is highly purified porcine trypsin that has been chemically modified for maximum activity and stability in proteomic applications.

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Master mix, BugBuster®

Supplier: Merck Millipore (Novagen)

BugBuster® master mix combines BugBuster® protein extraction reagent with Benzonase® Nuclease and rLysozyme™ solution in one convenient reagent. It allows for maximum recovery of active soluble protein from both Gram-negative and Gram-positive bacteria. The master mix provides sufficient reagents for protein extraction from 20 g and 100 g cell paste.

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Protein extraction reagents, CytoBuster™

Protein extraction reagents, CytoBuster™

Supplier: Merck Millipore (Novagen)

The CytoBuster™ protein extraction reagent is a proprietary formulation of detergents optimised for efficient extraction of soluble proteins from mammalian and insect cells. The gentle, non-ionic composition of the reagent enables isolation of functionally active endogenous or expressed proteins without a need for secondary treatment such as sonication or freeze/thaw.

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