Reagenti per estrazione di proteine
Una lisi cellulare efficiente e rese massime di estrazione proteica sono vitali per una purificazione di proteine ricombinanti di alta qualità. I cocktail di inibitori delle proteasi sono utilizzati per lisare cellule di mammifero, tamponi che solubilizzano le membrane plasmatiche e intracellulari, rompono i deboli legami intermolecolari e solubilizzano la maggior parte delle proteine. Questi reagenti di estrazione proteica, che rendono le procedure più facili da eseguire, possono anche essere utilizzati per isolare proteine dalle frazioni citoplasmatiche e nucleari di cellule coltivate.
Ovalbumin (in PBS), Imject™
Supplier: Thermo Fisher Scientific
Ovalbumin (in PBS), Imject™
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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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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 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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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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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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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, 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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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 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
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)
Il reagente PopCulture® è un concentrato a base di detergente che può essere aggiunto direttamente alle colture di E. coli per estrarre efficacemente proteine senza la necessità della raccolta di cellule. Le proteine ricombinanti possono sottoposte a screening direttamente nell'estratto grezzo oppure purificate mediante l'aggiunta di una matrice di affinità, il lavaggio del complesso matrice-proteine target per rimuovere il terreno di coltura esaurito e i contaminanti cellulari, nonché l'eluizione della proteina purificata dalla matrice. La procedura di purificazione delle proteine PopCulture è ideale per l'elaborazione robotica ad alta resa di campioni per la ricerca proteomica o tutte le applicazioni che trarrebbero vantaggio dal miglioramento di velocità e praticità.
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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™
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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