Detergent-Free Total Protein Isolation Kit

Overview

Simple protocol that is free of detergents
Fast processing time
Process a variety of inputs
Compatible with mass spectrometry and NMR spectroscopy
Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix

This kit provides a fast and simple procedure for the isolation of total proteins from tissue, bacteria, yeast or mammalian cells without the use of SDS, Triton® X-100 and other detergents. Detergents are extensively used to prepare protein samples; however, these detergents have undesirable effects on downstream analysis. These effects include extraneous peaks in mass spectrometry, artifacts with chromatography and electrophoresis, interference with microinjection into cells and interference with protein immunization.

The Detergent-Free Total Protein Isolation Kit maintains high protein recovery and yields proteins that are 100% detergent-free. Purification is based on using Norgen’s proprietary resin together with Lysis Solution, followed by protein filtration using a filter column (provided). The purified proteins can be used in a number of downstream applications including mass spectrometry, SDS-PAGE, isoelectric focusing, NMR spectroscopy and more.

Each Lysis Tube is able to process up to 50 mg of tissue, 10¹⁰ bacterial cells, 10⁹ yeast cells or 10⁷ mammalian cells. Preparation time for 12 samples is less than 10 minutes.

Details

Supporting Data

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Kit Specifications
Maximum Amount Of Starting Material: Tissues Animal Cells Bacteria Yeast	50 mg 1 x 10⁷ cells 1 x 10¹⁰ cells 1 x 10⁹ cells
Time to Process 12 Samples	Less than 10 minutes

Storage Conditions and Product Stability
The Lysis Solution should be kept tightly sealed and stored at room temperature. Once opened, the solution should be stored at 4°C. This kit is stable for 2 years after the date of shipment.

Component	Cat. 30300 (50 preps)
Lysis Solution	4 mL
Lysis Tubes	25
Filter Columns	25
Elution Tubes (1.7 mL)	25
Product Insert	1

Content:	100 assays (manual) / 1000 assays (microplate) / 1000 assays (auto-analyser)
Shipping Temperature:	Ambient
Storage Temperature:	Short term stability: 2-8^oC, Long term stability: See individual component labels
Stability:	> 2 years under recommended storage conditions

Analyte:	D-Isocitric Acid
Assay Format:	Spectrophotometer, Microplate, Auto-analyser
Detection Method:	Absorbance
Wavelength (nm):	340
Signal Response:	Increase
Linear Range:	1 to 80 µg of D-isocitric acid per assay
Limit of Detection:	0.35 mg/L
Reaction Time (min):	~ 3 min
Application examples:	Fruit juices, fruit products, soft drinks and other materials (e.g. biological cultures, samples, etc.).
Method recognition:	Methods based on this principle have been accepted by EN, NEN, NF, DIN, GOST, IFU and AIJN

Kit Specifications
Column Binding Capacity	25 µg
Size of Plasmids Purified	Up to 13,000 bp
Average Yield from 1.5 mL of Culture	Up to 20 µg
Time to Complete 10 Purifications	30 minutes

Component	Cat. 13300 (50 preps)	Cat. 46400 (250 preps)	Cat. 46500 (4 preps)	Cat. 46600 (20 preps)	Cat. 60300 (50 preps)	Cat. 63000 (192 preps)
Resuspension Solution AZ	12 mL	60 mL	20 mL	100 mL	12 mL	2 x 20 mL
Lysis Buffer N	40 mL	80 mL	40 mL	2 x 80 mL	40 mL	2 x 40 mL
Buffer TN	20 mL	130 mL	55 mL	2 x 130 mL	20 mL	1 x 55 mL 1 x 20 mL
Wash Solution E	12 mL	2 x 18 mL	–	–	–	–
Elution Buffer K	8 mL	30 mL	–	–	8 mL	2 x 8 mL
Wash Solution J	–	–	25 mL	3 x 25 mL	–	–
Elution Buffer J	–	–	24 mL	120 mL	–	–
RNase A	1 vial	1 vial	1 vial	1 vial	1 vial	1 vial
Magnetic Bead Suspension	–	–	–	–	1 x 1.1 mL	4 x 1.1 mL
Spin Columns	50	250	–	–	–	–
Collection Tubes	50	250	–	–	–	–
DNA Maxi Spin Columns with Collection Tubes (Clear ring in column)	–	–	4	20	–	–
Maxi Spin Filter Columns with Collection Tubes (Grey ring in column)	–	–	4	20	–	–
96-Well Plate	–	–	–	–	–	2
Elution Tubes (1.7 mL)	50	250	–	–	50	–
Elution Tubes (50 mL)	–	–	4	20
96-Well Elution Plate	–	–	–	–	–	2
Adhesive Tape	–	–	–	–	–	2
Product Insert	1	1	1	1	1	1

Detergent-Free Total Protein Isolation Kit

Detail

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Supporting Data

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