Overview

• Columns bind proteins of interest while endotoxins flow through
• Reduce endotoxin levels to less than 0.01 EU/µg of protein
• Proteins are desalted
• Greater than 95% protein recovery
• Concentrate protein samples and remove detergents at the same time
• Effectively remove a wide range of detergents including SDS, Triton® X-100, CHAPS, NP-40, and Tween 20
• Purification is based on spin column chromatography that uses Norgen’s resin separation matrix

The ProteoSpin™ Total Protein Concentration, Detergent Clean-Up and Endotoxin Removal Kits are designed for the rapid removal of endotoxins from previously purified proteins or peptides, with protein recoveries of > 95% being achieved. Endotoxins, also known as lipopolysaccharides, are cell-membrane components of Gram-negative bacteria such as E. coli. Endotoxins liberated by Gram-negative bacteria are frequent contaminations of protein solutions derived from bioprocesses. Due to the high toxicity of endotoxins in vivo and in vitro, their removal from protein preparations is often necessary prior to the use of the protein in downstream applications. These kits efficiently reduce endotoxin levels to ≤ 0.01 EU/mg of protein, using spin column chromatography based on Norgen’s proprietary resin as the separation matrix. These kits are highly efficient in removing many different salts commonly used in the laboratory including, but not limited to, MgCl₂, NaCl, KCl, CaCl₂, LiCl and CsCl. The purified protein samples can be used in a number of downstream applications including sequencing, cloning, and in vitro and in vivo introduction into cells and organisms for various purposes. The simultaneous removal of salts while concentrating a dilute protein solution makes the kit a convenient method for preparing proteins before running many downstream applications such as SDS-PAGE, isoelectric focusing, X-ray crystallography, NMR spectroscopy, mass spectroscopy and other applications.

Mini Kit

The ProteoSpin™ Total Protein Concentration, Detergent Clean-Up and Endotoxin Removal Mini Kit is designed for the rapid removal of endotoxins from up to 200 μg of previously purified proteins or peptides, with protein recoveries of > 95% being achieved.

Maxi Kit

The ProteoSpin™ Total Protein Concentration, Detergent Clean-Up and Endotoxin Removal Maxi Kit is designed for the rapid removal of endotoxins from up to 4 mg of previously purified proteins or peptides, with protein recoveries of > 95% being achieved.

Details

Supporting Data

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Storage Conditions
All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment.

Description

Specifications

PAX-8 is a member of the paired box (PAX) family of transcription factors, which are key regulators in early development. This protein plays a role in development of thyroid follicular cells and the expression of thyroid-specific genes, with mutations in the PAX-8 gene linked to thyroid follicular carcinomas, atypical thyroid adenomas, and thyroid dysgenesis. The PAX-8 protein is expressed in simple ovarian inclusion cysts and non-ciliated mucosal cells of the fallopian tubes, but is absent from normal ovarian surface epithelial cells. PAX-8 is also not expressed in normal lung or lung carcinomas. Reports have associated PAX-8 expression with renal carcinoma, nephroblastoma, and seminoma, and have indicated PAX-8 as a useful marker for renal epithelial tumors, ovarian cancer, and for differential diagnoses in lung and neck tumors. Anti-PAX-8 can be useful in determining the primary site of invasive micropapillary carcinomas of ovary from bladder, lung, and breast, when used in adjunct with a panel of organ-specific markers such as uroplakin, mammaglobin, and TTF-1.

Introduction

MagZol Reagent is a reagent system for the isolation of total RNA from cells and tissues. The reagent, a single-phase solution consisting of phenol and guanidine isothiocyanate, is modification of the single-step RNA isolation method developed by Chomczynski and Sacchi. The sample is homogenized and lysed in MagZol Reagent which maintains the integrity of the RNA, while disrupting and denaturing endogenous RNases and other cellular components. Extraction of the lysate with chloroform further denatures proteins and separates the mixture into an organic and an aqueous phase. RNA remains exclusively in the aqueous phase, and is subsequently recovered by isopropanol.

This method is suitable for small quantities of tissue (<100mg) and cells (<5 X10⁶), and large quantities of tissue (up to 1g) and cells (<10⁸), of human, animal, plant, or bacterial origin. The simplicity of the MagZol Reagent method allows simultaneous processing of a large number of samples. The entire procedure can be completed in one hour. Total RNA prepared in this manner can be used for Northern blot analysis, dot blot hybridization, poly(A) + selection, in vitro translation, RNase protection assay, and molecular cloning. For use in amplification by thermal cycling, treatment of the isolated RNA with RNase-free DNase I is recommended when the two amplimers lie within a single exon.

Details

Specifications

Advantages

• Flexible – sample amount can be adjusted according to the demand
• Cost performance -the most economical nucleic acid extraction technology

Storage and Stability

MagZol Reagent should be stored at 2-8°C upon arrival and is stable for at least 24 months under the condition. However, short-term storage (up to 12 weeks) at room temperature (15-25°C) does not affect its performance.

MagZol Reagent is a reagent system for the isolation of total RNA from cells and tissues. The reagent, a single-phase solution consisting of phenol and guanidine isothiocyanate, is modification of the single-step RNA isolation method developed by Chomczynski and Sacchi. The sample is homogenized and lysed in MagZol Reagent which maintains the integrity of the RNA, while disrupting and denaturing endogenous RNases and other cellular components. Extraction of the lysate with chloroform further denatures proteins and separates the mixture into an organic and an aqueous phase. RNA remains exclusively in the aqueous phase, and is subsequently recovered by isopropanol.