Fasciola hepatica – IgG ELISA

Description

Escherichia coli is one of many species of bacteria living in the lower intestines of mammals, known as gut flora. When located in the large intestine, it assists with waste processing, vitamin K production, and food absorption. Discovered in 1885 by Theodor Escherich, a German pediatrician and bacteriologist, E. coli are abundant: the number of individual E. coli bacteria in the faeces that a human defecates in one day averages between 100 billion and 10 trillion. However, the bacteria are not confined to the environment, and specimens have also been located, for example, on the edge of hot springs. The E. coli strain O157:H7 is one of hundreds of strains of the bacterium that causes illness in humans.

E. coli are unable to sporulate. Thus, treatments which kill all active bacteria, such as pasteurization or simple boiling, are effective for their eradication, without requiring the more rigorous sterilization which also deactivates spores. As a result of their adaptation to mammalian intestines, E. coli grow best in vivo or at the higher temperatures characteristic of such an environment, rather than the cooler temperatures found in soil and other environments.

The enteric E. coli (EC) are divided on the basis of virulence properties into enterotoxigenic (ETEC – causative agent of diarrhea in humans, pigs, sheep, goats, cattle, dogs, and horses), enteropathogenic (EPEC – causative agent of diarrhea in humans, rabbits, dogs, cats and horses); enteroinvasive (EIEC – found only in humans), verotoxigenic (VTEC – found in pigs, cattle, dogs and cats); enterohaemorrhagic (EHEC – found in humans, cattle, and goats, attacking porcine strains that colonize the gut in a manner similar to human EPEC strains) and enteroaggregative E. coli (EAggEC – found only in humans).

E. coli O157:H7 was first recognized as a pathogen as a result of an outbreak of unusual gastrointestinal illness in 1982. The outbreak was traced to contaminated hamburgers, and the illness was similar to other incidents in the United States and Japan. The etiologic agent of the illness was identified as a rare O157:H7 serotype of Escherichia coli in 1983. This serotype had only been isolated once before, from a sick patient in 1975.

Exceptional value for money
Rapid detection of all clinically relevant subtypes
Positive copy number standard curve for quantification
Highly specific detection profile
High priming efficiency
Broad dynamic detection range (>6 logs)
Sensitive to < 100 copies of target Accurate controls to confirm findings

Overview

• Adenovirus Purification from any input – cell fraction or media fraction
• Rapid purification within 2 to 4.5 hours
• No specialized equipment needed (ultracentrifuge not required)
• Purify adenovirus cell culture supernatant from 1 mL to 33.5 mL input per prep
• Purify adenovirus cell pellet from 1 mL of input per prep
• Up to 25X sample concentration
• Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix

Adenoviral vectors are useful tools for both in vitro and in vivo gene transfer, and oncolytic viruses based on adenovirus are highly promising for cancer treatment. Norgen’s Adenovirus Purification Kit provides a fast and simple procedure for concentrating and purifying adenoviral vectors from cell lysate and cell culture media. Purification is based on spin column chromatography using Norgen’s proprietary resin as an ion exchanger.  Contaminating cellular debris is largely removed from the sample via a centrifugation step, while contaminating DNA and RNA is reduced using enzymatic digestion. Adenoviral vectors purified in this manner are highly active for use in transduction experiments.

Norgen’s Adenovirus Purification Kit contains sufficient materials for 15 preparations (33.5 mL per prep of supernatant or a total of 500 mL of supernatant input). Approximately 1 mL of cell pellet can be purified per prep, up to a maximum of 15 mL of cell pellet in total for the entire kit. 

Details

Supporting Data

Figure 1 / 3

Previous

Next

Click for expanded view

Kit Specifications
Resin Binding Capacity (total per kit)	At least 3×1011 adenovirus particles as determined by qPCR At least 3×108 transducing units as determined by transduction assay
Input type	Cells, media
Input volume (Supernatant)	1 – 33.5 mL SN per prep (500 mL SN in total)
Input Volume (Cell pellet)	1 mL cell pellet per prep (15 mL in total)
Minimum elution volume	1.3 mL per prep
Time to complete purification	2.5 to 4.5 hours with 1 hour hands on time
In vivo transduction	Yes

Storage Conditions and Product Stability

HL-SAN Nuclease should be stored at -20°C upon arrival. Elution Buffer P should be stored tightly capped at 4°C upon arrival. All other solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment.

Component	Cat. 67600 (15 preps)
Lysis Buffer S	5.5 mL
HL-SAN Nuclease	102 µL
Binding Buffer A	20 mL
Purification Solution C	60 mL
Purification Solution D	130 mL
Wash Solution C	2 x 130 mL
Slurry E	12.5 mL
Elution Buffer P (store at 4°C)	66 mL
Protein Neutralizer	4 mL
Elution tubes (1.7 mL)	50
Product Insert	1

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

Figure 1 / 15

Previous

Next

Click for expanded view

Kit Specifications
Maximum Protein Input	200 μg
Maximum Column Volume Input	600 μL
Molecular Weight of Recovered Proteins	No Molecular Weight cut-off
Final Endotoxin Levels	≤ 0.01 EU/μg protein
Protein Recovery	90-95%
% Detergent Removal	90-95%
Detergents that can be Removed	Including Triton® X-100, CHAPS, NP-40 and Tween 20
Minimum Elution Volume	50 μL
Time to Complete 10 Purifications	20 minutes

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

Component	Cat. 22800 (25 preps)	Cat. 22200 (4 preps)
Binding Buffer J	8 mL	8 mL
Binding Buffer N	4 mL	20 mL
Wash Solution M	50 mL	130 mL
Wash Solution CIP	20 mL	60 mL
Wash Solution N	30 mL	130 mL
Wash Solution NIP	20 mL	60 mL
Elution Buffer G	6 mL	20 mL
Endotoxin Removal Solution	1.5 mL	1.5 mL
Protein Neutralizer EF	2 mL	2 mL
Maxi Spin Columns (assembled with Collection Tubes)	–	4
Mini Spin Columns	25	–
Collection Tubes	25	–
Maxi Spin Columns (assembled with Collection Tubes)	4
Elution Tubes (1.7 mL)	25	–
Elution Tubes (50 mL)	4	–
Product Insert	1	1