Isolate high quality DNA from a broad variety of phage strains
High yields of total DNA
Fast and easy processing using a rapid spin-column format
No phenol or chloroform extractions or cesium chloride banding required
High yields of DNA recovered3-15 µg DNA from 106-1010 pfu/ mL of enriched phages
This kit provides a rapid spin column method for the purification of total DNA from a broad spectrum of bacteriophages propagated in bacteria grown in liquid cultures. The DNA is isolated without the use of phenol, chloroform or cesium chloride banding procedures. The spin-column based procedure is rapid and can be completed in less than 45 minutes. The kit is highly efficient for processing small volumes of phage supernatant (500 µL – 1 mL) and with the optional DNase and Proteinase K treatments phage DNA yields are maximized while host DNA contamination is minimized. Purified total phage DNA is of the highest integrity, and can be used in a number of downstream applications including PCR, qPCR, Restriction Fragment Length Polymorphism (RFLP), sequencing, cloning, Southern Blot and more.
3-15 µg DNA from 106-1010 pfu/mL of enriched phages
Time to Complete 10 Purifications
45 minutes
* Average yields will vary depending upon a number conditions used and developmental stage.
Storage Conditions and Product Stability All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 1 year after the date of shipment.
Component
Cat. 46800 (50 preps)
Cat. 46850 (100 preps)
Lysis Buffer B
40 mL
2 x 40 mL
Wash Solution A
38 mL
2 x 38 mL
Elution Buffer B
8 mL
2 x 8 mL
Spin Columns
50
100
Collection Tubes
50
100
Elution Tubes (1.7 mL)
50
100
Product Insert
1
1
Other Products
Fasciola hepatica – IgG ELISA
Product Info
Document
Product Info
Name of Product
Fasciola hepatica – IgG ELISA
Catalog Number
AF 9650
Short Info
Fasciolosis is one of the most frequently encountered autochthonous helminthic infections in Central Europe. The immunodiagnosis of Fasciola hepatica infections is challenged by high serological cross-reacitivity with other (hepatic) parasitological infections encountered in Central Europe, such as alveolar echinococcosis, toxocarosis and ascariosis, but also other parasitic infections acquired during overseas travel. The SAP-2 recombinant antigen shows a high specificity, especially with patients with other parasitic infections. It is a suitable serological test for routine diagnosis of human fasciolosis, particularly if the results are supported by clinical history.
This product is manufactured by Bordier Affinity Products in Switzerland and distributed in Germany exclusively by Milenia Biotec.
Method/Platform
ELISA in microplate format
Range/Assay Sensivity
77% sensitivity, 98% specificity
Test Principle
The kit provides all the material needed to perform 96 enzyme-linked immunosorbant assays (ELISA) on breakable microtitration wells sensitized with Fasciola hepaticarecombinant antigen. Specific antibodies in the sample will bind to the antigen and washing will remove unspecific antibodies. The presence of parasite specific antibodies is detected with a Protein A – alkaline phosphatase conjugate. A second washing step will remove unbound conjugate. Revealing bound antibodies is made by the addition of pNPP substrate which turns yellow in the presence of alkaline phosphatase. Color intensity is proportional to the amount of Fasciola hepatica specific antibodies in the sample. Potassium phosphate is added to stop the reaction. Absorbance at 405 nm is read using an ELISA microplate reader.The test can be performed with automatic systems, but this must be validated by the user.
12 x 8 strips (96 tests)
Fasciolosis is one of the most frequently encountered autochthonous helminthic infections in Central Europe. The immunodiagnosis of Fasciola hepatica infections is challenged by high serological cross-reacitivity with other (hepatic) parasitological infections encountered in Central Europe, such as alveolar echinococcosis, toxocarosis and ascariosis, but also other parasitic infections acquired during overseas travel. The SAP-2 recombinant antigen shows a high specificity, especially with patients with other parasitic infections. It is a suitable serological test for routine diagnosis of human fasciolosis, particularly if the results are supported by clinical history.
Apoptosis is an essentially normal physiological process that removes now redundant, cells, particularly during embryonic development and early growth. In adult animals the process removes cells that are irreparable. The apoptotic process is also involved in many major diseases such as cancer, where transformed tumour cells have their apoptotic process disabled, permitting cell cycling to continue unchecked. In contrast some forms of senile dementia may result from excessive apoptotic induction of neural cells.
The apoptotic process in mammalian cells is a rapid event (2‐4 hours). Within this short time span an apparently viable cell can be quietly dismantled, to disappear leaving no visible trace of its former existence.
How is apoptosis detected or measured?
An apoptosis cascade of activators, effectors and regulators has been identified. This in turn led to a range of apoptosis assays being devised to detect and monitor these events. Some laboratories will employ two distinct assays, one selected to detect early (initiation) apoptotic events, while a second assay will target a later (execution) event. Apoptosis assays, based on methodology, can be classified into four major inter‐linked groups:
[1] DNA fragmentation (electrophoresis and nick end labelling, TUNEL).
[2] Apoptotic proteases (fluorescently labelled antibodies to the caspases).
[3] Flow cytometric analysis (FACS, incorporating other group assays).
Biocolor’s APOPercentage assay is based on the latter. Further information can be found under the ‘Mode of Action’ Tab.
How does APOPercentage detect apoptosis?
The mammalian cell membrane has been described as a semi‐fluid mosaic structure, composed of phospholipids with a diverse group of inserted proteins and some cholesterol. The phospholipids are the major components of the membrane and are arranged in the form of a ‘bi‐layer’; which is asymmetric in composition, structure, and function.
To ensure normal transmembrane functions the phospholipids must be maintained in an asymmetric composition. The process is regulated by ‘flippases’, which catalyse the active transport of aminophospholipids from the outer to inner monolayer. However, in cells undergoing apoptosis, flippase is overwhelmed by the action of another enzyme, termed ‘floppase’ or ‘scramblase’. The net effect is a scrambling of the phospholipid distribution between the inner and outer monolayers.
Cell membrane changes during apoptosis
The APOPercentage assay utilises an intense, pink-coloured dye reagent which is taken up during in-vitro culture by apoptosis-committed cells. This uptake occurs at the stage of Phosphatidylserine transmembrane movement, as produced by the flipflop mechanism. Dye uptake continues until blebbing occurs. No further dye can then enter the now defunct cell and the dye that has accumulated within the cell is not released (unlike necrotic cells which release dye).
Since the dye reagent is excluded or not retained by healthy or necrotic cells it therefore acts as a specific label for apoptotic cells.
How are APOPercentage-labelled cells quantified?
Labelled apoptosis cells may then by conveniently analysed by the following methods:
Direct Analysis The intense pink colour of the labelled cells can be visually assessed using brightfield microscopy. Apoptosis in substrate-adherent cell populations is therefore readily quantified using image analysis techniques. This technique is the most sensitive with the ability of detecting one single apoptotic cell per well.
Colorimetry protocol Dye that accumulates within apoptotic cells is released into solution via addition of Dye Release Reagent. The concentration of this intracellular dye is then measured at 550nm using a microplate colorimeter/spectrophotometer.
NB: The APOPercentage assay kit does NOT require the use of a Flow Cytometer.
Limit of Detection
A single cell (via image analysis method)
Detection Method
Colorimetric (550nm) (Endpoint) or Image Analysis based
Measurements per kit
Sufficient for 4×24 well plates or 6×96 well plates
Suitable Samples
Adherent mammalian cells (in-vitro)
APOPercentage kit contents:
1. APOPercentage Dye (1x5ml)
2. Dye Release Reagent (1x150ml)
3. Phosphate Buffered Saline (PBS) (1x120ml)
4. 24-well starter plate.
5. Assay kit manual.
The Colorimetric Protocol requires a Microplate Colorimeter / Spectrophotometer.
Additional 96-well plates will be required for use when reading dye absorbance values.
The Direct Detection Protocol Requires an inverted stage microscope with an attached digital camera.
NB: Additional reagents (typically culture medium and suitable apoptosis treatments) may be required for sample preparation prior to assay. Consult manual or contact us for further details.
Document
The APOPercentage™ Apoptosis kit is a dye-based, colorimetric assay for detection and measurement of apoptosis (programmed cell death) during in-vitro cell culture.
High yield and high quality genomic DNA with no RNA or protein contamination
DNA ready for any application including PCR, qPCR, genotyping and more
Norgen’s Cells and Tissue DNA Isolation Kits are designed for the rapid preparation of genomic DNA from cultured cells as well as various tissue samples and urine. The purified genomic DNA is fully digestible with all restriction enzymes tested, and is completely compatible with PCR and Southern Blot analysis.
Cells and Tissue DNA Isolation Kit (Spin Column)
Purification is based on spin column chromatography as the separation matrix. Norgen’s columns bind DNA under optimized salt concentrations and release the bound DNA under low salt and slightly alkali conditions. The protocol can be completed in approximately 30 minutes for cells and within 90 minutes for tissues. Each kit contains sufficient materials for 50 preparations.
Cells and Tissue DNA Isolation Micro Kit (Micro)
Optimized for small inputs of cells and tissues, such as Laser-Captured Microdissection (LCM). Purification is based on spin column chromatography as the separation matrix. Norgen’s columns bind DNA under optimized salt concentrations and release the bound DNA under low salt and slightly alkali conditions. Preparation time for a single sample is approximately 60 minutes, and each kit contains sufficient materials for 50 preparations.
Cells and Tissue DNA Isolation Kit (Magnetic Bead System)
Purification is based on the use of magnetic beads that bind DNA under optimized binding conditions. Norgen’s Cells and Tissue DNA Isolation Kit (Magnetic Bead System) allows for the isolation of genomic DNA from various types of animal tissues or cell samples. Preparation for 10 purifications is approximately 40 minutes of hands-on time.
Cells and Tissue DNA Isolation 96-Well Kit (High Throughput Magnetic Bead System)
Purification is based on the use of magnetic beads that bind DNA under optimized binding conditions. Norgen’s Cells and Tissue DNA Isolation 96-Well Kit (Magnetic Bead System) allows for the isolation of genomic DNA from various types of animal tissues or cell samples. The Cells and Tissue DNA Isolation 96-Well Kit (Magnetic Bead System) also can be integrated with a robotic automation system.
8-10 µg (20 mg of animal tissue) 8-12 µg (3 x 106 cells)
Average Purity (OD260/280)
1.8 – 1.9
Time to Complete 10 Purifications
40 minutes hands-on time (Cat. 59100) 50 minutes hands on time (Cat. 62500)
* Average DNA yield will vary depending on the donor
Storage Conditions and Product Stability All solutions should be kept tightly sealed and stored at room temperature (15 – 25°C). This kit is stable for 1 year after the date of shipment.
