CE-IVDR marked in accordance with the European Commission Regulation (EU) No. 2017/746.
Ideal for use in in vitro diagnostic workflows
Simultaneous isolation of both host DNA and microbial DNA (universal protocol)
Eliminates PCR inhibitors including humic acids
Fully compatible with Norgen’s Stool Nucleic Acid Collection and Transport Tubes
High quality DNA for sensitive downstream applications including PCR, qPCR, sequencing and microarray
This kit provides a convenient and rapid method to isolate total DNA from fresh, frozen and preserved stool samples, including those preserved using Norgen’s Stool Nucleic Acid Collection and Preservation Devices Dx (Cat. Dx45660). The universal protocol conveniently allows for the isolation of total genomic DNA from all the various microorganisms and host cells found in the stool sample simultaneously. The kit removes all traces of humic acids and other inhibitors using Bead Tubes and a combination of chemical and physical homogenization and lysis, without the need of any phenol-chloroform extractions. A simple and rapid spin column procedure is then used to further purify the DNA with high yields and molecular weights of up to 50 kb plus. The purified DNA is of the highest quality and is fully compatible with all downstream applications such as PCR, qPCR, NGS and microarrays since all humic acid substances and other PCR inhibitors are removed during the isolation process.
NOTE: This product is not available for sale in the United States.
200 mg (fresh/frozen stool) or 400 μL (preserved stool)
Type of Stool Processed
Frozen, fresh or preserved stool
Format
Spin Column
Maximum Column Binding Capacity
50 μg
Maximum Column Loading Volume
650 μL
Elution Volume
50 μL
Time to Complete 10 Purifications
30 minutes
Applications
PCR, qPCR, Southern Blot Analysis, Sequencing, Microarray Analysis.
Storage Conditions and Product Stability All 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. Dx27600 (50 preps)
Lysis Solution
60 mL
Lysis Additive
6 mL
Binding Solution
7 mL
Wash Solution I
30 mL
Wash Solution II
22 mL
Elution Buffer
3 mL x 2
Bead Tube
50
Mini Spin Columns
50
Collection Tubes
50
Elution Tubes (1.7 mL)
50
Product Insert
1
Other Products
APOPercentage apoptosis assay kit
Product Info
Document
Product Info
What is Apoptosis?
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.
PEG13-bis(Amino-Tri-(Propargyl-PEG2-ethoxymethyl)-methane) is large molecule with two sets of three terminal alkynes joined together by a large PEG linker. The alkynes are most frequently used in copper click chemistry with azides. The PEG linkers as well as numerous amide bonds provide high aqueous solubility to this compound, which may alter ADME of this compound.
Document
PEG13-bis(Amino-Tri-(Propargyl-PEG2-ethoxymethyl)-methane) is large molecule with two sets of three terminal alkynes joined together by a large PEG linker. The alkynes are most frequently used in copper click chemistry with azides. The PEG linkers as well as numerous amide bonds provide high aqueous solubility to this compound, which may alter ADME of this compound.
Process a broad spectrum of plant species and filamentous fungi
Isolate total DNA including pathogen DNA without phenol
Available in spin column format and 96-well format for high throughput applications
These kits provide a rapid method for the isolation and purification of total DNA from a wide range of plant and fungal species. Total DNA, including genomic DNA, mitochondrial DNA and chloroplast DNA can be purified from fresh or frozen plant tissues, plant cells or fungi samples using this kit. Purified DNA samples can be used for the detection of viral pathogens, as viral DNA is isolated with the plant/fungi DNA. The purified DNA is of the highest integrity, and can be used in a number of downstream applications including PCR, qPCR, SNP, Southern blotting and sequencing.
Plant/Fungi DNA Isolation Kit (Spin Column)
Complete 10 purifications in 45 minutes. This kit offers a maximum loading volume of 650 μL per column, and a maximum binding capacity of 50 μg per column.
Plant/Fungi DNA Isolation 96-Well Kit (High Throughput)
For high throughput applications. Purification with the 96-well plates can be integrated with a robotic automation system. Complete 96 purifications in 50 minutes. This kit offers a maximum loading volume of 500 μL per well, and a maximum binding capacity of 50 μg per well.
Plant/Fungi DNA Isolation Kit (Magnetic Bead System)
The DNA is bound to the surface of the magnetic beads under optimized buffer conditions and released using a low salt buffer system. The Plant DNA Isolation Kit (Magnetic Bead System) can be easily adapted to automated magnetic bead separation instruments and work stations.
Plant/Fungi DNA Isolation 96-Well Kit (High Throughput Magnetic Bead System)
For high throughput applications. Purification with the 96-well plates can be integrated with a robotic automation system.
* Average yields will vary depending upon a number of factors including species, growth conditions used and developmental stage.
* Average yields will vary depending upon a number of factors including species, growth conditions used and developmental stage.
Storage Conditions and Product Stability All solutions should be kept tightly sealed and stored at room temperature, except for the RNAse which should be stored at -20°C. This kit is stable for 1 year after the date of shipment.
Select Plants and Fungi that can be used with the Plant/Fungi DNA Purification Kits
