
20 bp DNA Ladder on 3.6% of agarose gel, 8% and 12% of acrylamide gel.
• For sizing and quantification of double strand DNA fragments.
• Composed of eight bands as shown on right.
• Premixed with 6X DNA loading buffer for direct gel loading.
• For sizing and quantification of double strand DNA fragments.
• Composed of eight bands as shown on right.
• Premixed with 6X DNA loading buffer for direct gel loading.
20 bp DNA Ladder on 3.6% of agarose gel, 8% and 12% of acrylamide gel.
• For sizing and quantification of double strand DNA fragments.
• Composed of eight bands as shown on right.
• Premixed with 6X DNA loading buffer for direct gel loading.
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.
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).
[4] Membrane alterations (phosphatidylserine flip).
Biocolor’s APOPercentage assay is based on the latter. Further information can be found under the ‘Mode of Action’ Tab.
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.
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.
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.
A single cell (via image analysis method)
Colorimetric (550nm) (Endpoint) or Image Analysis based
Sufficient for 4×24 well plates or 6×96 well plates
Adherent mammalian cells (in-vitro)
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.
The APOPercentage™ Apoptosis kit is a dye-based, colorimetric assay for detection and measurement of apoptosis (programmed cell death) during in-vitro cell culture.
Norgen’s EXTRAClean Urine Exosome Purification and RNA Isolation Mini Kit constitutes an all-in-one system for the purification of exosomes and the subsequent isolation of RNA from different urine sample volumes ranging from 250 μL to 1 mL. The purification is based on spin column chromatography that employs Norgen’s proprietary resin. The EXTRAClean columns undergo stringent processing and rigorous quality control measures to minimize contamination traces, ensuring optimal results for sensitive applications such as NGS. The kit is designed to isolate all sizes of RNA, including microRNA. The kit provides a clear advantage over other available kits in that it does not require any special instrumentation, protein precipitation reagents, extension tubes, phenol/chloroform or protease treatments. Moreover, the kit allows the user to elute into a flexible elution volume ranging from 50 μL to 100 μL. The purified RNA is free from any protein-bound circulating RNA and of the highest integrity. The purified RNA can be used in a number of downstream applications including real time PCR, reverse transcription PCR, NGS application, Northern blotting, RNase protection and primer extension, and expression array assays.
Figure 1 / 9
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Minimum Urine Input | 250 μL |
Maximum Urine Input | 1 mL |
Size of Exosomes Purified | 40 nm – 150 nm |
Size of RNA Purified | All sizes, including miRNA and small RNA (< 200 nt) |
Elution Volume | 50-100 μL |
Time to Complete 10 Purifications | 35-40 minutes |
Average Yields | Variable depending on specimen |
Storage Conditions and Product Stability
All buffers should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment. It is recommended to warm Lysis Buffer A for 20 minutes at 60°C if any salt precipitation is observed.
Magen’s HiPure columns are prepared by high quality glass fiber filter membrane as raw materials through membrane cutting, membrane release, ring release, ring pressing, gland, weighing and other processes. HiPure nucleic acid adsorption columns have the characteristics of long-term stability and high binding capacity. Experiments show that the highest binding capacity and binding efficiency of HiPure nucleic acid adsorption columns are basically unchanged when stored at room temperature for 4 years.
The series of nucleic acid columns produced by Magen Biotech are based on carefully selected imported glass fiber membranes (GF/B, GF/D, GF/F). Columns production processes such as polypropylene injection molding materials, injection molding process, and downstream membrane packing and compression rings are strictly controlled. This is to ensure that the column has extremely high adsorption capacity and long-term stability. Compared with conventional products on the market, Magen’s columns are with varieties, and binding rate will not change when stored at room temperature for 4 years.
Specifications
Features | Specifications |
Recommended application | Plasmid Mini Preparation,gDNA/ RNA Extraction, DNA/RNA Clean Up |
Preservation conditions | Room temperature |
Stability | Up to 4 years |
Filter membrane | High quality glass fiber filter GF/B, 4 layers |
Membrane aperture | 1.0μm |
Maximum binding yield of plasmid | 35 μg |
Maximum yield of alcohol mediated Binding | 200 μg |
Single liquid carrying capacity of column | 800 μl |
Minimum elution volume | 50 μl |
Withstand centrifugal force | 16,000 x g |
Centrifuge | Small high speed centrifuge (2ml) |
Adsorption Mechanism
Based on the negatively charged DNA skeleton, it has a high affinity for positively charged glass fibers. In high salt and ethanol solutions, DNA/RNA binds to glass fiber and interacts with hydrophilic matrix on silica through hydrogen bond. DNA/RNA is tightly bound. All pollutants can be removed by washing solution. At high salt concentration, nucleic acids selectively bind to silicagel membrane, while other pollutants, mainly proteins, are removed by membrane washing.
Ordering information
CAT.No. | Product Name | Package |
C13110 | HiPure DNA Mini Column II (4 x GF/B)with 2ml Collection Tubes | 1000/Bag |
Item No. | Product Name | Membrane type/number of layers | Collection tubes | Plasmid DNA binding capacity (Physical adsorption) | gDNA/RNA binding capacity (Alcohol-mediated adsorption) | Minimum Elution volume | Liquid volume capacity |
C13010 | HiPure DNA Nano Column | 2 layers GF/F | 2ml without cap | 5μg | 20μg | 10μl | 700μl |
C13011 | HiPure DNA Micro Column | 3 layers GF/F | 2ml without cap | 10μg | 50μg | 15μl | 700μl |
C13100 | HiPure DNA Mini Column I | 2 layers GF/B | 2ml without cap | 15μg | 100μg | 30μl | 700μl |
C13110 | HiPure DNA Mini Column II | 4 layers GF/B | 2ml without cap | 35μg | 200μg | 50μl | 800μl |
C13111 | HiPure RNA Mini Column | 3 layers GF/B | 2ml without cap | 30μg | 200μg | 30μl | 800μl |
C13112 | HiPure Viral Mini Column | 3 layers GF/F | 2ml without cap | 30μg | 200μg | 30μl | 800μl |
C13113 | HiPure CFDNA Mini Column | 3 layers GF/F,1 layer GF/B | 2ml without cap | 30μg | 200μg | 30μl | 800μl |
C13120 | HiPure DNA Midi Column | 4 layers GF/B | 15ml Centrifuge tube | 125μg | 1mg | 500μl | 4ml |
C13121 | HiPure DNA Midi Column III | 8 layers GF/B | 15ml Centrifuge tube | 250μg | 1mg | 500μl | 4ml |
C13122 | HiPure DNA Maxi Column | 4 layers GF/B | 50ml Centrifuge tube | 500μg | 5mg | 1000μl | 20ml |
C13123 | HiPure DNA Maxi Column III | 8 layers GF/B | 50ml Centrifuge tube | 1mg | 5mg | 1000μl | 20ml |
C13124 | HiPure DNA Maxi Column C | 8 layers GF/B | 50ml high speed Centrifuge tube | 1mg | 5mg | 700μl | 12ml |
C13130 | HiPure DNA Plate | 2 layers GF/F | 1.6ml Plate | 30μg | 100μg | 80μl | 900μl |
C13131 | HiPure gDNA Plate | 2 layers GF/B | 1.6ml Plate | 30μg | 100μg | 80μl | 900μl |
Note: GF/B pore size is for 1.0μM glass fiber membrane; GF/F pore size is for 0.7μm glass fiber membrane.
Magen’s HiPure columns are prepared by high quality glass fiber filter membrane as raw materials through membrane cutting, membrane release, ring release, ring pressing, gland, weighing and other processes. HiPure nucleic acid adsorption columns have the characteristics of long-term stability and high binding capacity. Experiments show that the highest binding capacity and binding efficiency of HiPure nucleic acid adsorption columns are basically unchanged when stored at room temperature for 4 years.
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Email : hej@a3p-scientific.com
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