The product is made of medical grade USP CLASS VI polymer polystyrene
The product is made under a 100,00- class dust-free manufacturing site
Two kinds of product line up are providing.
For adherent cell culture: Initial adherence and proliferative property of cells via hydrophilic surface treatment.
For suspension cell culture: The surface is resistant to cell adherence, which minimizes damage or loss of cell.
Gamma radiation sterilization
Non-Pyrogenic, DNase/Rnase free.
Other Products
P1006 HiPure Plasmid Plus 96 Kit
Product Info
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Product Info
Introduction
The HiPure Plasmid DNA Plus 96 Kit enables up to 96 minipreps to be performed simultaneously in less than 45 minutes on the Vacuum Manifold (Qiavac 96). This kit provides a fast, simple,and cost-effective plasmid DNA high-throughput method for routine molecular biology laboratory applications. HiPure Kits use silica membrane technology to eliminate the cumbersome steps associated with loose resins or slurries.Plasmid DNA purified with Kits is immediately ready for use. Phenol extraction and ethanol precipitation are not required, and high quality plasmid DNA is eluted in a small volume of Tris buffer or water.
Details
Specifications
Features
Specifications
Main Functions
Isolation up to 20µg plasmid DNA from 1.5ml bacterial culture using 96 well bind plate and 96 filterplate
Applications
Enzyme digestion, sequencing, PCR, labeling, etc.
Purification method
96 well plate
Purification technology
Silica technology
Process method
Manual (centrifugation or vacuum)
Sample type
Conventional plasmid, plasmid less than 30KB
Sample amount
1-1.5ml（x96）
Yield
1-15µg/1ml
Elution volume
≥70μl
Time per run
≤60 minutes
Liquid carrying volume per column
800µl
Binding yield of column
70µg
Principle
The HiPure Plasmid procedure is based on alkaline lysis of bacterial cells followed by adsorption of DNA onto silica in the presence of high salt. The unique silica membrane used in the kit completely replaces glass or silica slurries for plasmid DNA minipreps. The procedure consists of 3 basic steps: Preparation and clearing of a bacterial lysate by alkaline method，then transfer the supernatant to column to bind DNA. After washing proteins and other impurities, nucleic acid was finally eluted with low-salt buffer (10mm Tris, pH9.0, 0.5mm EDTA).
Advantages
High purity – purified plasmid can be directly used in sequencing, enzyme digestion and PCR, etc.
Fast – it takes only 60 minutes to complete the isolation
High yield – up to 1mg plasmid can be binded in one column
Economy – high cost performance
Kit Contents
Contents
P100601
P100602
P100603
Purification Times
1 x 96 Preps
4 x 96 Preps
20 x 96 Preps
RNase A
5 mg
20 mg
100 mg
Buffer P1
30 ml
120 ml
600 ml
Buffer P2
30 ml
120 ml
600 ml
Buffer P3
40 ml
180 ml
800 ml
Buffer PW1
100 ml
500 ml
2 x 1000 ml
Buffer PW2
50 ml
2 x 100 ml
4 x 200 ml
Elution Buffer
150 ml
60 ml
300 ml
Lysate Clear Plate
1
4
20
HiPure DNA Plate
1
4
20
1.6 ml Collection Plate
1
4
20
0.5ml Elute Plate
1
4
20
Storage and Stability
The kit components can be stored dry at room temperature (15-25°C) and are stable for at least 18 months under these conditions. If any precipitates form in the buffers, warm at 37℃ to dissolve. After addition of RNase A，Buffer P1 is stable for 6 months when stored at 2-8°C.
The HiPure Plasmid DNA Plus 96 Kit enables up to 96 minipreps to be performed simultaneously in less than 45 minutes on the Vacuum Manifold (Qiavac 96). This kit provides a fast, simple,and cost-effective plasmid DNA high-throughput method for routine molecular biology laboratory applications. HiPure Kits use silica membrane technology to eliminate the cumbersome steps associated with loose resins or slurries.Plasmid DNA purified with Kits is immediately ready for use. Phenol extraction and ethanol precipitation are not required, and high quality plasmid DNA is eluted in a small volume of Tris buffer or water.
Round well with ‘V’ bottom for maximum liquid extraction
High uniformity from well to well
Raised well rims for reliable closing with heat sealing.
Easy and reliable stacking
Good centrifugation stability up to 6,000 × g for faster protocols and improved sample quality
Manufactured under DNase/ RNase free environment without slip agents, plasticizers or biocides – Materials which could have a negative effect on bioassays
Pure, virgin polypropylene guarantees good extractible performance, high resistance to chemicals, good mechanical stress and working with temperature extremes
Autoclavable (121 °C, 20 min)
Document
Working volume of 0.36ml Round well with ‘V’ bottom for maximum liquid extraction High uniformity from well to well Raised well rims for reliable closing with heat sealing. Easy and reliable stacking Good centrifugation stability up to 6,000 × g for faster protocols and improved sample quality Manufactured under DNase/ RNase free environment without slip agents, plasticizers or biocides – Materials which could have a negative effect on bioassays Pure, virgin polypropylene guarantees good extractible performance, high resistance to chemicals, good mechanical stress and working with temperature extremes Autoclavable (121 °C, 20 min)
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.
