1.Usages: For fungal culture and sterile inspection of drugs and biological products.
Principle: Peptone provide carbon and nitrogen sources; yeast extract powder provides B vitamins; glucose provide energy; potassium hydrogen phosphate as buffer; magnesium sulfate to provide essential trace elements.
Storage: Keep container tightly closed, store in a cool, dry place, away from bright light. Storage period of 3 years.
Other Products
m-PEG4-DBCO
Product Info
Document
Product Info
m-PEG4-DBCO is a click chemistry reagents which can enable so called copper-free Click Chemistry with azide to form a stable triazole. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
Document
m-PEG4-DBCO is a click chemistry reagents which can enable so called copper-free Click Chemistry with azide to form a stable triazole. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
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.
The Kit provides fast purification of high-quality RNA from plants, cell, tissues, and yeast using silica-membrane spin columns with a binding capacity of 100μg RNA. There is no need for phenol/chloroform extractions and time-consuming steps such as CsCl gradient ultracentrifugation or precipitation with isopropanol or LiCl. RNA purified using the RaPure Total RNA Purification System can be used for applications such as RT-PCR, Northern blotting, poly A+ RNA (mRNA) purification, nuclease protection, and in vitro translation.
Details
Specifications
Features
Specifications
Main Functions
Isolation total RNA from <150 mg simple plant sample without chloroform
Applications
RT-PCR, qPCR, Northern hybridization, second generation sequencing, nucleic acid protection, in vitro translation
Purification method
Mini spin column
Purification technology
Silica technology, DNA filtration technology
Process method
Manual (centrifugation or vacuum)
Sample type
Economic crops
Sample amount
≤150 mg
Elution volume
≥30μl
Time per run
≤25 minutes
Liquid carrying volume per column
800µl
Binding yield of column
100µg
Principle
The Kit isolates total RNA from up to 150mg plant tissue. A short workflow enables RNA isolation with genomic DNA removal in less than 25 min. Samples are first lysed and homogenized. The lysate is passed through a DNA Mini column, ethanolis added to the flow-through, and the sample is applied to an RNA column. RNA binds to the membrane and contaminants are washed away. High-quality RNA is eluted in as little as 30µl water using the Kit.
Advantages
Efficient removal of DNA – unique genomic DNA removal column without DNase treatment
High quality – high purity total RNA can be directly used in various sensitive downstream applications
Fast – several samples can be extracted in 25 minutes by column method
Safe – no phenol chloroform extraction required
Sensitive – RNA can be recovered at the level of PG
Broad spectrum – various types of plant samples can be processed by diversity of operating procedures
Kit Contents
Contents
R415102
D415103
Purification Times
50 Preps
250 Preps
HiPure DNA Mini Column II
50
250
HiPure RNA Mini Columns
50
250
2ml Collection Tubes
100
2 x 250
Buffer RLC
50 ml
200 ml
Buffer PRC1
50 ml
200 ml
Buffer RW1
50 ml
200 ml
Buffer RW2*
20 ml
2 x 50 ml
RNase Free Water
10 ml
30 ml
Storage and Stability
The Kit can be stored dry at room temperature (15-25°C) and are stable for at least 18 months under these conditions. Make sure that all buffers are at room temperature when used. During shipment, crystals or precipitation may form inthe Buffer RLC/PRC1. Dissolve by warming buffer to 37°C.
Purchase Guide
1. When dealing with woody or uncommon samples, R4150 is recommended first. R4150 contains two polysaccharide/polyphenol lysis buffer, which is the most universal product.
2. R4151 is recommended for handling common economic crop samples for the first time. Strong lysis solution can be used to process easy-extraction samples. The amount of corn or rice leaves samples can reach up to 300mg.
3. R4165 adopts CTAB/chloroform method, which can also handle a large number of difficult-to-extraction plants, but requires contact with chloroform substitutes, which is less safe than other kits. This kit uses DNase Ⅰ to remove DNA, which is also a good choice for extracting polysaccharide/polyphenol-rich plant samples.
4. R4014 is recommended for fruit/starch plant samples, which uses improved trizol pre-treatment, single column operation and is more economical.
Select the right purification kit to get impactful results：
The Kit provides fast purification of high-quality RNA from plants, cell, tissues, and yeast using silica-membrane spin columns with a binding capacity of 100μg RNA. There is no need for phenol/chloroform extractions and time-consuming steps such as CsCl gradient ultracentrifugation or precipitation with isopropanol or LiCl. RNA purified using the RaPure Total RNA Purification System can be used for applications such as RT-PCR, Northern blotting, poly A+ RNA (mRNA) purification, nuclease protection, and in vitro translation.
