【IT1000】EzRNA™ T7 High Yield RNA Synthesis Kit, 50 RXN
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The EzRNA™ T7 High Yield RNA Synthesis Kit is a user-friendly product for enzymatic RNA production. The enzyme mix contains adequate amount of T7 RNA polymerase, pyrophosphatase, and RNase inhibitors for in vitro transcription (IVT). Along with 10X Transcription Buffer and NTP Premix, users can swiftly assemble IVT reactions without compromising RNA yield. The EzRNA™ T7 High Yield RNA Synthesis Kit allows for the attainment of approximately up to 150 µg RNA yield within 2 hours at 37°C.
Detail
Description
The EzRNA™ T7 High Yield RNA Synthesis Kit is a user-friendly product for enzymatic RNA production. The enzyme mix contains adequate amount of T7 RNA polymerase, pyrophosphatase, and RNase inhibitors for in vitro transcription (IVT). Along with 10X Transcription Buffer and NTP Premix, users can swiftly assemble IVT reactions without compromising RNA yield. The EzRNA™ T7 High Yield RNA Synthesis Kit allows for the attainment of approximately up to 150 µg RNA yield within 2 hours at 37°C.
Features
High yield
Versatile- suitable for short and long transcripts
NTP premixed- Minimal pipetting and setup time
Compatible with CleanCap® Reagent AG
Lithium chloride included for RNA purification
Application
Generation of RNA from T7 promoter-driven DNA sequences
Suitable for subsequent cap-0 and cap-1 modification
Storage
-20°C for 12 months
Other Products
DBCO-NHCO-PEG4-amine, TFA salt
Product Info
Document
Product Info
DBCO-NHCO-PEG4-amine is a carboxyl-reactive building block with extended PEG spacer arm. The hydrophilic PEG spacer arm improves water solubility. In the presence of of activators (e.g. EDC, or HATU), this reagent can be used to derivatize carboxyl groups or activated esters (e.g. The NHS ester) through a stable amide bond. DBCO is commonly used for copper-free Click Chemistry reactions. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
Document
DBCO-NHCO-PEG4-amine is a carboxyl-reactive building block with extended PEG spacer arm. The hydrophilic PEG spacer arm improves water solubility. In the presence of of activators (e.g. EDC, or HATU), this reagent can be used to derivatize carboxyl groups or activated esters (e.g. The NHS ester) through a stable amide bond. DBCO is commonly used for copper-free Click Chemistry reactions. 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.
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.
