The kit offers the unique feature to isolate total RNA including small RNA and DNA from serum and plasma without the need to resort to the cumbersome phenol/chloroform extraction or a time consuming proteinase digest. RNA purified using the kit is ready for applications such as RT-PCR, Northern blotting, poly A+ RNA (mRNA) purification, nuclease protection, and in vitro translation.
Detail
Introduction
The kit offers the unique feature to isolate total RNA including small RNA and DNA from serum and plasma without the need to resort to the cumbersome phenol/chloroform extraction or a time consuming proteinase digest. RNA purified using the kit is ready 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 and miRNA from 0.2-0.6ml serum and plasma
Applications
Purification method
Mini spin column
Purification technology
Silica technology
Process method
Manual (centrifugation or vacuum)
Sample type
Sample amount
Yield
Elution volume
Time per run
Liquid carrying volume per column
Binding yield of column
Principle
HiPure RNA technology simplifies total RNA isolation. The sample material is denatured in Lysis Buffer. The protein is then precipitated by Protein Precipitation Solution and pelleted by centrifugation. After the removal of protein the binding conditions for nucleic acids are adjusted by adding isopropanol. Total nucleic acids are bound to the column. Optionally, DNA can be removed by an on-column DNase digest. The remaining nucleic acids are washed and eluted with minimal amounts of RNase-Free water.
Kit Contents
Contents
R431402
R431403
Purification Times
50 Preps
250 Preps
HiPure RNA Mini Columns
100
2 x 250
2ml Collection Tubes
100
2 x 250
Buffer CFL
6 ml
30 ml
Buffer CFP
1.8 ml
10 ml
Buffer RWC
20 ml
80 ml
Buffer RW2*
20 ml
2 x 50 ml
RNase Free Water
10 ml
60 ml
Storage and Stability
The kit components can be stored at room temperature (15-25°C) and are stable for at least 18 months under these conditions.
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.
This product is a Interleukin-6 Quality Control (two areas) for Milenia QuickLine IL-6 Test (MQL6 1).
This product is only available within the EU!
Method/Platform
control
Range/Assay Sensivity
two areas
Test Principle
The controls are designed only for internal Quality Control of Milenia QuickLine Il-6 Tests. Test results can only be evaluated with Milenia POCScan Reader.
Brief Instructions
Storage
2°C to 8°C
Components
IL-6 Control 1, IL-6 Control 2
Document
2 x 3 flasks
This product is a Interleukin-6 Quality Control (two areas) for Milenia QuickLine IL-6 Test (MQL6 1).
This kit is suitable for extracting total pathogen nucleic acid from a variety of clinical samples such as blood, serum, plasma, swab soaking solution, fluid accumulation and homogenate solution. This kit is designed to remove host cells background nucleic acid and enrich pathogen nucleic acid (including viral/bacterial/fungal DNA/RNA) from the sample. Purified DNA/RNA is ready for downstream applications such as PCR, virus detection, tNGS and other related experiments.
Details
Specifications
Features
Specifications
Main Functions
Extract Pathogen RNA/DNA from 0.5-1.5ml whole blood, plasma, serum, body fluid, homogenate suspension, culture solution, cell suspension, soaking solution or concentrate pathogen solution for tNGS application, remove host background nucleic acid.
Applications
Real Time PCR, biochip analysis, NGS
Products
Pathogen DNA / RNA
Purification method
Polydisperse magnetic beads
Purification technology
Magnetic beads technology
Process method
Manual or automatic
Sample type
whole blood, plasma, serum, body fluid, homogenate suspension, culture solution, cell suspension, soaking solution or concentrate pathogen solution
Sample amount
0.5 – 1.5 ml
Adaptive instrument
Nucleic acid extractor, pipetting workstation
Principle
This product is based on the purification method of high binding magnetic particles. The sample is lysed and digested under the action of lysate and Protease. After adding magnetic particles and binding solution, DNA/RNA will be adsorbed on the surface of magnetic particles, and impurities such as proteins will be removed without adsorption. The adsorbed particles were washed with washing solution to remove proteins and impurities, washed with ethanol to remove salts, and finally DNA/RNA was eluted by Buffer NFW.
Kit Contents
Contents
R667200C
R667202C
Purification Times
24 Preps
96 Preps
2ml Bead Tube (0.4g)
24
96
DNase I (Powder)
10 mg
15 mg
DNase Buffer
5 ml
20 ml
Protease Dissolve Buffer
3 ml
8 ml
Lysis Buffer LBX1
40 ml
180 ml
Buffer TL
5 ml
20 ml
Proteinase K
24 mg
120 mg
MagBind Particles N9
1.2 ml
5 ml
Buffer MLB
30 ml
120 ml
Buffer MW1*
13 ml
110 ml
Buffer MW2*
10 ml
50 ml
Buffer AVE
10 ml
20 ml
Storage and Stability
Proteinase K, DNase I powder and MagPure Particles N9 should be stored at 2–8°C upon arrival. However, short-term storage (up to 8 weeks) at room temperature (15–25°C) does not affect their performance. The remaining kit components can be stored at room temperature (15–25°C) and are stable for at least 18 months under these conditions.
Document
This kit is suitable for extracting total pathogen nucleic acid from a variety of clinical samples such as blood, serum, plasma, swab soaking solution, fluid accumulation and homogenate solution. This kit is designed to remove host cells background nucleic acid and enrich pathogen nucleic acid (including viral/bacterial/fungal DNA/RNA) from the sample. Purified DNA/RNA is ready for downstream applications such as PCR, virus detection, tNGS and other related experiments.
