Brevetoxins (PbTx) are a class of cyclic polyether compounds produced by certain algae such as Karenia brevis. K. brevis can produce Brevetoxins in large quantities during an algae bloom which then pose a major health threat and are important to monitor and mitigate. Brevetoxins are the causative agent of Neurotoxic Shellfish Poisoning (NSP).
FDA and EPA safety levels in regulations and guidance – 0.8 mg/kg for clams, mussels, oysters, and whole and roe-on scallops, fresh, frozen, or canned. – National Shellfish Sanitation Program Guide for the Control of Molluscan Shellfish.
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IVD3101 MagPure DNA Micro Kit
Product Info
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Product Info
Introduction
This product is suitable for rapid extraction of DNA from tissue, cells, blood, saliva, swabs, blood spots, semen and other clinical samples. DNA can be used directly for PCR, quantitative PCR, Southern Blot, test of virus DNA and so on.
Details
Specifications
Features
Specifications
Main Functions
Isolation total DNA from 1-100ul blood, FFPE, tissue and other samples
Applications
Second generation sequencing, PCR, real timePCR, etc.
Purification method
Monodisperse magnetic beads
Purification technology
Magnetic beads technology
Process method
Manual or automatic
Sample type
Anticoagulant blood, concentrated blood, buffy coat, lymphocytes and cultured cells
Sample amount
Body fluid : 10 – 200μl, Tissue : ≤10mg
Yield
10ng – 15μg
Elution volume
80 -100μl
Time per run
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. DNA is released into the lysate. After adding magnetic particles and binding solution, DNA 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 was eluted by Elution Buffer.
Advantages
High binding force – suitable for handling DNA rich samples, such as whole blood, buffy coat, concentrated blood, etc.
High purity – carboxyl magnetic beads with weak surface adsorption, getting higher purity
General – can be used for various clinical samples
Kit Contents
Contents
IVD3101
Purification Times
200
MagBind Particles
4.5 ml
Proteinase K
90 mg
Protease Dissolve Buffer
10 ml
Buffer ATL
60 ml
Buffer AL
60 ml
Buffer BD*
20 ml
Buffer BXW1*
110 ml
Elution Buffer
30 ml
Storage and Stability
Proteinase K, MagBind Particles should be stored at 2–8°C upon arrival. However, short-termstorage (up to 18 weeks) at room temperature (15–25°C) does not affect their performance. The remainingkit components can be stored dry at room temperature (15–25°C) and are stable for at least 18 months under these conditions.
Document
This product is suitable for rapid extraction of DNA from tissue, cells, blood, saliva, swabs, blood spots, semen and other clinical samples. DNA can be used directly for PCR, quantitative PCR, Southern Blot, test of virus DNA and so on.
HiPure Soil RNA Kit is suitable for extracting high-purity microbial total RNA from soil samples. The kit adopts silica gel column purification technology and original humic acid adsorbent technology. It is suitable for extracting high-yield and high-purity total RNA from various soil samples, such as forest soil, grassland soil, mining soil, sediment and so on. The obtained RNA can be directly used in RT-PCR, Northern blot, poly-A + purification, nucleic acid protection and in vitro translation.
Details
Specifications
Features
Specifications
Main Functions
Isolation total RNA from 500 mg soil sample
Applications
RT-PCR, Northern blot, poly-A + purification, nucleic acid protection and in vitro translation
Purification method
Mini spin column
Purification technology
Silica technology
Process method
Manual (centrifugation or vacuum)
Sample type
Forest soil, grassland soil, mining area soil, sediment and other samples
Sample amount
500 mg
Elution volume
≥30μl
Time per run
≤60 minutes
Liquid carrying volume per column
800µl
Binding yield of column
100µg
Principle
Hipure silica gel column is based on glass fiber filter membrane with high binding force. Under the condition of high concentration of ionizing agent (such as guanidine hydrochloride or guanidine isothiocyanate), the filter membrane can adsorb nucleic acid through hydrogen bond and electrostatic, while protein and other impurities are not adsorbed and removed. The filter membrane adsorbed with nucleic acid is washed to remove the residual protein and salt. Finally, the nucleic acid adsorbed on the filter membrane can be washed out with low salt buffer (such as buffer TE) or water. The obtained nucleic acid has high purity and can be directly used in various downstream experiments.
Advantages
High quality – high purity total RNA can be directly used in various sensitive downstream applications
Fast – extraction of several samples can be completed in 60 minutes by column purification method
Safe – no phenol chloroform extraction required
Sensitive – RNA can be purified at the level of PG
Kit Contents
Contents
R418302
R418303
Purification Times
50 Preps
250 Preps
HiPure RNA Micro Columns
50
250
2ml Collection Tubes
100
500
gDNA Filter Column
50
250
2ml Beads Tubes
50
250
Buffer SOL
30 ml
150 ml
Buffer SDS
4 ml
15 ml
Buffer PHC
30 ml
150 ml
Buffer GDP
40 ml
150 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
Buffer PHC should be stored at 2–8°C upon arrival. However, short-term storage (up to 12 weeks) at room temperature (15–25°C) does not affect their performance. The remaining kit components can be stored dry at room temperature (15–25°C) and are stable for at least 18 months under theseconditions.The entire kit can be stored at 2–8°C, but in this case buffers should be redissolvedbefore use. Make sure that all buffers are at room temperature when used.
Experiment Data
Document
HiPure Soil RNA Kit is suitable for extracting high-purity microbial total RNA from soil samples. The kit adopts silica gel column purification technology and original humic acid adsorbent technology. It is suitable for extracting high-yield and high-purity total RNA from various soil samples, such as forest soil, grassland soil, mining soil, sediment and so on. The obtained RNA can be directly used in RT-PCR, Northern blot, poly-A + purification, nucleic acid protection and in vitro translation.
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.
