Propargyl-PEG24-amine is a linker with 24 PEG units. The amine group is reactive with carboxylic acids, activated NHS esters, carbonyls (ketone, aldehyde) etc. The propargyl group is reactive with azide compounds under the catalyzation of copper. This linker has pretty good water-solubility due to its long PEG chain. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

Propargyl-PEG24-amine is a linker with 24 PEG units. The amine group is reactive with carboxylic acids, activated NHS esters, carbonyls (ketone, aldehyde) etc. The propargyl group is reactive with azide compounds under the catalyzation of copper. This linker has pretty good water-solubility due to its long PEG chain. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

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.

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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).

‍[4] Membrane alterations (phosphatidylserine flip).

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.

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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.

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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.

The APOPercentage™ Apoptosis kit is a dye-based, colorimetric assay for detection and measurement of apoptosis (programmed cell death) during in-vitro cell culture.

Permagen’s  50 mL Centrifuge Tube Magnetic rack is designed for magnetic bead separations from up to six, 50 mL Centrifuge tubes

Accommodates any common 50 mL Centrifuge Tubes

Beads will be pulled to back wall allowing easy aspiration and tip tracking down the front wall of the tubes without disturbing bead pellet

Features include solid aluminum alloy design with hard coat finish for years of trouble-free use, rubber feet to help prevent slipping on work bench, less tippy than common plastic products, and fast separations using any magnetic beads

SKU #

MSR6X50

Features

• Aluminum alloy design for years of trouble-free use
• Fast magnetic bead separations
• Held to tighter tolerances for more consistent results
• Pulls beads to side of wells for easy aspiration without disturbing bead pellet 
• Rubber feet to help prevent slippage
• Stable design
• Four magnets per tube 

Compatibility

• Up to six 50 mL Centrifuge Tubes
• Any Magnetic Beads

Volumes

Maximum Volume – 50 mL

Minimum Volume – 10 mL

Permagen’s 50 mL Centrifuge Tube Magnetic rack is designed for magnetic bead separations from up to six, 50 mL Centrifuge tubes

Description 

ExcelTaq™ Klen-Taq DNA Polymerase is a specially blended enzyme mix containing KlenTaq-1 DNA polymerase (a 5’-exo-minus, N-terminal deletion of Taq DNA polymerase) and a small amount of a proofreading DNA polymerase. This unique blending helps to improve the fidelity, yield and processivity of the resultant PCR process. The Klen-Taq is also highly robust, showing high tolerance of varying concentrations of Mg²⁺; it is highly thermostable and has four times the fidelity compared to Taq DNA polymerase. The ExcelTaq™ Klen-Taq DNA Polymerase is ideal for DNA amplifications 0.5-5 kb in length on genomic DNA, and up to 10 kb on less complex templates.

Features

• 5’→3′ DNA polymerase activity
• 3’→5′ exonuclease activity (proofreading)
• 4× fidelity as compared to Taq DNA polymerase
• Thermo-stable: up to 98°C during PCR denaturing step
• Robust PCR performance, resistance to variance in PCR conditions 

Storage

-20°C for 24 months 

ExcelTaq™ Klen-Taq DNA Polymerase is a specially blended enzyme mix containing KlenTaq-1 DNA polymerase (a 5’-exo-minus, N-terminal deletion of Taq DNA polymerase) and a small amount of a proofreading DNA polymerase. This unique blending helps to improve the fidelity, yield and processivity of the resultant PCR process. The Klen-Taq is also highly robust, showing high tolerance of varying concentrations of Mg2+; it is highly thermostable and has four times the fidelity compared to Taq DNA polymerase. The ExcelTaq™ Klen-Taq DNA Polymerase is ideal for DNA amplifications 0.5-5 kb in length on genomic DNA, and up to 10 kb on less complex templates.

Introduction

This product is suitable for rapid extraction of DNA from FFPE sample. This kit uses two combination methods. High-salt Bind is conducive to remove pigments or polysaccharides from complex FFPE samples, so as to improve the purity of nucleic acid and avoid blocking aligent 2100. Alcohol mediated adsorption is conducive to improve the nucleic acid yield of high-yield samples.

Details

Specifications

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 yield – most optimal process, recovery up to 90%
• Economy – less than 50% of the price of Qiagen and other imported products
• High purity – OD 260/280 : 1.7-1.9, OD 260/230 : 1.5-2.0

Kit Contents

Storage and Stability

Proteinase K, RNase A and MagPure Particles N 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 at room temperature (15-25°C) and are stable for at least 18 months under these conditions.

This product is suitable for rapid extraction of DNA from FFPE sample. This kit uses two combination methods. High-salt Bind is conducive to remove pigments or polysaccharides from complex FFPE samples, so as to improve the purity of nucleic acid and avoid blocking aligent 2100. Alcohol mediated adsorption is conducive to improve the nucleic acid yield of high-yield samples.