Labeling Kit
AminoAllyl Labeling kit for DNA microarrays

Description


This kit has been optimized to be used with 1 to 2 µg of mRNA as starting material, and it can generate 200 to 500 ng of dye labeled cDNA. This kit supports all substrate and slide surface chemistries and provides high yield and efficient incorporation of the dyes. The protocol is easy and the kit is ready-to-use without any buffer or column preparation required. The dye removal reduces background in microarrays hybridizations.
This kit can be used in conjunction with Total RNA extraction kit and with MiniAmp amplification kit.

Reference Designation
AFK Indirect Amino Allyl Fluorescent Labeling Kit (20 labeling reactions)

Specification


Needed Equipment

• Dye such as Amersham CyDye™ Set Cat # RPN5661, Alexa Fluor® 555 and Alexa Fluor® 647, Pierce DyLight 547 and 647 Fluors etc
• Ethanol 100%
• Ethanol 80%
• Ice for incubation
• PCR tubes
• Thermal cycler
• Microfuge capable of 10,000 rpm or more
• Micropipettors
• Vortex mixer

Kit Content

Articles Conditionnement Stockage
 Random Primer  22 µl in 0.5ml tube  -20°C Freezer
 dTVN Primer  22 µl in 0.5ml tube  -20°C Freezer
 Labeling Enzyme Mix  33 µl in 0.5ml tube  -20°C Freezer
 Labeling Reaction Buffer  44 µl in 0.5ml tube  -20°C Freezer
 AA-dNTP Mix  88 µl in 0.5ml tube  -20°C Freezer
 DTT  22 µl in 0.5ml tube  -20°C Freezer
 cDNA Synthesis Stop Solution  220 µl in 0.5ml tube  -20°C Freezer
 Denaturing Solution  220 µl in 0.5ml tube  -20°C Freezer
 Neutralization Solution  220 µl in 0.5ml tube  -20°C Freezer
 Dye Binding Buffer  500 µl in 2ml tube  -20°C Freezer
 Dye Binding Stop Solution  110 µl in 0.5ml tube  -20°C Freezer
 DNA Binding Buffer  6.6 ml in 8ml bottle  Room Temp
 DNA Wash Buffer Concentrate  1.5 ml in 8ml bottle  Room Temp
 DNA Micro Column  30  Room Temp
 Elution Tube (1.5ml)  30  Room Temp
 2 ml Wash Tube  30  Room Temp
 Nuclease Free Water  1.5ml in 2ml tube  -20°C Freezer

Short Protocol

1. Perform cDNA Synthesis from mRNA .
2. Perform hydrolysis of RNA.
3. Purify the Amino allyl labeled cDNA
4. Dye binding with Amino allyl labeled cDNA.
5. Purify the dye labeled cDNA.

Please contact This e-mail address is being protected from spambots. You need JavaScript enabled to view it to get further information about the complete protocol.


Protein Labeling kit for protein microarrays

Description


This labelling kit for proteins includes fluorescent dyes, reagents and purification columns for a high quality and yield direct labeling of proteins for protein microarrays. The simple, easy-to-use and flexible protocol can be used with antibodies, secondary IgG, enzymes and other proteins.
The amine dyes provided with this kit are excellent alternative to the Cy3 and Cy5 dyes used with most popular microarrays scanners. Their absorption and emission wavelength are suitable with the scanner filters, with no need to purchase new filter sets.
 
Reference Designation
Please contact us 2 color fluorescent Labeling Kit for proteins

Specification


Needed Equipement


• Antibodies microarrays: Protein microarrays, PlasmaScan antibodies microarrays, Tissue microarrays etc.
• Microarrays instruments: TrayMix hybridization stations, Ozone Free Box etc.
• Reaction tubes
• Micropipettors
• Vortex mixer
• Microcentrifuge

Kit
Content


Please contact This e-mail address is being protected from spambots. You need JavaScript enabled to view it for further information about thid kit.

Short protocol for 1 mg protein

1. Dissolve protein 1 mg per 100 µL in PBS in a reaction tube
2 Add 20 µL of reaction solution A to the protein reaction tube.
3. Prepare the reactive Arrayit Green540 and Arrayit Red640 stock just prior to starting the reaction.
Add 25 µL of solution B to 1 tube of dye and pipette up and down to mix and dissolve.
4. Combine 10 µL of reactive dye solution to the protein reaction tube and vortex gently to mix.
Dye solution should be stored at 4C and can be used once more within 1 week.
5. Let reaction run for 1 hour at room temperature protected from light.
6. While reaction runs, prepare two purification columns.
7. Stop dye labeling reaction using buffer D (30 minutes).
8. Purify reaction by transferring half of the protein reaction tube contents to two purification columns and spin at 750 x g for 2 minutes to collect dye conjugated proteins.