| Description |
| The Chemiluminescent Array Detection Kit contains all of the reagents
required to process GEArrays® for the detection of biotinylated probes or
targets hybridized to the array. Each kit contains enough reagents to
process twelve of any of the GEArray® nylon membranes (cDNA Q and S
Series GEArrays® and Oligo GEArrays® and to perform up to four quality
control checks of Probe or cRNA Target Synthesis. Instructions for the use
of this kit are included in your GEArray User Manual. A brief protocol for
experienced array users is also provided below.
Need more information about GEArray products? please visit Microarray Home or send Email to Technical Support |
| Kit Contents /
Packing List / Storage Conditions |
| Please check the kit components immediately after you receive
this package. SABiosciences is not responsible for any missing items not
reported within two (2) business days upon receipt.
Storage Conditions: The following kit is shipped at ambient
temperature. For long-term storage, keep entire kit at 4 ºC
| Chemiluminescent Array Detection Kit
|
| Component
|
| GEAblocking
Solution Q |
| AP-Streptavidin |
| 5X Buffer F (5X
Washing Buffer) |
| Buffer G (AP
Assay Buffer) |
| CDP-Star
Substrate |
|
| Brief Protocol |
Note: All of the following detection steps are performed at room
temperature. Be sure to allow your hybridization oven and
cylinders and our hybridization tube to cool before continuing.
Note: GEAblocking Solution Q and 5X Buffer F may cloud during
storage at 4 ºC. Warm the solutions to 37 ºC and invert the bottles
several times to allow any precipitate to completely dissolve. Allow the
solutions to sit at room temperature until needed.
- Blocking the Array:
Discard the last wash and add 2 ml GEAblocking Solution Q.
Incubate for 40 min with continuous agitation at 20 to 30 rpm.
- Binding of alkaline phosphatase-conjugated streptavidin (AP):
Prepare Binding Buffer:
Dilute 5X Buffer F five-fold to prepare excess 1X Buffer F. Dilute AP
1:8,000 into 1X Buffer F to obtain your binding buffer. We suggest
dispensing volumes of AP no smaller than 2 µl. You will also need 16
ml of 1X Buffer F per tube for washing (3).
Discard the GEAblocking Solution Q from the tube. Add 2 ml Binding
Buffer, and incubate for 10 min with continuous but gentle (5-10 rpm)
agitation.
- Washing:
Wash the membrane four times with 4 ml 1X Buffer F for 5 min with
gentle agitation. Vortex the tube gently after each addition of fresh
1X Buffer F. Rinse or wash twice with 3 ml Buffer G.
- Detection:
Add 1.0 ml CDP-Star chemiluminescent substrate to the hybridization
tube. Incubate at room temperature for 2-5 min.
Note: It is very important to have the membrane covered evenly with
the substrate.
Blot the membrane on a piece of filter paper to remove excess CDP-Star
Solution.
Do not let the membrane completely dry out.
The membrane should be saturated and translucent without any solution
dripping from it.
Place the membrane into a plastic sheet protector or into a small
plastic zip-lock bag and smooth out any bubbles.
|
