Application Data
Leeporter Assay Reagent
Leeporter Assay Reagent- 1000 Test
Synonyms
Leeporter Assay Reagent; N/A; Leeporter Assay Reagent- 1000 Test; Leeporter Assay Reagent reagent
Form/Format
1000 test (96-well plate format). Assay buffer contains 0.01% Sodium Azide.
Application Notes
Protocol:
1) Substrate (100X) Reconstitution:
Add 0.5 ml Substrate Reconstitution Solution to lyophilized Substrate vial and dissolve thoroughly by gently pipetting up and down, and/or inverting tubes. The reconstituted 100X substrate may be stored in a dark environment at -20°C for up to one month.
2) Assay Buffer:
Thaw Assay Buffer of 50 ml in room temperature water bath and dissolve thoroughly any precipitates by swirling and/or gentle vortexing until solution becomes clear. Assay Buffer can be aliquoted (e.g. 5-10 ml each) and stored at -20°C for several months.
3) Preparation of complete Assay Solution (for 96-well plate format):
Prepare complete Assay Solution fresh for each use, which should be used within 2 hours. Thaw Assay Buffer and equilibrate to room temperature with swirling and/or gentle vortexing. Calculate how much complete assay solution is needed (Note: 50 ul of complete assay solution is required for each well of 96-well plate). Aliquot proper amount of Assay Buffer in a 15 ml tube and add corresponding amount of reconstituted 100X substrate to make a final 1X substrate assay solution.
4) Luciferase Assay (96-well plate format):
1. Plate you target Leeporter™ Luciferase reporter cells in a white solid-bottom 96-well microplate (Note: The 96-well plate used should be compatible with the luminometer being used.) at 100 ul cells/well, based on the corresponding protocol to your target Leeporter™ cell line (Note: Each target Leeporter™ cell line protocol can be found in its corresponding Data Sheets.). [Total volume per well is 100 uL]
2. Stimulate or treat your target cells based on the corresponding protocol to your target Leeporter™ cell line. [Total volume per well becomes 105~110 uL = 100 ul cells + 5~10 ul treatment]
3. After completion of stimulation/treament of your target cells, equilibrate the 96-well plate containing your target cells being assayed to room temperature for 10 minutes (Note: Do NOT remove or disturb cell culture medium.).
4. Using a multi-channel pipettor, add 50 ul complete Assay Solution directly to each plate well (Note: Do NOT remove cell culture medium when adding the Assay Reagent. So the Assay Reagent of 50 ul should be directly added on top of the cell culture of each well.). [Total volume per well becomes 155~160 uL = 100 ul cells + 5~10 ul treatment + 50 ul complete assay solution]
5. Use automix for 2-3 seconds, and then read the plate in a luminometer within 1-5 minutes. (An example of a luminometer set up: SpectraMaxL (Molecular Devices): target wavelength of 470nm, integration time at 0.5 sec and automix for 2 sec).
1) Substrate (100X) Reconstitution:
Add 0.5 ml Substrate Reconstitution Solution to lyophilized Substrate vial and dissolve thoroughly by gently pipetting up and down, and/or inverting tubes. The reconstituted 100X substrate may be stored in a dark environment at -20°C for up to one month.
2) Assay Buffer:
Thaw Assay Buffer of 50 ml in room temperature water bath and dissolve thoroughly any precipitates by swirling and/or gentle vortexing until solution becomes clear. Assay Buffer can be aliquoted (e.g. 5-10 ml each) and stored at -20°C for several months.
3) Preparation of complete Assay Solution (for 96-well plate format):
Prepare complete Assay Solution fresh for each use, which should be used within 2 hours. Thaw Assay Buffer and equilibrate to room temperature with swirling and/or gentle vortexing. Calculate how much complete assay solution is needed (Note: 50 ul of complete assay solution is required for each well of 96-well plate). Aliquot proper amount of Assay Buffer in a 15 ml tube and add corresponding amount of reconstituted 100X substrate to make a final 1X substrate assay solution.
4) Luciferase Assay (96-well plate format):
1. Plate you target Leeporter™ Luciferase reporter cells in a white solid-bottom 96-well microplate (Note: The 96-well plate used should be compatible with the luminometer being used.) at 100 ul cells/well, based on the corresponding protocol to your target Leeporter™ cell line (Note: Each target Leeporter™ cell line protocol can be found in its corresponding Data Sheets.). [Total volume per well is 100 uL]
2. Stimulate or treat your target cells based on the corresponding protocol to your target Leeporter™ cell line. [Total volume per well becomes 105~110 uL = 100 ul cells + 5~10 ul treatment]
3. After completion of stimulation/treament of your target cells, equilibrate the 96-well plate containing your target cells being assayed to room temperature for 10 minutes (Note: Do NOT remove or disturb cell culture medium.).
4. Using a multi-channel pipettor, add 50 ul complete Assay Solution directly to each plate well (Note: Do NOT remove cell culture medium when adding the Assay Reagent. So the Assay Reagent of 50 ul should be directly added on top of the cell culture of each well.). [Total volume per well becomes 155~160 uL = 100 ul cells + 5~10 ul treatment + 50 ul complete assay solution]
5. Use automix for 2-3 seconds, and then read the plate in a luminometer within 1-5 minutes. (An example of a luminometer set up: SpectraMaxL (Molecular Devices): target wavelength of 470nm, integration time at 0.5 sec and automix for 2 sec).
Components & Storage
- 1 vial of Substrate (100X), lyophilized / -20°C
- 0.5 ml of Substrate Reconstitution Solution / Room Temp. or -20°C
- 50 ml of Assay Buffer / -20°C
- 0.5 ml of Substrate Reconstitution Solution / Room Temp. or -20°C
- 50 ml of Assay Buffer / -20°C
Dry Ice Shipment
Extra charge fee may add to your shipping cost as dry ice is required to ship this product.
Preparation and Storage
Store at -20 degree C
Application Data
Application Data
Related Product Information for Leeporter Assay Reagent reagent
Principle of the Assay: The Leeporter™ Luciferase Assay Reagent was specifically formulated to use with our Leeporter™ (Luciferase Reporter) Cell Lines expressing an optimized intracellular Renilla luciferase, which was designed to produce highly sensitive signal and prolonged signal intensity. As one-step glow assay reagent, the Leeporter™ Luciferase Assay Reagent can be directly added to cell culture plates compatible with the luminometer being used without diluting or transferring culture supernatants or cell lysates to other plates.
Product Categories/Family for Leeporter Assay Reagent reagent
Similar Products
Product Notes
The Leeporter Assay Reagent (Catalog #AAA14875) is a Reagent and is intended for research purposes only. The product is available for immediate purchase. Protocol: 1) Substrate (100X) Reconstitution: Add 0.5 ml Substrate Reconstitution Solution to lyophilized Substrate vial and dissolve thoroughly by gently pipetting up and down, and/or inverting tubes. The reconstituted 100X substrate may be stored in a dark environment at -20°C for up to one month. 2) Assay Buffer: Thaw Assay Buffer of 50 ml in room temperature water bath and dissolve thoroughly any precipitates by swirling and/or gentle vortexing until solution becomes clear. Assay Buffer can be aliquoted (e.g. 5-10 ml each) and stored at -20°C for several months. 3) Preparation of complete Assay Solution (for 96-well plate format): Prepare complete Assay Solution fresh for each use, which should be used within 2 hours. Thaw Assay Buffer and equilibrate to room temperature with swirling and/or gentle vortexing. Calculate how much complete assay solution is needed (Note: 50 ul of complete assay solution is required for each well of 96-well plate). Aliquot proper amount of Assay Buffer in a 15 ml tube and add corresponding amount of reconstituted 100X substrate to make a final 1X substrate assay solution. 4) Luciferase Assay (96-well plate format): 1. Plate you target Leeporter™ Luciferase reporter cells in a white solid-bottom 96-well microplate (Note: The 96-well plate used should be compatible with the luminometer being used.) at 100 ul cells/well, based on the corresponding protocol to your target Leeporter™ cell line (Note: Each target Leeporter™ cell line protocol can be found in its corresponding Data Sheets.). [Total volume per well is 100 uL] 2. Stimulate or treat your target cells based on the corresponding protocol to your target Leeporter™ cell line. [Total volume per well becomes 105~110 uL = 100 ul cells + 5~10 ul treatment] 3. After completion of stimulation/treament of your target cells, equilibrate the 96-well plate containing your target cells being assayed to room temperature for 10 minutes (Note: Do NOT remove or disturb cell culture medium.). 4. Using a multi-channel pipettor, add 50 ul complete Assay Solution directly to each plate well (Note: Do NOT remove cell culture medium when adding the Assay Reagent. So the Assay Reagent of 50 ul should be directly added on top of the cell culture of each well.). [Total volume per well becomes 155~160 uL = 100 ul cells + 5~10 ul treatment + 50 ul complete assay solution] 5. Use automix for 2-3 seconds, and then read the plate in a luminometer within 1-5 minutes. (An example of a luminometer set up: SpectraMaxL (Molecular Devices): target wavelength of 470nm, integration time at 0.5 sec and automix for 2 sec). Researchers should empirically determine the suitability of the Leeporter Assay Reagent for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "Leeporter Assay Reagent, Reagent" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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