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AlphaLISA SureFire Ultra Human Total Progesterone Receptor Detection Kit, 10,000 Assay Points
AlphaLISA SureFire Ultra Human Total Progesterone Receptor Detection Kit, 10,000 Assay Points
AlphaLISA Surefire Ultra Total Protein
The AlphaLISA™ SureFire® Ultra™ Human Total Progesterone Receptor assay is a sandwich immunoassay for quantitative detection of total Progesterone Receptor in cellular lysates using Alpha Technology.
For research use only. Not for use in diagnostic procedures. All products to be used in accordance with applicable laws and regulations including without limitation, consumption and disposal requirements under European REACH regulations (EC 1907/2006).
| Feature | Specification |
|---|---|
| Application | Protein Quantification |
The AlphaLISA™ SureFire® Ultra™ Human Total Progesterone Receptor assay is a sandwich immunoassay for quantitative detection of total Progesterone Receptor in cellular lysates using Alpha Technology.
For research use only. Not for use in diagnostic procedures. All products to be used in accordance with applicable laws and regulations including without limitation, consumption and disposal requirements under European REACH regulations (EC 1907/2006).
Product Variants
Unit Size: 100 Assay Points
Part #:
ALSU-TPROGR-A-HV
List Price
USD 694.00
Unit Size: 500 Assay Points
Part #:
ALSU-TPROGR-A500
List Price
USD 2,346.00
Unit Size: 10,000 Assay Points
Part #:
ALSU-TPROGR-A10K
List Price
USD 14,270.00
Unit Size: 50,000 Assay Points
Part #:
ALSU-TPROGR-A50K
List Price
USD 46,060.00
AlphaLISA SureFire Ultra Human Total Progesterone Receptor Detection Kit, 10,000 Assay Points
AlphaLISA Surefire Ultra Total Protein
AlphaLISA SureFire Ultra Human Total Progesterone Receptor Detection Kit, 10,000 Assay Points
Product information
Overview
The Progesterone Receptor (PR) is a nuclear hormone receptor activated by the steroid hormone progesterone. PR regulates gene expression involved in reproductive tissue function, cellular differentiation, and proliferation. It exists in two isoforms, PR-A and PR-B, each with distinct and sometimes opposing roles in gene regulation. PR is a critical player in breast cancer development and is a target for endocrine therapies.
The AlphaLISA SureFire Ultra Human Total Progesterone Receptor Detection Kit is a sandwich immunoassay for the quantitative detection of total Progesterone Receptor in cellular lysates, using Alpha Technology.
Formats:
- The HV (high volume) kit contains reagents to run 100 wells in 96-well format, using a 60 μL reaction volume.
- The 500-point kit contains enough reagents to run 500 wells in 384-well format, using a 20 μL reaction volume.
- The 10,000-point kit contains enough reagents to run 10,000 wells in 384-well format, using a 20 μL reaction volume.
- The 50,000-point kit contains enough reagents to run 50,000 wells in 384-well format, using a 20 μL reaction volume.
AlphaLISA SureFire Ultra kits are compatible with:
- Cell and tissue lysates
- Antibody modulators
- Biotherapeutic antibodies
Alpha SureFire Ultra kits can be used for:
- Cellular kinase assays
- Receptor activation studies
- Screening
Specifications
| Application |
Protein Quantification
|
|---|---|
| Automation Compatible |
Yes
|
| Brand |
AlphaLISA SureFire Ultra
|
| Detection Modality |
Alpha
|
| Host Species |
Human
|
| Molecular Modification |
Total
|
| Product Group |
Kit
|
| Shipping Conditions |
Shipped in Blue Ice
|
| Target |
Progesterone Receptor
|
| Target Class |
Phosphoproteins
|
| Target Species |
Human
|
| Technology |
Alpha
|
| Therapeutic Area |
Oncology
|
| Unit Size |
10,000 Assay Points
|
How it works
Total-AlphaLISA SureFire Ultra assay principle
The Total-AlphaLISA SureFire Ultra assay measures the expression level of a protein target in a cell lysate.
The Total-AlphaLISA SureFire Ultra assay uses two antibodies which recognize two different distal epitopes on the targeted protein. AlphaLISA assays require two bead types: Acceptor and Donor beads. Acceptor beads are coated with a proprietary CaptSure™ agent to specifically immobilize the assay specific antibody, labeled with a CaptSure™ tag. Donor beads are coated with streptavidin to capture one of the detection antibodies, which is biotinylated. In the presence of targeted protein, the two antibodies bring the Donor and Acceptor beads in close proximity whereby the singlet oxygen transfers energy to excite the Acceptor bead, allowing the generation of a luminescent Alpha signal. The amount of light emission is directly proportional to the quantity of protein present in the sample.
Total-AlphaLISA SureFire Ultra two-plate assay protocol
The two-plate protocol involves culturing and treating the cells in a 96-well plate before lysis, then transferring lysates into a 384-well OptiPlate™ plate before the addition of Total-AlphaLISA SureFire Ultra detection reagents. This protocol permits the cells' viability and confluence to be monitored. In addition, lysates from a single well can be used to measure multiple targets.
Total-AlphaLISA SureFire Ultra one-plate assay protocol
Detection of Total target protein with AlphaLISA SureFire Ultra reagents can be performed in a single plate used for culturing, treatment, and lysis. No washing steps are required. This HTS designed protocol allows for miniaturization while maintaining AlphaLISA SureFire Ultra quality.
Assay validation
Increase of Progesterone receptor in 17-ß estradiol treated cells
MCF7 cells were seeded in a 96-well plate (40,000 cells/well) in complete medium, and incubated overnight at 37°C, 5% CO2. The cells were treated with 0.78 µM of 17-ß Estradiol for 24 hours.
After treatment, the cells were lysed with 100 µL of Lysis Buffer for 10 minutes at RT with shaking (350 rpm). Progesterone Receptor Phospho (Ser190) and Total levels were evaluated using respective AlphaLISA SureFire Ultra assays. For the detection step, 10 µL of cell lysate (approximately 4,000 cells) were transferred into a 384-well white OptiPlate, followed by 5 µL of Acceptor mix and incubated for 1 hour at RT. Finally, 5 µL of Donor mix was then added to each well and incubated for 1 hour at RT in the dark. The plate was read on an Envision using standard AlphaLISA settings.
As expected, treatment with 17-ß Estradiol caused an increase in both Phospho (Ser190) and Total Progesterone Receptor levels.
Increase of Progesterone receptor in Promegestone treated cells
MCF7 cells were seeded in a 96-well plate (20,000 cells/well) in complete medium, and incubated for 48 hours at 37°C, 5% CO2. The cells were treated with 1 µM of Promegestone for 2 hours. After treatment, the cells were lysed with 100 µL of Lysis Buffer for 10 minutes at RT with shaking (350 rpm). Progesterone Receptor Phospho (Ser190) and Total levels were evaluated using respective AlphaLISA SureFire Ultra assays.
For the detection step, 10 µL of cell lysate (approximately 2,000 cells) were transferred into a 384-well white OptiPlate, followed by 5 µL of Acceptor mix and incubated for 1 hour at RT. Finally, 5 µL of Donor mix was then added to each well and incubated for 1 hour at RT in the dark. The plate was read on an Envision using standard AlphaLISA settings.
As expected, treatment with Promegestone, a well published progesterone receptor agonist, caused an increase in both Phospho (Ser190) and Total Progesterone Receptor levels.
Decrease of Progesterone receptor in fulvestrant treated cells
MCF7 cells were seeded in a 96-well plate (40,000 cells/well) in complete medium, and incubated overnight at 37°C, 5% CO2. The cells were treated with 10 µg/mL Fulvestrant for 18 hours. After treatment, the cells were lysed with 100 µL of Lysis Buffer for 10 minutes at RT with shaking (350 rpm). Progesterone Receptor Phospho (Ser190) and Total levels were evaluated using respective AlphaLISA SureFire Ultra assays.
For the detection step, 10 µL of cell lysate (approximately 4,000 cells) were transferred into a 384-well white OptiPlate, followed by 5 µL of Acceptor mix and incubated for 1 hour at RT. Finally, 5 µL of Donor mix was then added to each well and incubated for 1 hour at RT in the dark. The plate was read on an Envision using standard AlphaLISA settings.
As expected, Fulvestrant triggered a decrease of both Phospho (Ser190) and Total Progesterone Receptor while Total Cofilin levels remained unchanged (data not shown).
Assay versatility
Versatility of total Progesterone receptor assay
Human cell lysates were diluted with Lysis Buffer. Approximate number of cells/datapoint is indicated on graph.
Total Progesterone Receptor levels were evaluated using the AlphaLISA SureFire Ultra assay. For the detection step, 10 µL of cell lysate were transferred into a 384-well white OptiPlate, followed by 5 µL of Acceptor Mix and incubated for 1 hour at RT. Finally, 5 µL of Donor Mix was then added to each well and incubated for 1 hour at RT in the dark. The plate was read on an Envision using standard AlphaLISA settings.
High levels of Total Progesterone Receptor were detected in breast cancer cell lines, MCF7 and BT-474, expression levels were also found in K-562 and BT-20 cell lines.
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