MILLIPLEX MAP Human Apoptosis 3-Plex kit
Description:
MILLIPLEX MAP Human Apoptosis 3-Plex kit
Trade Name:
MILLIPLEX
Product Overview:
3-plex Apoptosis Signaling kit is used to detect changes in activated Caspase 3 and PARP in cell lysates using the Luminex® system. Total GAPDH is provided in the kit for total protein normalization. The detection assay is a rapid, convenient alternative to Western Blotting and immunoprecipitation procedures. Each kit has sufficient reagents for 100 individual assays.
Key Applications:
Multiplexing
Application Notes:
Specificity - Immunoprecipitation/Western Blot Analysis of Multiplexed Analytes in Jurkat Cells
20 mg of lysates were mixed with capture antibody beads to immunoprecipitate each respective protein. The immunoprecipitated proteins were separated on SDS-PAGE, transferred to nitrocellulose, and probed with biotin labeled active (Cleaved PARP and Caspase 3) or total (GAPDH), reporter antibodies. The proteins were imaged using Streptavidin-HRP and chemiluminescence.
20 mg of lysates were mixed with capture antibody beads to immunoprecipitate each respective protein. The immunoprecipitated proteins were separated on SDS-PAGE, transferred to nitrocellulose, and probed with biotin labeled active (Cleaved PARP and Caspase 3) or total (GAPDH), reporter antibodies. The proteins were imaged using Streptavidin-HRP and chemiluminescence.
Species Reactivity:
Human
Usage Statement:
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Kit or Assay Type:
- Cancer Kits
- Cell Signaling Kits
- Multiplex Assays
Therapeutic Areas:
- Oncology
- Auto-Immuno
Configuration:
Premixed
Components:
- 3-Plex Apoptosis, beads (Cat #42-670)
- 3-Plex Apoptosis, biotin (Cat #44-670)
- Lysis Buffer (Cat.# 43-040)
- Assay Buffer 1 (Cat.# 43-010)
- Jurkat Cell lysate: unstimulated (Cat #47-206)
- Jurkat Cell lysate: Anisomycin (Cat #47-207)
- Streptavidin-Phycoerythrin (Cat.# 45-001D)
- 96-well filter plate
- Empty mixing vials
Premixed or Single Plex:
Yes
Sensitivity:
Multiplex Analysis of Jurkat Cells Treated with Anisomycin - Jurkat Cells were stimulated with 25 µM of anisomycin for 4 hours. The cells were lysed in Millipore Cell Signaling Universal Lysis Buffer containing protease inhibitors. 1 mg of lysate diluted in Millipore Cell Signaling Assay Buffer 1 was analyzed according to the Assay protocol (lysate incubation at 4°C). The Median Fluorescence Intensity(MFI) was measured with the Luminex® system. Each graph below demonstrates that the analytes is both inducible and titratable, indicating the assay is specific.
Panel Type:
Cell Signaling
Detection Methods:
- Fluorescent
- Luminex xMAP
Packaging:
Kit capacity: 100 Assay Points
Materials Required but Not Delivered:
Microbead Sonicator
MultiScreenHTS Vacuum Manifold (Catalogue No. MSVMHTS00)
Luminex® 200 IS System (Catalogue No. 40-009)
Cell lysates or cell extracts harboring proteins of interest
Vortex Mixer
Plate Shaker
Timer
Variable Volume (5-200µL) pipette + tips
Protease inhibitors
MultiScreenHTS Vacuum Manifold (Catalogue No. MSVMHTS00)
Luminex® 200 IS System (Catalogue No. 40-009)
Cell lysates or cell extracts harboring proteins of interest
Vortex Mixer
Plate Shaker
Timer
Variable Volume (5-200µL) pipette + tips
Protease inhibitors
Analytes Available:
Caspase 3 (Active), GAPDH (Total), PARP (Cleaved)