Human Apoptosis 3-Plex kit
MILLIPLEX
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.
Multiplexing
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.
Human
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.
The T-cell Receptor (TCR) is found on the surface of T-cells and is responsible for recognition of foreign antigen. Antigen binding to the TCR results in T-cell activation and proliferation (clonal selection). The TCR is composed of an α and a β subunit, either CD4 (on Th cells) or CD8 (on Tc cells), and CD3, which is made up of six subunits arranged into three dimers derived from five proteins (δ, ε, γ, ζ and η). The TCR initiates a signaling cascade that begins with the CD3 complex and proceeds through Lck to LAT and so on to Ras and then ERK, or through Fyn and ZAP-70 and so on to Rac and then JNK.
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.
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
Kit capacity: 100 Assay Points
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3-Plex Apoptosis, beads (Cat #42-670)
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3-Plex Apoptosis, biotin (Cat #44-670)
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Lysis Buffer (Cat.# 43-040)
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Assay Buffer 1 (Cat.# 43-010)
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Jurkat Cell lysate: unstimulated (Cat #47-206)
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Jurkat Cell lysate: Anisomycin (Cat #47-207)
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Streptavidin-Phycoerythrin (Cat.# 45-001D)
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96-well filter plate
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Empty mixing vials
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Cancer Kits
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Cell Signaling Kits
Multiplexing
Luminex xMAP
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Cancer Kits
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Cell Signaling Kits
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Multiplex Assays
Multiplex Assays
