Revvity, ALSU-PLAM-A50K, AlphaLISA Human Phospho-Lamin A/C (Ser22) Detection Kit, 50,000 Assay Points

Overview
Lamins are structural components of the nuclear lamina, a protein network underlying the inner nuclear membrane that determines nuclear shape and size. They can be classified as A type and B type. The B-type lamins are expressed in most cell types, whereas A-type lamins, which include lamin C, are generally expressed in differentiated tissues. Lamins are multi-functional proteins, which play important roles in nuclear structure, chromatin organization, gene positioning, DNA replication and repair, as well as cell proliferation, differentiation, and stress responses. Muscular dystrophies, neuropathies, cardiomyopathies, and syndromes of premature aging can all be caused by mutations in the LMNA gene, which codes for lamins A and C. These disorders are collectively referred to as laminopathies.
The AlphaLISA SureFire Human Phospho-Lamin A/C (Ser22) Detection Kit is a sandwich immunoassay for the quantitative detection of phospho-Lamin A/C (Ser22) 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 kits can be used for:

Cellular kinase assays
Receptor activation studies
Screening
How it works
Phospho-AlphaLISA SureFire Ultra assay principle

The Phospho-AlphaLISA SureFire Ultra assay measures a protein target when phosphorylated at a specific residue.

The assay uses two antibodies which recognize the phospho epitope and a distal epitope 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 phosphorylated 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 phosphoprotein present in the sample.

 

Phospho-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 Phospho-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.

Phospho-AlphaLISA SureFire Ultra one-plate assay protocol

Detection of Phosphorylated 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.
Application-Cell Signaling
Automation Compatible-Yes
Brand-AlphaLISA SureFire Ultra
Detection Modality-Alpha
Lysis Buffer Compatibility-Lysis Buffer B
Molecular Modification-Phosphorylation
Product Group-Kit
Sample Volume-10 µL
Shipping Conditions-Shipped in Blue Ice
Target-Lamin A/C
Target Class-Phosphoproteins
Target Species-Human
Technology-Alpha
Therapeutic Area-Cardiovascular Neuroscience
Unit Size-50,000 assay points