IFNβ (mouse) AlphaLISA Detection Kit, 100 Assay Points

Orthogonal systems to cell-based assays are a key requirement in EMA/FDA guidelines for potency estimations and require cross-validation with complementary approaches to prove and strengthen the reliability of results.

In this application note published in collaboration with IBR Inc., you will learn:

• Why Alpha technology represents an ideal cell-free orthogonal system for potency assays
• How AlphaLISA^® assays can be used to determine Bevacizumab/VEGF165 potency
• An example of how to run an AlphaLISA potency assay and the type of data that can be generated
• Why it is suitable for assessing lot-to-lot consistency and equivalence of Bevacizumab and biosimilars

Alpha (Amplified Luminescent Proximity Homogeneous Assay) technology is a bead-based, no-wash alternative to traditional ELISAs. Instead of detection with an HRP substrate, the Alpha assay signal is generated by the excitation of an Eu+-coated bead that has been conjugated to the detection antibody.

Alpha technology offers a simple, straight forward workflow. No wash needed!

1. Add sample
2. Add Acceptor bead mix; incubate 1 hr
3. Add Donor beads; incubate 30 mins
4. Read on Alpha-enable microplate reader

Download this application note to see how no-wash AlphaLISA^® technology provides a more-convenient alternative to ELISA for quantitation of biomarkers in complex sample types, including tissue, serum, and plasma.

Breast cancer tumors can adapt to immune cell infiltration by responding to the increased concentration of interferon gamma (IFN-ɣ) and other cytokines secreted by subsets of T lymphocytes with the upregulation of the immune checkpoint proteins such as Programmed cell death ligand 1 (PD-L1). These checkpoint proteins allow the tumors to evade immune targeting and reduce the immune response, thus promoting tumor progression.

In this application note, you will learn:

• How exogenous addition of IFN-ɣ effects the expression of PD-L1 and secretion of several cytokines in cultures of HCC38 cells (a triple-negative breast cancer cell line)
• How co-culturing activated immune cells and breast cancer cells stimulates differential expression of some immune checkpoint and inflammatory biomarkers compared to culturing cells alone with PBMC-conditioned media
• How to rapidly measure multiple biomarkers in cell culture supernatant and lysates from the same wells of a culture dish to examine protein expression profiles from cancer and immune cell culture models using AlphaLISA^® assays together with ATPlite^™ 1step and the EnVision^® multimode plate reader.

Various cytokines are secreted during an active immune response that can have modulatory effects on target cell populations, including interferon gamma (IFN-ɣ), tumor necrosis factor alpha (TNFa) and several interleukins.

In this application note, you will learn how we investigated:

• The effects of secreted factors on a triple-negative breast cancer cell line by treatment with conditioned media from activated PBMCs
• Whether the modulatory effects of cytokines secreted by infiltrating immune is different when studied in 3D cell cultures
• How to detect and quantify multiple immune-modulated proteins from the same wells in complex cell models grown in 3D spheroid microplates and traditional 2D cell culture using AlphaLISA^® technology and EnVision^® multimode plate reader in a workflow with ATPlite^™ 1step and Opera^® Phenix high-content screening system

Product brochure for the Alpha Technology, a versatile, no wash, homogeneous assay technology that's suitable for a broad range of applications.

Find out about our range of integrated solutions for drug discovery screening in this e-brochure.

Our screening solutions for high-throughput screening, phenotypic screening and data analysis help to streamline drug discovery workflows in labs across the globe. Our portfolio includes automated liquid handling, assays and reagents, imaging and detection systems, and informatics.

Working independently or together, with our solutions you can achieve consistent and accurate results. By accelerating the identification and characterization of effective and safe drug candidates, the PerkinElmer portfolio enables you to optimize efficiency in your lab and deliver more actionable, real-world results.

Download the brochure to learn more about how we can partner with you so that you can discover smarter, more effective, data-driven breakthroughs in the critical screening stages of drug discovery and development.

This guide presents the simple conversion of an ELISA or other immunoassay to an AlphaLISA® immunoassay.