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Lysosomal dysregulation is the hallmark of many diseases including neurodegenerative diseases, lysosomal storages diseases, and aging. The lysosome is responsible for eliminating cellular waste in a multistep pathway called autophagy. This process is well regulated and can be affected by a number of stimulants including nutrient starvation, physiological stress, and chemical induction. Sequestosome-1, also known as p62, is incorporated into autophagosomes then subsequently degraded in the final steps of autophagy. Assessing this protein can help decipher a block or increased flux in autophagy.
Quantification of p62 levels within cellular lysates is often performed with labor intensive wash-based ELISA assays. In this study, chloroquine is used as an inhibitor of autophagy to show how the homogeneous no-wash AlphaLISA® p62 assay can detect changing levels of this lysosomal protein.
Mouse pharmacological models continue to play a large role in the study of human disease, and mouse tool reagents have shown high utility in immunology and cancer research for decades. It can often be quicker to learn about immunology and the regulation of immune responses using a syngeneic mouse model. However, working in mouse systems can often require the development of separate mouse reagents, if the therapeutic agent of interest does not cross-react with mouse. Find out how the AlphaLISA® human PD-1/PD-L1 and AlphaLISA mouse PD-1/PD-L1 binding assays provide a fast, powerful, homogeneous platform for obtain binding potencies from potential novel drug candidates.
Immune checkpoints serve a critical role in the immune system to prevent autoimmunity and manage the degree and duration of an immune response. Cytotoxic T-Lymphocyte-associated protein 4 (CTLA-4 or CD152) is an inhibitory transmembrane protein involved in an immune checkpoint of significant interest for therapeutic development. When CTLA-4 is expressed and competes with CD28, the immune system response is downregulated. As a result of this immune system response balance, immune checkpoints provide an opportunity for therapeutic intervention to modulate immune system activity.
There is a high demand for new drugs to block CTLA-4 and modulate immune system activity. In this application note, we demonstrate how to screen for novel CTLA-4 blocking drugs by utilizing the AlphaLISA CTLA-4/CD80 binding assay.
Tumor Necrosis Factors (TNFs) are cytokines that are the primary modifiers of inflammatory and immune response. Researchers have shown that a soluble form of TNFR1 (sTNFR1) is a truncated version of the receptor produced by the disintegration and extracellular release of membranous protein on the cell surface (ectodomain shedding). sTNFR1 is found in healthy and diseased patients alike, however increased sTNFR1 levels are an indicator for disease states such as inflammation, infection, and asthma.
Here we present a way to measure TNFR1 using homogeneous bead-based AlphaLISA assay. The human TNFR1 AlphaLISA® detection kit was designed for the quantitative determination of soluble TNFR1 in serum and cell culture media. This technical note further demonstrates the functionality of the kit by detecting sTNFR1 in cell supernatant as well as TNFR1 on the cell membrane.