Resources Archive - TGR BioSciences

App Notes

Endogenous detection of toll-like receptor-mediated IRF5 signaling using AlphaLISA SureFire® Ultra. – Application Note

Inflammation is a vital defense mechanism of the immune system, protecting the host against microbial infections and maintaining tissue homeostasis. However, when inflammation is exacerbated, it can drive the progression of various human diseases, including autoimmune and autoinflammatory disorders. At the center of this inflammatory response is interferon regulatory factor 5 (IRF5), a pivotal transcription factor involved in antiviral defense mechanisms and inflammatory pathways that regulate type I interferon (IFN-α and IFN-β) production and pro-inflammatory cytokine responses, including interleukin-6 (IL-6), IL-12, IL-23 and tumor necrosis factor-alpha (TNF-α) [1, 2].

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Poster

Detecting changes in phosphorylation events during the cell cycle with AlphaLISA SureFire Ultra technology. – Technical Note

Lamin A/C is a nuclear protein involved in multiple cellular processes including chromosome organization, transcriptional regulation, DNA repair, cell signalling and cell cycle regulation1. Lamin A/C phosphorylation plays a role in its incorporation into the nuclear lamina as well as disassembly during mitosis2. Phosphorylation at the so-called “mitotic sites” (Ser22 and Ser392) reaches the maximum level at the onset of mitosis and is triggered by the complex formed by CDK1 and Cyclin B1. The phosphorylation/dephosphorylation events surrounding the CDK1-Cyclin B1-Lamin A/C pathway are associated with alterations in the cell cycle that have an impact not only on the cell morphology and structure but also on the cell fate3. The dysregulation in the levels of these phosphorylated proteins and their total counterparts has been associated with increased cell growth, genomic instability and cell senescence in different types of cancers, becoming a key area for the discovery of efficient therapies to prevent tumor progression1,2

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App Notes

PROTAC degraders targeting cyclin kinases with AlphaLISA Surefire Ultra screening technology for study of cell cycle regulation in oncology. – Technical Note

The cell cycle is intricately regulated by a network of proteins, with Cyclin D1 playing a pivotal role in initiating cell cycle progression. Cyclin D1 forms complexes with Cyclin-Dependent Kinase 4 (CDK4) and CDK6 during the G1 phase, promoting the phosphorylation of the retinoblastoma protein (Rb).1 This phosphorylation releases the inhibitory influence of Rb on the E2F transcription factors, allowing the expression of genes necessary for cell cycle entry. Subsequently, Cyclin E1 associates with CDK2, forming the Cyclin E/CDK2 complex, further contributing to Rb phosphorylation and progression through the G1 phase into the S phase.2......

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App Notes

Protein phosphorylation and cellular signaling cascades within integrated stress response pathway with AlphaLISA SureFire Ultra assays. – Technical Note

Protein phosphorylation is a critical regulatory mechanism for many cellular processes and is tightly controlled by kinase and phosphatase cascades. Therefore, monitoring phosphorylation events in a cellular model can be a useful approach to study the activity of compounds, as well as understand their mechanism of action or target modulation. Here we demonstrate the utility of AlphaLISA™ SureFire™ Ultra™ (ALSU) assays to reliably measure a targeted phosphorylation event and total protein levels in cell-based experiments with a focus on the eIF2α/ATF-4 pathway. ...

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Poster

JAK/STAT signalling in PBMCs: High sensitivity AlphaLISA SureFire® Ultra assays for measuring JAK phosphorylation and downstream activation/inhibition of STAT family proteins SLAS-2025

AlphaLISA™ SureFire® Ultra™ assays are traditionally known for measuring target phosphorylation and downstream cascade activation in endogenous cell systems. Here we showcase new SureFire assay data for 3 JAK family members: Phospho/Total JAK1, JAK2 and TYK2. High signal windows were generated using heathy human peripheral blood mononuclear cells (PBMCs) showing IFNα, IFNγ and IL-4-mediated activation of JAK proteins combined with the downstream phosphorylation of STAT1, STAT3, STAT5 and STAT6 family members. In addition, specific JAK inhibitor (Ruxolitinib & Pacritinib) data was generated in the immortalised HEL 92.1.7 and THP-1 cell lines using this broad suite of SureFire JAK/STAT assays.

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Poster

Novel and highly sensitive readouts for Toll-like Receptor (TLR) activation in endogenous cells: Specific detection of IRF5 Phospho (S446) and Dimer with AlphaLISA™ SureFire® Ultra™ technology SLAS-2025

Interferon regulatory factor 5 (IRF5) is a key transcription factor for the activation of the pathogen-induced innate and acquired immune responses. IRF5 exists in an inactive monomeric form in the cytosol. Following TLR activation IRF5 undergoes conformational changes prior to phosphorylation and dimerization after which it is translocated to the nucleus regulating the transcription of pro-inflammatory cytokines. Deregulation of IRF5 can lead to the unrestricted production of interferons leading to numerous inflammatory and autoimmune diseases. As a result, IRF5 has emerged as a key therapeutic target for arresting the pathogenesis associated with an unregulated IRF5 pathway.....

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