CD154 iFluor 488

Overview

CD154 iFluor 488 is a Mouse monoclonal targeting CD154, supplied as a iFluor™ 488 format for FC workflows. It supports measurement of Human target expression in common experimental systems.

Key elements and design rationale

• Clone: 5C8 — consistent clone identity can support panel reproducibility and cross-study comparisons.
• Isotype: IgG2a, k — informs selection of matched controls and secondary reagents when relevant.
• Conjugate: iFluor™ 488 — enables direct detection in fluorescence-based assays. Excitation is typically matched to Blue (488nm) lasers in cytometer configurations.
• Host species: Mouse — useful for panel design and control strategy planning.
• Reactivity: Human — interpret staining in the context of species-specific sequence and expression differences.

Key specifications such as clone identity, isotype, and fluorophore conjugation help researchers align panel design, control selection, and instrument configuration with the biological question and sample type.

Biological background

The clone 5C8, a mouse monoclonal antibody, specifically recognizes a ~30-33 kDa activation-induced human CD4+ T cells surface molecule known as CD154 or CD40 ligand (CD40L). The CD40 interacts with CD154 through T cell-B cell activating molecule, gp39, TNF cytokine related activation protein. CD154 is primarily expressed on the surface of activated CD4+ T cells but can also be expressed by platelets, mast cells, macrophages, basophils, NK cells, B cells, CD8+ T cells as well as non-hematopoietic cells including smooth muscle cells, endothelial cells, and epithelial cells. CD154 plays important roles in the pathogenesis of certain blood cancer such as the CD4+ T cells of samples with CLL fail to express surface CD154 antigen even after CD3 ligation, and proximal and distal activation events are required for the neonatal CD4+ T cells for the CD154 gene transcription to establish the normal immune system.

Research relevance and current trends

• High-parameter immunophenotyping: combining CD154 with complementary lineage and activation markers to resolve complex cell states.
• Panel standardization and data comparability: increasing emphasis on consistent reagents, compensation-aware fluorophore choices, and shared gating strategies.
• Integration with single-cell multi-omics: pairing surface marker profiling with transcriptomic or proteomic readouts to connect phenotype to function.

Common research applications

• Flow cytometry: quantify CD154-positive populations and compare expression distributions across conditions or time points.
• Cell sorting: enrich CD154-defined subsets for downstream RNA/protein assays or functional readouts.

Changes in measured signal are typically interpreted in the context of cell subset frequency, activation/differentiation state, and sample processing effects rather than as a standalone readout.

Notes for experimental interpretation

• Fluorophore selection: consider brightness, spectral overlap, and instrument configuration; compensation and spillover can affect apparent population boundaries.
• Biology-driven confounders: activation state, differentiation, and isoform/PTM variation can shift epitope accessibility and apparent expression.
• Control concepts: include matched isotype and fluorescence-minus-one (FMO) controls where appropriate, and interpret results alongside biological positive/negative reference samples.

For antibody-based assays, monoclonal versus polyclonal format can influence epitope recognition breadth and signal consistency. Conjugated antibodies support direct detection and can simplify multicolor panel design when paired with appropriate controls and instrument settings.

Customization & Add-ons: Can’t find the antibody you need—or require a custom format for your assay? We can help you source the best match or support custom antibody solutions for diverse research needs, including species and isotype selection, conjugations and labeling (e.g., HRP/AP, biotin, fluorophores), purification grade options (Protein A/G, affinity purified), formulation preferences (buffer selection, carrier-free, glycerol-free), custom concentrations and aliquoting, low-endotoxin options for cell-based work, and application-focused QC/validation support (project dependent). Click Talk to a Scientist to submit a request, email us at support@biohippo.com, or explore our Research Services for additional support—our team will follow up with feasibility details and next steps.