Macrophage Chemokine Ligand 3 (CCL3) ELISA Kit, qPCR (MTS-1123-HM27)

It depends on your research question. qPCR measures CCL3 mRNA and is excellent for detecting early transcriptional responses and pathway activation. However, mRNA levels do not always translate directly into secreted protein due to regulation at translation and secretion stages. If your conclusions involve functional chemokine availability or receptor signaling, a protein ELISA is recommended as confirmation. Many researchers use qPCR to screen conditions quickly, then validate key findings at the protein level. Combining both can clarify whether changes are transcriptional only or reflected in secreted CCL3.

Use validated housekeeping genes that are stable under your polarization conditions; macrophage activation can change common reference genes. We recommend evaluating at least two candidate reference genes and selecting those with minimal variation across treatments. Normalize consistently using ΔCt/ΔΔCt methods and include technical replicates. Keep RNA input and reverse transcription conditions uniform. Also include no-template and no-RT controls to detect contamination. For publication-ready work, report primer/probe performance indicators such as efficiency and specificity checks to strengthen confidence.

Macrophage samples and tissue-derived materials can carry inhibitors (lipids, heme, polysaccharides). Use robust RNA extraction with adequate wash steps, and if needed, add an extra purification step. Assess RNA integrity and purity (e.g., absorbance ratios) and consider modest dilution if inhibition is suspected. Set up reactions with master mixes, use calibrated pipettes, and avoid repeated freeze-thaw of RNA. If Ct variability persists, check for genomic DNA contamination and confirm amplification specificity via melt curve or probe-based design.