Human Phospho-SMAD4 (Thr277) and Total SMAD4 PharmaGenie ELISA Kit (SBRS1973)

Built around a dual-analyte configuration, the Phospho-SMAD4 (Thr277) and Total SMAD4 PharmaGenie ELISA Kit from Assay Genie quantifies phosphorylated SMAD4 at threonine 277 and total SMAD4 in the same wells, so researchers can normalize phosphorylation signal directly to total protein content. The Thr277 site lies in the SMAD4 linker region and is modulated through TGF-β, BMP, and MAPK pathway activity, supporting studies of receptor serine/threonine kinase signaling, tumor-suppressor function, and downstream transcriptional responses in pancreatic, colorectal, and other gastrointestinal cancer models where SMAD4 loss-of-function is clinically relevant. The kit is applied to cell lysates or tissue lysates prepared from cultured lines or xenograft/biopsy material to profile pathway activation under cytokine stimulation, small-molecule inhibitor treatment, or genetic perturbation (CRISPR, siRNA, overexpression). A 100 µL sample volume is loaded into pre-coated microplate wells, and the included phospho-specific and pan-SMAD4 capture antibodies resolve the two readouts in parallel. For run-to-run reliability, reconstituted reagents should be aliquoted to limit freeze-thaw cycles and the working conjugate should be protected from light until use. This dual-detection format is widely adopted for epithelial-mesenchymal transition studies, BMP response profiling, and juvenile polyposis syndrome mechanism research. Intended for research use only.