| Code | CSB-RA002791A401phHU |
| Size | US$210 |
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| Application | Recommended Dilution |
|---|---|
| WB | 1:500-1:5000 |
| IF | 1:20-1:200 |
BRAF occupies a central position in the RAS-RAF-MEK-ERK signaling cascade, where it functions as a serine/threonine kinase that transduces mitogenic signals from cell surface receptors to downstream effectors controlling proliferation, differentiation, and survival. Phosphorylation at threonine 401 represents a key regulatory modification that influences BRAF kinase activity and its interactions within the MAPK pathway. Given BRAF's prominent role in oncogenic signaling—particularly in melanoma and other malignancies—precise detection of this phosphorylation site provides researchers with critical insight into pathway activation states and therapeutic response mechanisms.
This recombinant rabbit monoclonal antibody (clone 4B9) offers the reproducibility and sequence-defined consistency that phospho-specific detection demands. Because recombinant production eliminates the batch variability inherent in traditional hybridoma methods, researchers can confidently compare phosphorylation dynamics across longitudinal studies or between collaborating laboratories without concerns about antibody drift affecting their results.
Validation studies demonstrate robust performance across multiple experimental platforms. Western blot analysis of A549 lung adenocarcinoma cells reveals specific detection of phospho-BRAF at the expected 85 kDa molecular weight, with comparative analysis of EGF-treated versus untreated lysates enabling assessment of stimulus-dependent phosphorylation changes. The antibody performs effectively at dilutions ranging from 1:500 to 1:5000, providing flexibility to optimize signal-to-background ratios for different sample preparations. Immunofluorescence staining of HepG2 hepatocellular carcinoma cells confirms utility for spatial localization studies, with validated performance at dilutions between 1:20 and 1:200.
This phospho-specific antibody serves researchers investigating MAPK pathway regulation, oncogenic signaling mechanisms, and therapeutic targeting of the RAF kinase family in human cell models.
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