Journal: The Journal of Neuroscience

Article Title: Fas/Apo [Apoptosis]-1 and Associated Proteins in the Differentiating Cerebral Cortex: Induction of Caspase-Dependent Cell Death and Activation of NF-κB

doi: 10.1523/jneurosci.19-05-01754.1999

Figure Lengend Snippet: Figure 6. The expression of the Fas-associated proteins RIP (A), FADD (B), FLIP (C), and FasL (D) was examined by Western analysis of 50 mg of detergent-soluble protein obtained from P6 cerebral cortex obtained from four separate animals (lanes 1-4). Bands corresponding to RIP (74 kDa) and FasL (37 kDa) were detected in 10 mg of Jurkat cell lysates (a positive control) and in P6 cortex. FLIP expression in P6 cortex was confirmed by the appearance of a similar band just below the 66 kDa molecular weight marker, corresponding to the expected size of ;60 kDa, in protein extract from COS7 cell transfected with the full-length FLIP plasmid (positive control). In contrast, we did not detect any bands corresponding to the expected size for FADD in the cerebral cortex, although a band of the approximate size (24 kDa) was observed in Jurkat cell-derived protein (a positive control). Mkr, Biotinylated marker.

Article Snippet: Binding of the second, primary antibody was visualized by the appropriate secondary antibody, either a biotinylated donkey anti-rabbit antibody (Jackson ImmunoResearch, 1:500) or horse anti-mouse (Vector Laboratories, 1:1000) coupled to either streptavidin-FITC or rhodamineavidin.

Techniques: Expressing, Western Blot, Positive Control, Molecular Weight, Marker, Transfection, Plasmid Preparation, Derivative Assay