Notes: NF-E2-related factor-2 (NRF2) upregulates transcription of antioxidant response element (ARE)-driven genes. The authors examined the sensitivity of Nrf-2^–/^– primary mouse neurons to oxidative stress-induced apoptosis using the mitochondrial toxins 1-methyl-4-phenyl-1,2,5,6-tetrahydropyridine (MPP⁺) and rotenone. Primary cultures were >090% neurons, as judged by immunostaining with the Anti-βIII Tubulin mAb and other neuron-specific antibodies. Neuronal cytotoxicity in the presence of MPP⁺ and rotenone was monitored using the CellTiter 96^® AQueous Non-Radioactive Cell Proliferation Assay and TUNEL assays. The level of expression of the antioxidant-responsive gene quinone oxidoreductase was measured by mRNA quantitation using RT-PCR, enzyme activity assays and immunocytochemistry. The Reverse Transcription System was used to synthesize cDNA for subsequent amplification by PCR. (3570)

Notes: Rat skin sections were subjected to immunohistochemistry with the Anti-βIII Tubulin mAb to detect metabolic glutamate receptor expressing neurons. Twenty micron sections were fixed in acetone, permeabilized with 0.1% Triton® X-100, and incubated with the Anti-βIII Tubulin mAb at a final concentration of 1µg/ml. (2387)

Notes: During apoptosis, Poly (ADP-ribose) polymerase (PARP) is cleaved by caspase-3 to generate an 85 kDa fragment (p85). An affinity-purified polyclonal antibody to the p85 fragment of PARP is characterized. In Western blot analysis with Jurkat cells treated with an anti-Fas antibody and with SH-Sy5Y cells treated with staurosporine, the antibody recognizes the 85-kDa (p85) fragment of PARP but not full-length PARP. The antibody was used at a concentration of 0.75 µg/ml for Western blots and at 2.5µg/ml for immunocytochemistry. TUNEL labeling of apoptotic Jurkat cells was performed with the DeadEnd™ Fluorometric TUNEL System. Immunocytochemistry was also performed using the Anti-β III Tubulin mAb. Good experimental details are given. (2355)

Notes: Hyperphosphorylation of protein tau in the brain has been linked to Alzheimer's disease. The level of phosphorylated and unphosphorylated protein tau in brains and spinal cords of transgenic mice expressing a protein tau kinase, was quantified by Western blots. Western blot results for protein tau were normalized for tubulin using the Anti-βIII Tubulin mAb.  (2357)

Notes: A primary cell line from rat subventricular zone which remains undifferentiated over time was developed as a model for MAPK activation. Cell were stimulated with 20 ng/ml bFGF and 20 ng/ml EGF (Promega) to activate MAPK. The MAPK pathway was inhibited with either U0126 (Promega) or  PD98059. Immunocytochemistry was performed with the Anti-ERK 1/2 pAb (1:100 dilution) to detect both active and inactive forms of MAPK proteins and with the Anti-ACTIVE™ MAPK pAb (1:100 dilution) to specifically detect the dually phosphorylated, active forms of MAPK. Cells were also immunostained with the neuron specific marker Anti-III-tubulin mAb (0.5 µg/ml). Cell proliferation was monitored with the CellTiter 96® AQueous One Solution Cell Proliferation Assay System. Apoptosis within the cell population was monitored using the DeadEnd™ Fluorometric TUNEL System. (2391)

Notes: The role of the ataxin-1 protein in spinocerebellar ataxia-1 was examined by monitoring the expression levels of various proteins associated with the disorder. βIII-tubulin was used as a neuronal markers in immunofluorescence studies of formalin fixed paraffin embedded sections of mouse brain. The Anti-βIII Tubulin mAb was used at a 1:2000 dilution. (2386)

Notes: Expression of GDNF via an adenovirus vector helped to prevent hippocampal neuron death following transient ischemia. Neurons were detected by immunohistochemistry of gerbil brain sections using an antibody raised against the neuron specific marker βIII tubulin. Animals were perfused with 4% paraformaldehyde and brains were paraffin embedded. Three micron tissue sections were incubated with a 1:300 dilution of Promega's Anti-βIII Tubulin mAb. (2388)