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| 1. |
Hill, K.M., Huang, Y., Yip, S.C., Yu, J., Segall, J.E., and Backer, J.M.
(2001)
N-terminal domains of the class IA phosphoinositide 3-kinase regulatory subunit play a role in cytoskeletal but not mitogenic signaling.
J. Biol. Chem.
276
,
16374-16378
.
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Notes:
The mechanism of lamellipod extension in the metastatic breast cancer cell line MTLn3 cells was studied to determine the signaling pathways involved. The cells were microinjected with a protein-protein interaction domain which inhibits lamellipod extension and incubated in the absence or presence of 1 M sorbitol for 30 minutes. Cells were then fixed in 10% paraformaldehyde, permeabilized with methanol, blocked with 1% bovine serum albumin/5% donkey serum, and immunostained with the Anti-ACTIVE® JNK antibodies to measure JNK activation.
(0002383) |
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Products: Anti-ACTIVE® JNK pAb, Rabbit, (pTPpY) |
| 2. |
Ackerley, S., Grierson, A.J., Brownlees, J., Thornhill, P., Anderton, B.H., Leigh, P.N., Shaw, C.E., and Miller C.C.
(2000)
Glutamate slows axonal transport of neurofilaments in transfected neurons.
J. Cell Biol.
150
,
165-175
.
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Notes:
The authors seek to determine the role of glutamate in excitotoxicity and neurofilament accumulation seen in some neurodegenerative diseases. Neurofilament light, middle, and heavy chains were expressed from rat cDNAs cloned into the pCI-neo Mammalian Expression vector in SW-13 cells. Primary rat cortical neurons were transfected with a neurofilament middle chain and green fluorescent fusion protein. SW13 cells and primary rat cortical neurons were transfected with the ProFection® Mammalian Transfection System–Calcium Phosphate. The CytoTox 96® Non-Radioactive Cytotoxicity Assay was used to monitor glutamate toxicity in these cell. To determine the role of the MAPK and JNK signaling pathways, SW13- cells and primary neuronal cells were immunostained for dually phosphorylated MAPK and JNK using Promega's Anti-ACTIVE® MAPK pAb and Anti-ACTIVE® JNK pAb, respectively. Cells were fixed in 4% paraformaldehyde, permeabilized in 0.1% Triton® X-100 in PBS, blocked with 0.2% Tween® 20 in TBS, and incubated with primary antibodies diluted in blocking solution. Western blot analyses were performed on the primary cortical neurons to quantitate the level of dually phosphorylated MAPK protein The blots were also probed with a pan MAPK antibody that detects total (active and inactive) MAPK.
(0002382) |
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Products: Anti-ACTIVE® JNK pAb, Rabbit, (pTPpY) | Anti-ACTIVE® MAPK pAb, Rabbit, (pTEpY) | CytoTox 96® Non-Radioactive Cytotoxicity Assay | pCI-neo Mammalian Expression Vector | ProFection® Mammalian Transfection System—Calcium Phosphate |
| 3. |
Davis, P.K., Johnson, G.V.W.
(1999)
The microtubule binding of tau and high molecular weight tau in apoptotic PC12 cells is impaired because of altered phosphorylation
J. Biol. Chem.
274
,
35686-35692.
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Notes:
The Anti-ACTIVE® JNK pAb was used for immunocytochemical detection of dually phosphorylated JNK in NGF/serum cultured and deprived PC12 cells. In the normally cultured cells (NGF and serum present) there was light and diffuse staining for the activated JNK throughout the cytoplasm. Upon NGF and serum withdrawal, the staining for the activated JNK is mostly nuclear and coincident with tau staining.
(0001257) |
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Products: Anti-ACTIVE® JNK pAb, Rabbit, (pTPpY) |