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| 1. |
Ochi, H., Ogino, H., Kageyama, Y. and Yasuda, K.
(2003)
The stability of the lens-specific Maf protein is regulated by Fibroblast Growth Factor (FGF)/ERK signaling in lens fiber differentiation.
J. Biol. Chem.
278
,
537-544
.
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Notes:
Fibroblast growth factor (FGF) signaling is necessary for proliferation and differentiation of chicken lens cells. The transcription factor L-Maf is a lens differentiation factor that appears to mediate FGF signaling. This paper shows that L-Maf is repressed by FGF/ERK signaling and that L-Maf is phosphorylated by ERK. Both Anti-ACTIVE MAPK® pAb and Anti-ERK1/2 pAb were used to determine MAPK levels by Western blotting. Anti-ACTIVE® MAPK pAb was also used in immunocytochemistry analysis of cultured chick lens cells.
(0002773) |
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Products: Anti-ACTIVE® MAPK pAb, Rabbit, (pTEpY) | Anti-ERK 1/2 pAb, Rabbit |
| 2. |
Cottom J., Salvador, L.M., Maizels, E.T., Reierstad, S., Park, Y., Carr, D.W., Davare, M.A., Hell, J.W., Palmer, S.S., Dent, P., Kawakatsu, H., Ogata, M. and Hunzicker-Dunn, M.
(2003)
Follicle-stimulating hormone activates extracellular signal-regulated kinase but not extracellular signal-regulated kinase kinase through a 100-kDa phosphotyrosine phosphatase.
J. Biol. Chem.
278
,
7167-7179
.
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Notes:
This paper investigates the pathway by which follicle stimulating hormone (FSH) activates ERKs (MAPK) p42 and p44 in rat primary granulosa cells. The phosphorylation state of MAPK p42/p44 was assessed by Western blotting using the Anti-ACTIVE® MAPK pAb. Phosphorylated MAPKs were localized to the nucleus of granulosa cells by immunocytochemistry using Anti-ACTIVE® MAPK pAb. Cells were stimulated, fixed with 3.7% formaldehyde, permeabilized with 1% Triton X-100 in PBS, washed, blocked for 1 hour in 1% bovine serum albumin in PBS and incubated overnight at 4°C with a 1:100 dilution of the antibody in PBS containing 1% bovine serum albumin.
(0002768) |
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Products: Anti-ACTIVE® MAPK pAb, Rabbit, (pTEpY) |
| 3. |
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 |
| 4. |
Wahab, N.A., Parker, S., Sraer, J.D., and Mason, R.M.
(2000)
The decorin high glucose response element and mechanism of its activation in human mesangial cells.
J. Am. Soc. Nephrol.
11
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1607-19
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Notes:
Hyperglycemia seems to be involved in the pathogenesis of diabetic nephropathy. The authors identify regulatory promoter elements involved in gene regulation under hyperglycemic conditions and characterize some of the signaling pathways involved. Immunocytochemistry of human mesangial cells grown under normal and high glucose conditions was performed with the Anti-ACTIVE® MAPK pAb to localize dually phosphorylated, active MAPK protein. Cells were fixed in 3.7% paraformaldehyde, permeabilized with 0.1% Triton® X-100 and stained with the Anti-ACTIVE® MAPK pAb overnight at 4°C, or for 1 hour at 37°C. Gel shift assays were performed to identify a regulatory element which appears to be responsive to glucose concentration. Gel shift assays were carried out using Promega's Gel Shift Assay System.
(0002394) |
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Products: Anti-ACTIVE® MAPK pAb, Rabbit, (pTEpY) | Gel Shift Assay Core System | Gel Shift Assay System |
| 5. |
Learish, R.D., Bruss, M.D., and Haak-Frendscho, M.
(2000)
Inhibition of mitogen-activated protein kinase kinase blocks proliferation of neural progenitor cells.
Brain Res. Dev. Brain Res.
122
,
97-109
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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.
(0002391) |
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Products: Anti-βIII Tubulin mAb | Anti-ACTIVE® MAPK pAb, Rabbit, (pTEpY) | Anti-ERK 1/2 pAb, Rabbit | CellTiter 96® AQueous One Solution Cell Proliferation Assay | DeadEnd™ Fluorometric TUNEL System | Donkey Anti-Rabbit IgG (H+L), HRP | MEK Inhibitor U0126 | RQ1 RNase-Free DNase |