Revision 3
#92657
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For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:
W, W-S, IP, IF-F, IF-IC
Reactivity:
H M R
Sensitivity:
Endogenous
MW (kDa):
49
Source/Isotype:
Rabbit IgG
UniProt ID:
#Q13562
Entrez-Gene Id:
4760
Product Usage Information
| Application | Dilution |
|---|---|
| Western Blotting | 1:1000 |
| Simple Western™ | 1:10 - 1:50 |
| Immunoprecipitation | 1:100 |
| Immunofluorescence (Frozen) | 1:50 |
| Immunofluorescence (Immunocytochemistry) | 1:100 |
Storage
Specificity/Sensitivity
Source / Purification
Background
Erk1/2 kinase activity is stimulated by glucose and calcium influx in pancreatic beta cells, and the subsequent phosphorylation of NeuroD1 at Ser274 promotes insulin gene transcription (7,8). Phosphorylated NeuroD1 in the pancreas is increasingly translocated to the nucleus, where it directly interacts with the insulin promoter (9). Active, nuclear NeuroD1 acts in synergy with Pdx1 to activate insulin, NKX2.2, and islet amyloid polypeptide (IAPP) transcription while suppressing Pdx1-driven transcription of somatostatin (10,11). In developing rat pineal gland, phosphorylation of NeuroD1 at Ser274 and Ser336 is tightly correlated with NeuroD1 nuclear localization, and cytoplasmic to nuclear shuttling of the protein is under cyclical day and night adrenergic control (12). In 293T cells, transfection of a nonphosphorylatable Ser274Ala mutant blocked NeuroD1 ubiquitination and proteasomal degradation (13). The localization of phosphorylated NeuroD1 is reversed in these cells compared to pancreatic beta cells, with a Ser274Glu phospho-mimic localized primarily to the cytoplasm in the former, further evidence of the context-specific nature of this modification. (13).
Background References
- Schonhoff, S.E. et al. (2004) Endocrinology 145, 2639-2644.
- Sharma, A. et al. (1999) Mol. Cell Biol. 19, 704-713.
- Chae, J.H. et al. (2004) Mol. Cells 18, 271-288.
- Gaudillière, B. et al. (2004) Neuron 41, 229-241.
- Miyata, T. et al. (1999) Genes Dev. 13, 1647-1652.
- Naya, F.J. et al. (1997) Genes Dev. 11, 2323-2334.
- Khoo, S. and Cobb, M.H. (1997) Proc Natl Acad Sci USA 94, 5599-604.
- Khoo, S. et al. (2003) J Biol Chem 278, 32969-77.
- Petersen, H.V. et al. (2002) FEBS Lett 528, 241-5.
- Itkin-Ansari, P. et al. (2005) Dev Dyn 233, 946-53.
- Babu, D.A. et al. (2008) J Biol Chem 283, 8164-72.
- Castro, A.E. et al. (2015) J Pineal Res 58, 439-51.
- Lee, T.Y. et al. (2020) Exp Neurobiol 29, 189-206.
Species Reactivity
Species reactivity is determined by testing in at least one approved application (e.g., western blot).
Western Blot Buffer
IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
Applications Key
W: Western Blotting W-S: Simple Western™ IP: Immunoprecipitation IF-F: Immunofluorescence (Frozen) IF-IC: Immunofluorescence (Immunocytochemistry)
Cross-Reactivity Key
H: Human M: Mouse R: Rat
Trademarks and Patents
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.
Limited Uses
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Revision 3
Revision 3
Revision 3
