Cell Signaling Technology

Product Pathways - Neuroscience

NeuroD (D35G2) Rabbit mAb #4373

Applications Reactivity Sensitivity MW (kDa) Isotype
W IP ChIP H M R Endogenous 49 Rabbit IgG

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  ChIP=Chromatin IP
Reactivity Key:  H=Human  M=Mouse  R=Rat
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

NeuroD (D35G2) Rabbit mAb detects endogenous levels of total NeuroD protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide surrounding Gln15 of human NeuroD protein.

Western Blotting

Western Blotting

Western blot analysis of extracts from IMR-32 cells using NeuroD (D35G2) Rabbit mAb.

Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 SH-SY5Y cells treated with Human Brain-Derived Neurotrophic Factor (BDNF) #3897 (50 ng/ml) for 30 minutes and either 10 μl of NeuroD (D35G2) Rabbit mAb or 2 μl of Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human EGR1 Promoter Primers #5549, human EGR2 promoter primers, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.

Background

NeuroD is a member of the basic helix-loop-helix (bHLH) family of transcription factors. These proteins function by forming heterodimers with E-proteins and binding to the canonical E-box sequence CANNTG (1,2). Neuronal activity results in CaMKII-mediated phosphorylation of NeuroD at Ser336, which is necessary for formation and growth of dendrites (3,4). NeuroD is also phosphorylated at Ser274 though the results are context dependent as phosphorylation by Erk stimulates NeuroD activity in pancreatic β-cells while phosphorylation by GSK-3β inhibits NeuroD in neurons (3). NeuroD is crucially important in both the pancreas and developing nervous system, and plays a large role in the development of the inner ear and mammalian retina (3). Mice lacking NeuroD become severely diabetic and die shortly after birth due to defects in β-cell differentiation (2,3,5,6). The lack of NeuroD in the brain results in severe defects in development (5). Human mutations have been linked to a number of types of diabetes including type I diabetes mellitus and maturity-onset diabetes of the young (1,3).

  1. Schonhoff, S.E. et al. (2004) Endocrinology 145, 2639-2644.
  2. Sharma, A. et al. (1999) Mol. Cell Biol. 19, 704-713.
  3. Chae, J.H. et al. (2004) Mol. Cells 18, 271-288.
  4. Gaudillière, B. et al. (2004) Neuron 41, 229-241.
  5. Miyata, T. et al. (1999) Genes Dev. 13, 1647-1652.
  6. Naya, F.J. et al. (1997) Genes Dev. 11, 2323-2334.

Application References

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For Research Use Only. Not For Use In Diagnostic Procedures.

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