Revision 13

#4373Store at -20C

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, IP, ChIP

REACTIVITY:

H M R

SENSITIVITY:

Endogenous

MW (kDa):

49

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q13562

Entrez-Gene Id:

4760

Product Information

Product Usage Information

For optimal ChIP results, use 10 μl of antibody and 10 μg of chromatin (approximately 4 x 106 cells) per IP. This antibody has been validated using SimpleChIP® Enzymatic Chromatin IP Kits.
Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:50
Chromatin IP 1:50

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Specificity / Sensitivity

NeuroD1 (D35G2) Rabbit mAb detects endogenous levels of total NeuroD1 protein.

Species Reactivity:

Human, Mouse, Rat

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide surrounding Gln15 of human NeuroD1 protein.

Background

NeuroD1 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 NeuroD1 at Ser336, which is necessary for formation and growth of dendrites (3,4). NeuroD1 is also phosphorylated at Ser274 though the results are context dependent as phosphorylation by Erk stimulates NeuroD1 activity in pancreatic β-cells while phosphorylation by GSK-3β inhibits NeuroD1 in neurons (3). NeuroD1 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 NeuroD1 become severely diabetic and die shortly after birth due to defects in β-cell differentiation (2,3,5,6). The lack of NeuroD1 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.

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

WB: Western Blotting IP: Immunoprecipitation ChIP: Chromatin IP

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

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

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Revision 13
#4373

NeuroD1 (D35G2) Rabbit mAb

Western Blotting Image 1: NeuroD1 (D35G2) Rabbit mAb Expand Image
Western blot analysis of extracts from IMR-32 cells using NeuroD1 (D35G2) Rabbit mAb.
No image available
Chromatin Immunoprecipitation Image 1: NeuroD1 (D35G2) Rabbit mAb Expand Image
Chromatin immunoprecipitations were performed with cross-linked chromatin from SH-SY5Y cells treated with Human Brain-Derived Neurotrophic Factor (BDNF) #3897 (50 ng/ml) for 30 minutes and either NeuroD1 (D35G2) Rabbit mAb or 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.