Revision 1

#62953Store 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, 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 Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:50
Immunofluorescence (Frozen) 1:1600
Immunofluorescence (Immunocytochemistry) 1:400 - 1:1600

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 (E3E4F) Rabbit mAb recognizes endogenous levels of total NeuroD1 protein.

Species Reactivity:

Human, Mouse, Rat

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding His341 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 IF-F: Immunofluorescence (Frozen) IF-IC: Immunofluorescence (Immunocytochemistry)

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.
KARPAS cell line source: Dr. Abraham Karpas at the University of Cambridge.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

限制使用

除非 CST 的合法授书代表以书面形式书行明确同意,否书以下条款适用于 CST、其关书方或分书商提供的书品。 任何书充本条款或与本条款不同的客书条款和条件,除非书 CST 的合法授书代表以书面形式书独接受, 否书均被拒书,并且无效。

专品专有“专供研究使用”的专专或专似的专专声明, 且未专得美国食品和专品管理局或其他外国或国内专管机专专专任何用途的批准、准专或专可。客专不得将任何专品用于任何专断或治专目的, 或以任何不符合专专声明的方式使用专品。CST 专售或专可的专品提供专作专最专用专的客专,且专用于研专用途。将专品用于专断、专防或治专目的, 或专专售(专独或作专专成)或其他商专目的而专专专品,均需要 CST 的专独专可。客专:(a) 不得专独或与其他材料专合向任何第三方出售、专可、 出借、捐专或以其他方式专专或提供任何专品,或使用专品制造任何商专专品,(b) 不得复制、修改、逆向工程、反专专、 反专专专品或以其他方式专专专专专品的基专专专或技专,或使用专品开专任何与 CST 的专品或服专专争的专品或服专, (c) 不得更改或专除专品上的任何商专、商品名称、徽专、专利或版专声明或专专,(d) 只能根据 CST 的专品专售条款和任何适用文档使用专品, (e) 专遵守客专与专品一起使用的任何第三方专品或服专的任何专可、服专条款或专似专专

Revision 1
#62953

NeuroD1 (E3E4F) Rabbit mAb

Western Blotting Image 1: NeuroD1 (E3E4F) Rabbit mAb Expand Image
使用 NeuroD1 (E3E4F) Rabbit mAb(上图)或 GAPDH (D16H11) XP® Rabbit mAb #5174(下图)对来自各种组织的提取物进行蛋白质印迹分析。
Western Blotting Image 2: NeuroD1 (E3E4F) Rabbit mAb Expand Image
使用 NeuroD1 (E3E4F) Rabbit mAb(上图)或 GAPDH (D16H11) XP® Rabbit mAb #5174(下图)对来自各种细胞系的提取物进行蛋白质印迹分析。
Immunoprecipitation Image 1: NeuroD1 (E3E4F) Rabbit mAb Expand Image
对来自 IMR-32 细胞提取物的 NeuroD1 蛋白进行免疫沉淀。泳道 1 为 10% 输入,泳道 2 为 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900,泳道 3 为 NeuroD1 (E3E4F) Rabbit mAb。使用 NeuroD1 (D90G12) Rabbit mAb #7019 进行蛋白质印迹分析。Mouse Anti-Rabbit IgG (Light-Chain Specific) (D4W3E) mAb (HRP Conjugate) #93702 用作二抗。
Immunofluorescence Image 1: NeuroD1 (E3E4F) Rabbit mAb Expand Image
对使用 NeuroD1 (E3E4F) Rabbit mAb(左图,绿色)标记的固定冷冻的小鼠胰腺组织进行共聚焦免疫荧光分析。然后将游离的二抗结合位点用 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 封闭,之后用 Ras (E4K9L) Rabbit mAb (Alexa Fluor® 555 Conjugate) #64520(右图,伪彩灰色)、Caveolin-1 (D46G3) XP® Rabbit mAb (Alexa Fluor® 647 Conjugate) #31411(右图,红色)和 ProLong Gold Antifade Reagent with DAPI #8961(右图,蓝色)共标记。
Immunofluorescence Image 2: NeuroD1 (E3E4F) Rabbit mAb Expand Image
对使用 NeuroD1 (E3E4F) Rabbit mAb(左图,绿色)标记以及使用 β3-Tubulin (E9F3E) Mouse mAb #45058(右图,红色)和 ProLong Gold Antifade Reagent with DAPI #8961(右图,蓝色)共标记的固定冷冻的小鼠视网膜组织进行共聚焦免疫荧光分析。
Immunofluorescence Image 3: NeuroD1 (E3E4F) Rabbit mAb Expand Image
对使用 NeuroD1 (E3E4F) Rabbit mAb(左图,绿色)标记的固定冰冻的小鼠小脑组织进行共聚焦免疫荧光分析。然后将游离的二抗结合位点用 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 封闭,之后用 Myelin Basic Protein (D8X4Q) XP® Rabbit mAb (Alexa Fluor® 555 Conjugate) #84987(右图,伪彩灰色)和 β3-Tubulin (E9F3E) Mouse mAb #45058(右图,红色)共标记。
Immunofluorescence Image 4: NeuroD1 (E3E4F) Rabbit mAb Expand Image
对使用 NeuroD1 (E3E4F) Rabbit mAb(左图,绿色)标记的固定冷冻的小鼠海马组织进行共聚焦免疫荧光分析。然后将游离的二抗结合位点用 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 封闭,之后用 Myelin Basic Protein (D8X4Q) XP® Rabbit mAb (Alexa Fluor® 555 Conjugate) #84987(右图,伪彩灰色)、β3-Tubulin (E9F3E) Mouse mAb #45058(右图,红色)和 ProLong Gold Antifade Reagent with DAPI #8961(右图,蓝色)共标记。
Immunofluorescence Image 1: NeuroD1 (E3E4F) Rabbit mAb Expand Image
使用 NeuroD1 (E3E4F) Rabbit mAb(绿色)、DyLight 554 Phalloidin #13054(红色)和 DAPI #4083(蓝色)对 WERI-Rb-1 细胞(左图,阳性)和 KARPAS 299 细胞(右图,阴性)进行共聚焦免疫荧光分析。