Revision 5

#3608Store 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, IHC-P, ChIP

REACTIVITY:

H M R

SENSITIVITY:

Endogenous

MW (kDa):

120, 300

Source/Isotype:

Rabbit IgG

UniProt ID:

#P46531

Entrez-Gene Id:

4851

Product Information

Product Usage Information

For optimal ChIP results, use 5 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
Immunohistochemistry (Paraffin) 1:200 - 1:800
Chromatin IP 1:100

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.

For a carrier free (BSA and azide free) version of this product see product #67334.

Specificity / Sensitivity

Notch1 (D1E11) XP® Rabbit mAb detects intracellular epitopes between 2400 and 2500 amino acids of human Notch1. It recognizes both the full-length (~300 KDa) and the NTM region (~120 KDa), which consists of a short extracellular juxtamembrane peptide, a transmembrane sequence and the intracellular domain (NICD). The antibody cannot detect the extracellular (ligand-binding) domain of Notch1 following cleavage at the S2 site by ADAM-type metalloproteases.

Species Reactivity:

Human, Mouse, Rat

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Pro2438 of human Notch1.

Background

Notch proteins (Notch1-4) are a family of transmembrane receptors that play important roles in development and the determination of cell fate (1). Mature Notch receptors are processed and assembled as heterodimeric proteins, with each dimer comprised of a large extracellular ligand-binding domain, a single-pass transmembrane domain, and a smaller cytoplasmic subunit (Notch intracellular domain, NICD) (2). Binding of Notch receptors to ligands of the Delta-Serrate-Lag2 (DSL) family triggers heterodimer dissociation, exposing the receptors to proteolytic cleavages; these result in release of the NICD, which translocates to the nucleus and activates transcription of downstream target genes (3,4).

  1. Artavanis-Tsakonas, S. et al. (1999) Science 284, 770-6.
  2. Chan, Y.M. and Jan, Y.N. (1998) Cell 94, 423-6.
  3. Schroeter, E.H. et al. (1998) Nature 393, 382-6.
  4. Rand, M.D. et al. (2000) Mol Cell Biol 20, 1825-35.

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 IHC-P: Immunohistochemistry (Paraffin) 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.
XP is a registered trademark of Cell Signaling Technology, Inc.
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 5
#3608

Notch1 (D1E11) XP® Rabbit mAb

Western Blotting Image 1: Notch1 (D1E11) XP® Rabbit mAb Expand Image
使用 Notch1 (D1E11) XP® Rabbit mAb 对不同细胞类型的总细胞提取物进行蛋白质印迹分析。全长 (FL) Notch 蛋白和裂解的跨膜/胞内区 (NTM) 如图所示。
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Immunohistochemistry Image 1: Notch1 (D1E11) XP® Rabbit mAb Expand Image
使用 Notch1 (D1E11) XP® Rabbit mAb 对石蜡包埋的人炎症肉芽组织进行免疫组织化学分析。
Immunohistochemistry Image 2: Notch1 (D1E11) XP® Rabbit mAb Expand Image
使用 Notch1 (D1E11) XP® Rabbit mAb 对石蜡包埋的人乳腺癌细胞进行免疫组织化学分析。
Immunohistochemistry Image 3: Notch1 (D1E11) XP® Rabbit mAb Expand Image
使用 Notch1 (D1E11) XP® Rabbit mAb 对石蜡包埋的 A2780(左图)、Jurkat(中图)和 RL(右图)细胞沉淀物进行免疫组织化学分析。A2780 和 Jurkat 均表达 Notch1,但仅 Jurkat 细胞具有裂解的 Notch1,而 RL 细胞的 Notch1 表达水平较低或不表达 Notch1。
Immunohistochemistry Image 4: Notch1 (D1E11) XP® Rabbit mAb Expand Image
使用 Notch1 (D1E11) XP® Rabbit mAb 对石蜡包埋的 MALT(黏膜相关淋巴组织)淋巴瘤周围的人胃细胞进行免疫组织化学分析。
Chromatin Immunoprecipitation Image 1: Notch1 (D1E11) XP® Rabbit mAb Expand Image
在含 γ-分泌酶抑制剂 (1 μM) 的培养基中培养 CUTLL1 细胞 3 天,随后立即收取(左小图)或洗涤并在新鲜培养基中培养 3 小时(右小图)。使用 SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003,对细胞中提取的交联染色质,在加入 Notch1 (D1E11) XP® Rabbit mAb 或 Normal Rabbit IgG #2729 后进行染色质免疫沉淀。使用人 HES1 启动子引物、SimpleChIP® Human HES4 Promoter Primers #7273 和 SimpleChIP® Human α Satellite Repeat Primers #4486,通过实时 PCR 对富集的 DNA 进行定量。将每份样品中免疫沉淀的 DNA 的量表现为相对于所输入染色质总量(等于 1)的信号。