Cell Signaling Technology

Product Pathways - Development

Cleaved Notch1 (Val1744) (D3B8) Rabbit mAb #4147

NICD   Notch  

No. Size Price
4147S 100 µl ( 10 western blots ) ¥3,900.00 现货查询 购买询价
4147T 20 µl ( 2 western blots ) ¥1,500.00 现货查询 购买询价
4147 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human,Mouse,Rat, Endogenous 110 Rabbit IgG
IP 1:200
ChIP 1:200

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IP=Immunoprecipitation, ChIP=Chromatin IP,

Specificity / Sensitivity

Cleaved Notch1 (Val1744) (D3B8) Rabbit mAb detects endogenous levels of the cytosolic domain of Notch1 only when cleaved between Gly1743 and Val1744. The antibody does not recognize full length Notch1 or Notch1 cleaved at other positions. It detects Cleaved Notch1 with different sizes in different cell lines due to mutations at the distant C-termini of Notch1 (see refs 6 and 7).

剪切的Notch1 (Val1744) (D3B8)Rabbit mAb兔单抗只识别甘氨酸1743和缬氨酸1744倍剪切的内源性Notch1 蛋白的胞质结构域。此抗体不识别全长Notch1或Notch1其他位置剪切型Notch1。不同细胞系会存在Notch1远端羧基末端的突变,因此导致抗体检测不同大小的剪切型Notch1(见参考文献6和7).

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to the sequence at the Val1744 cleavage site of human Notch1.


Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Cleaved Notch1 (Val1744) (D3B8) Rabbit mAb (upper) or Notch1 (D1E11) XP® Rabbit mAb #3608 (lower).

使用Cleaved Notch1 (Val1744) (D3B8) Rabbit mAb(上) or Notch1 (D1E11) XP® mAb#3608 (下)对多种细胞提取物进行western blot分析。

Chromatin IP

Chromatin IP

CUTLL1 cells were cultured in media with γ-secretase inhibitor (1μM) for 3 days and then either harvested immediately (left panel) or washed and cultured in fresh media for 3h (right panel). Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 cells and 2.5 µl of Cleaved Notch1 (Val1744) (D3B8) 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 human HES1 promoter primers, SimpleChIP® Human HES4 Promoter Primers #7273, 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.

CUTLL1细胞在包含γ-分泌酶抑制剂(1 μM) 的培养基中培养三天后,直接收取细胞(左)或清洗后在新鲜培养基中培养3小时。使用SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003将4 x 106 处理的CUTLL1细胞与2.5 μl的Cleaved Notch1 (Val1744) (D3B8) Rabbit mAb或2μl的Normal Rabbit IgG #2729交联然后进行染色质免疫共沉淀实验。富集的DNA使用SimpleChIP® Human HES1 Promoter 引物,#7273 human HES4 promoter 引物,和SimpleChIP® Human α Satellite Repeat 引物 #4486以real-time PCR的方式定量。各样品沉淀得到的DNA量相对于input染色质总量进行相对定量,Input中染色质量设定值为1。


Notch1 is a transmembrane protein functioning in development and the determination of cell fate (1). During maturation, the notch molecule is cleaved by a furin-like convertase at its extracellular domain (2). Upon binding to a ligand such as Delta1, or upon extracellular calcium depletion, the carboxy-terminal notch1 fragment is released and further cleaved between Gly1743 and Val1744 (3,4). The resulting activated cytosolic fragment translocates to the nucleus where it activates transcription.

Notch1是一个在细胞发育和命运决定过程中发挥功能的跨膜蛋白(1)。在notch1成熟过程中,notch1分子细胞外结构域会被一个成对碱性氨基酸蛋白酶样的转化酶剪切(2)。在与配体如Delta1结合或是细胞外钙离子耗竭后,notch1的羧基端被释放并随后在Gly1743 和 Val1744之间被切开(3,4)。结果导致细胞内处于活化状态的notch1片段转运至细胞核内并转录激活。

Constitutively activated Notch1 signaling is associated with the majority of cases of T cell acute lymphoblastic leukemia (T-ALL). The activation is either due to mutations in Notch1 itself or in the components of ubiquitin ligase complex, namely FBW7 (5-7).


  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.
  5. Weng, A.P. et al. (2004) Science 306, 269-71.
  6. Thompson, B.J. et al. (2007) J Exp Med 204, 1825-35.

Application References

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