activated Notch1 antibody (ab8925)

Product Name 

activated Notch1 antibody

Product type 

Primary antibodies

Description 

Rabbit polyclonal to activated Notch1

Immunogen 

Peptide: VLLSRKRRRQHGQC (aa1755-1767), human N-terminal sequence of the cleaved N intracellular domain (NICD).
BLAST 'VLLSRKRRRQHGQC' with or

Reacts with 

Hu, Ms

Predicted to react 
(species key)

Rat
(Due to sequence homology)

Specificity 

Tested in HEK 293 cells with mouse Notch constructs (see Western blot image): Notch-1 does not react with mouse N2, N3 or N4.
Only recognises the cleaved intracellular (activated) form (NICD).

Epitope 

The epitope is only exposed after gamma secretase cleavage and is not accessible in the uncleaved form.

Tested applications 
(see key)

Flow Cyt, ICC/IF, IHC-Fr, IHC-P, WB

Abreviews 

Customer reviews (see key)

Read about Abreviews (Read all Abreviews)

Application notes 
(see key)

Recommended dilutions
Flow Cyt: Use at an assay dependent dilution (PMID 19171875).
ICC/IF: Use at an assay dependent dilution.
IHC-P: 1/200 (PMID 19147558).
IHC-Fr: Use at an assay dependent dilutio (PMID 18371421).
WB: 1/500. Detects a band of approximately 80 kDa, a cleavage product of Notch 1. Can be blocked with Notch-1 (activated) peptide (human and mouse).
There is also a contaminating band present at 100 kDa. The identity of this band is unknown.


Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.

Positive control 
(see definition)

transiently transfected 293 cells expressing recombinant myc-tagged mouse Notch constructs (see image)

Cellular localization 

Nuclear

Research areas 

Developmental Biology >> Embryogenesis >> Somatogenesis
Stem Cells >> Neural Stem Cells >> Neural Crest Stem Cells
Stem Cells >> Hematopoietic Progenitors >> Surface Molecules
Stem Cells >> Neural Stem Cells >> Surface Molecules
Stem Cells >> Neural Stem Cells >> Intracellular
Stem Cells >> Signaling Pathways >> Notch >> Nuclear
Stem Cells >> Signaling Pathways >> Notch >> Surface Molecules
Chromatin and Nuclear Signaling >> Transcription >> Other factors
Neuroscience >> Neurology process >> Notch Pathway

Relevance 

Notch family members play a role in a variety of developmental processes by controlling cell fate decisions. The Notch signaling network is an evolutionarily conserved intercellular signaling pathway which regulates interactions between physically adjacent cells. The protein is cleaved in the trans-Golgi network, and presented on the cell surface as a heterodimer. This protein functions as a receptor for membrane bound ligands. Once the Notch extracellular domain interacts with a ligand, a protease called TACE (Tumor Necrosis Factor Alpha Converting Enzyme) cleaves the Notch protein just outside the membrane. This releases the extracellular portion of Notch, which continues to interact with the ligand. The ligand plus the Notch extracellular domain is then endocytosed by the ligand expressing cell. After this first cleavage, an enzyme called gamma-secretase cleaves the remaining part of the Notch protein just inside the inner leaflet of the cell membrane. This releases the intracellular portion of the Notch protein, which then moves to the nucleus and causes various genes to be expressed. There are many other proteins involved in the intracellular portion of the Notch signalling cascade.

Notch is synthesized in the endoplasmic reticulum as an inactive form which is proteolytically cleaved by a furin-like convertase (S1 cleavage) in the trans-golgi network before it reaches the plasma membrane to yield an active, ligand-accessible form. Cleavage results in a C-terminal fragment N(TM) and a N-terminal fragment N(EC). Following ligand binding, it is cleaved (S2 cleavage) by TNF-alpha converting enzyme (TACE) to yield a membrane-associated intermediate fragment called Notch extracellular truncation (NEXT). This fragment is then cleaved by presenilin-dependent gamma-secretase (S3 cleavage) to release the intracellular domain (NICD) from the membrane.

Database links 

Raised in 

Rabbit

Clonality 

Polyclonal

Isotype 

IgG

Purity 

Whole antiserum

Storage buffer 

serum with 0.02% sodium azide

Material safety datasheet (MSDS) for this product:
Sodium Azide MSDS

Form 

Liquid

Concentration 

Concentration not determined - why is this?

Storage instructions 

Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.

See below for activated Notch1 antibody images, references, products related to ab8925 and other tools.

activated Notch1 antibody images:

References for activated Notch1 antibody (ab8925)

  This product has been used in: (two most recent references)

Radke AL et al. Mature human eosinophils express functional Notch ligands mediating eosinophil autocrine regulation. Blood 113:3092-101 (2009). EM, Flow Cyt; Human. PubMed: 19171875

Rodilla V et al. Jagged1 is the pathological link between Wnt and Notch pathways in colorectal cancer. Proc Natl Acad Sci U S A 106:6315-20 (2009). ChIP; Human. PubMed: 19325125

See all 19 publication references for this product.


If you publish research using ab8925 please let us know so that we can cite the reference on this datasheet.

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