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Products:Neuroscience >> Neurology process >> Notch Pathway
Overview
Product name
Anti-activated Notch1 antibody
See all activated Notch1 products (2) ...
Description
Rabbit polyclonal to activated Notch1
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).
Tested applications
IHC-P, IHC-Fr, ICC/IF, WB, Flow Cytmore details
Cross reactivity
Reacts with
Mouse, Human
Predicted to work with
Rat
Immunogen
Peptide: VLLSRKRRRQHGQC (aa1755-1767), human N-terminal sequence of the cleaved N intracellular domain (NICD).
VLLSRKRRRQ HGQC
Epitope
The epitope is only exposed after gamma secretase cleavage and is not accessible in the uncleaved form.
Positive control
transiently transfected 293 cells expressing recombinant myc-tagged mouse Notch constructs (see image)
Properties
Form
Liquid
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.
Storage buffer
serum with 0.02% sodium azide
Purity
Whole antiserum
Primary antibody notes
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.
Clonality
Polyclonal
Isotype
IgG
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
Epigenetics and Nuclear Signaling >> Transcription >> Other factors
Neuroscience >> Neurology process >> Notch Pathway
Applications
Show applications key
Our Abpromise guarantee covers the use of ab8925 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
IHC-P: 1/200(PubMed: 19147558)
IHC-Fr: Use at an assay dependent dilution. (PubMed: 18371421)
ICC/IF: Use at an assay dependent dilution.
WB: 1/500Detects a band of approximately 80 kDa.(80kDa band is a cleavage product of Notch 1. There is also a contaminating band present at 100 kDa. The identity of this band is unknown.)
Flow Cyt: Use at an assay dependent dilution. (PubMed: 19171875)
Target
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.
Cellular localization
Nuclear
Alternative names
- hN1 antibody
- Neurogenic locus Notch homolog protein 1 antibody
- Notch homolog 1 translocation associated (Drosophila) antibody
see all
Database links
- Entrez Gene: 4851 Human
- Entrez Gene: 18128 Mouse
- Entrez Gene: 25496 Rat
- Omim: 190198 Human
- SwissProt: P46531 Human
- SwissProt: Q01705 Mouse
- SwissProt: Q07008 Rat
Anti-activated Notch1 antibody images:
Western blot - activated Notch1 antibody - ChIP Grade (ab8925)
Rabbit polyclonal (ab8925) at 1/500, against myc-tagged transiently transfected mouse Notch constructs in 293 cells.
Lane M: Mol wt markers
Lane 1 No transfection
Lane 2 N1 (mouse deleted extracellular domain)-myc
Lane 3 N1 (mouse intracellular domain)-myc
Lane 4 N2 (mouse deleted extracellular domain)-myc
Lane 5 N2 (mouse intracellular domain)-myc
Lane 6 N3 (mouse deleted extracellular domain)-myc
Lane 7 N3 (mouse intracellular domain)-myc
Lane 8 N4 (mouse deleted extracellular domain)-myc
Lane 9 N4 (mouse intracellular domain)-myc
Lane 10 N1 (mouse deleted extracellular domain)(V to G)-myc
Stacey Huppert, Ma. Xenia U. Garcia (lab of Raphael Kopan)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)-activated Notch1 antibody(ab8925)
ab8925 (1:500) diluted staining activated Notch1 in human ovarian carcinoma using an automated system (DAKO Autostainer Plus). Using this protocol there is strong staining of activated Notch 1 in nuclear/nucleolar compartments of the ovarian cortex.
Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer citrate pH6.1 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.
Immunohistochemistry (Frozen sections) - activated Notch1 antibody (ab8925)
ab8925 staining activated Notch 1 in murine dermis (including follicle) by Immunohistochemistry (Frozen sections). Tissue was fixed with paraformaldehyde and permeabilized using 0.1% Triton. Samples were then blocked with 10% serum for 1 hour at 19°C followed by incubation with the primary antibody at a 1/100 dilution for 16 hours at 4°C. An Alexa Fluor® conjugated donkey anti-rabbit polyclonal was used as secondary antibody at a 1/200 dilution. Counterstain DAPI (blue).
Image courtesy of an anonymous Abreview.
References for Anti-activated Notch1 antibody (ab8925)
This product has been referenced in:
- Zhou Det al. Mst1 and Mst2 protein kinases restrain intestinal stem cell proliferation and colonic tumorigenesis by inhibition of Yes-associated protein (Yap) overabundance. Proc Natl Acad Sci U S A : (2011).Read more (PubMed: 22042863) »
- Hao Let al. Notch-1 activates estrogen receptor-alpha-dependent transcription via IKKalpha in breast cancer cells. Oncogene 29:201-13 (2010). WB; Human.Read more (PubMed: 19838210) »
See all 22 publications for this product
Publishing research using ab8925? Please let us know so that we can cite the reference in this datasheet
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"