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Anti-activated Notch1 antibody (ab8925)

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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

  • Western blot - activated Notch1 antibody - ChIP Grade (ab8925)Western blot - activated Notch1 antibody - ChIP Grade (ab8925) image (enlarge)

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)-activated Notch1 antibody(ab8925)Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)-activated Notch1 antibody(ab8925) image (enlarge)

  • Immunohistochemistry (Frozen sections) - activated Notch1 antibody (ab8925)Immunohistochemistry (Frozen sections) - activated Notch1 antibody (ab8925) image (enlarge)

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.

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
  • NOTCH1 antibody
  • TAN1 antibody
  • Translocation-associated Notch protein TAN-1 antibody
see all

Anti-activated Notch1 antibody images:

  Western blot - activated Notch1 antibody - ChIP Grade (ab8925)

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)

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)

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.

See 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"