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Notch Receptor Interaction Antibody Sampler Kit #8658

Kit Includes	Quantity	Applications	Reactivity	MW (kDa)	Isotype
ADAM9 (D64B5) Rabbit mAb #4151	40 µl	W	H M R Mk	100-115, 75-80	Rabbit IgG
DLL1 Antibody #2588	40 µl	W IP	R (H)	82	Rabbit
DLL3 (G93) Antibody #2483	40 µl	W IP	R (M)	65	Rabbit
DLL4 Antibody #2589	40 µl	W IP	H	75-80 kDa	Rabbit
Jagged1 (28H8) Rabbit mAb #2620	40 µl	W IP	H M (R)	180	Rabbit IgG
Jagged2 (C23D2) Rabbit mAb #2210	40 µl	W IP	H R	150 kDa	Rabbit IgG
Numb (C29G11) Rabbit mAb #2756	40 µl	W IP IF-IC F	H M R Mk	72, 74	Rabbit IgG
RBPSUH (D10A4) XP® Rabbit mAb #5313	40 µl	W ChIP	H M R Mk	61	Rabbit IgG
TACE (D22H4) Rabbit mAb #6978	40 µl	W	H	135	Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody #7074	100 µl				Goat

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry  ChIP=Chromatin IP
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Specificity / Sensitivity

ADAM9 (D64B5) Rabbit mAb, DLL4 Antibody, Jagged1 (28H8) Rabbit mAb, Jagged2 (C23D2) Rabbit mAb, Numb (C29G11) Rabbit mAb, RBPSUH (D10A4) XP® Rabbit mAb, and TACE (D22H4) Rabbit mAb recognize endogenous levels of total respecitive protein. DLL1 Antibody recognizes only transfected levels of DLL1 protein. It does not recognize transfected levels of rat DLL3 or human DLL4. DLL3 (G93) Antibody recognizes only transfected levels of DLL3 protein. It does not recognize transfected levels of rat DLL1 or human DLL4. Jagged1 (28H8) Rabbit mAb does not cross-react with Jagged2. Jagged2 (C23D2) Rabbit mAb does not cross-react with Jagged1.

Description

The Notch Receptor Interaction Antibody Sampler Kit provides an economical means to evaluate Notch signaling. The kit contains enough primary antibody to perform four western blots per primary.

Source / Purification

Monoclonal antibodies are produced by immunizing animals either with a recombinant protein specific to the amino terminus of human TACE protein or with a synthetic peptide corresponding to residues surrounding Glu1140 (intracellular region) of human Jagged1 protein, residues surrounding Ala117 of human Jagged2 protein, residues surrounding Ala570 of human Numb protein, or residues near the carboxy terminus of human ADAM9 protein or residues surrounding Gln110 of human RBPSUH protein.Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ala627 of human DLL1 protein, residues surrounding Gly93 of mouse DLL3 protein, residues surrounding Leu617 of human DLL4 protein. Polyclonal antibodies are purified by protein A and peptide affinity chromatography.

Background

Notch signaling is activated upon engagement of the Notch receptor with its ligands, the Delta, Serrate, Lag2 (DSL) single-pass type I membrane proteins. DSL proteins contain multiple EGF-like repeats and a DSL domain that is required for binding to Notch (1,2). Five DSL proteins have been identified in mammals: Jagged1, Jagged2, Delta-like (DLL) 1, 3, and 4 (3). Ligand binding to the Notch receptor results in two sequential proteolytic cleavages of the receptor by the ADAM protease and the γ-secretase complex. The intracellular domain of Notch is released and then translocates to the nucleus where it activates transcription. Notch ligands may also be processed in a similiar manner, suggesting bi-directional signaling through receptor-ligand interactions (4-6).TNF-α converting enzyme (TACE), also known as ADAM17, is a transmembrane metalloprotease that plays a key role in the cleavage of a number cell surface molecules in a process known as “shedding". TACE is abundantly expressed in many adult tissues, but in fetal development, expression is differentially regulated (7). TACE activates Notch in a ligand-independent manner and has been shown to play a role in the development of the Drosophila nervous system (8).Recombining Binding Protein, SUppressor of Hairless (RBPSUH), also termed RBP-J or CSL, is the DNA-binding component of the transcription complex regulated by canonical Notch signaling. In the absence of Notch activation, RBPSUH suppresses target gene expression through interactions with a co-repressor complex containing histone deacetylase. Upon activation of Notch receptors, the Notch intracellular domain (NICD) translocates to the nucleus and binds to RBPSUH. This displaces the co-repressor complex and replaces it with a transcription activation complex that includes Mastermind-like (MAML) proteins and histone acetylase p300, leading to transcriptional activation of Notch target genes (9-11).Numb contains an amino-terminal phosphotyrosine-binding (PTB) domain and carboxy-terminal endocytic binding motifs for α-adaptin and EH (Eps15 homology) domain-containing proteins, indicating a role in endocytosis (12,13). There are four mammalian Numb splicing isoforms that are differentially expressed and may have distinct functions (14-16). Numb acts as a negative regulator of Notch signaling by promoting ubiquitination and degradation of Notch (17). The protein is asymmetrically segregated into one daughter cell during cell division, producing two daughter cells with different responses to Notch signaling and different cell fates (18,19).

1. Wilson, A. and Radtke, F. (2006) FEBS Lett 580, 2860-8.
2. Hansson, E.M. et al. (2004) Semin Cancer Biol 14, 320-8.
3. Chiba, S. (2006) Stem Cells 24, 2437-47.
4. Bland, C.E. et al. (2003) J Biol Chem 278, 13607-10.
5. Six, E. et al. (2003) Proc Natl Acad Sci U S A 100, 7638-43.
6. LaVoie, M.J. and Selkoe, D.J. (2003) J Biol Chem 278, 34427-37.
7. Black, R.A. et al. (1997) Nature 385, 729-33.
8. Delwig, A. and Rand, M.D. (2008) Cell Mol Life Sci 65, 2232-43.
9. Ehebauer, M. et al. (2006) Sci STKE 2006, cm7.
10. Borggrefe, T. and Oswald, F. (2009) Cell Mol Life Sci 66, 1631-46.
11. Kopan, R. and Ilagan, M.X. (2009) Cell 137, 216-33.
12. Berdnik, D. et al. (2002) Dev Cell 3, 221-31.
13. Santolini, E. et al. (2000) J Cell Biol 151, 1345-52.
14. Dho, S.E. et al. (1999) J Biol Chem 274, 33097-104.
15. Verdi, J.M. et al. (1999) Proc Natl Acad Sci U S A 96, 10472-6.
16. Verdi, J.M. et al. (1999) Proc Natl Acad Sci U S A 96, 10472-6.
17. McGill, M.A. and McGlade, C.J. (2003) J Biol Chem 278, 23196-203.
18. Verdi, J.M. et al. (1996) Curr Biol 6, 1134-45.
19. Reugels, A.M. et al. (2006) Dev Dyn 235, 934-48.

Application References

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!

Protocols

2210:

Immunoprecipitation, Western Blotting

2483:

Immunoprecipitation, Western Blotting

2588:

Immunoprecipitation, Western Blotting

2589:

Immunoprecipitation, Western Blotting

2620:

Immunoprecipitation, Western Blotting

2756:

Flow, Immunofluorescence, Immunoprecipitation, Western Blotting

4151:

Western Blotting

5313:

ChIP Agarose, ChIP Magnetic, Western Blotting

6978:

Western Blotting

Companion Products

• 7071 Phototope^®-HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked Antibody
• 7003 20X LumiGLO^® Reagent and 20X Peroxide
• 9998 BSA
• 9999 Nonfat Dry Milk
• 9997 Tris Buffered Saline with Tween 20 (TBST-10X)
• 7720 Prestained Protein Marker, Broad Range (Premixed Format)
• 7727 Biotinylated Protein Ladder Detection Pack
• 7722 Blue Loading Buffer Pack
• 7723 Red Loading Buffer Pack
• 7074 Anti-rabbit IgG, HRP-linked Antibody
• 7075 Anti-biotin, HRP-linked Antibody

Selected rabbit monoclonal antibodies are produced under license (granting certain rights including those under U. S. Patents No. 5,675,063 and in some instances 7,429,487) from Epitomics, Inc. 

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For Research Use Only. Not For Use In Diagnostic Procedures.