Name: Notch Receptor Interaction Antibody Sampler Kit
Brand: Cell Signaling Technology
Price: 650.00 USD
Availability: InStock

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Notch Receptor Interaction Antibody Sampler Kit: Image 1 — Western blot analysis of total cell lysates from HeLa, SK-OV-3 and SR cells using Jagged2 (C23D2) Rabbit mAb.

Notch Receptor Interaction Antibody Sampler Kit: Image 2 — Western blot analysis of COS cell extracts, untransfected or transiently transfected with a construct expressing rat DLL3 protein, using DLL3 (G93) Antibody.

Notch Receptor Interaction Antibody Sampler Kit: Image 3 — Western blot analysis of extracts from COS cells, untransfected or transiently transfected with a construct expressing rat DLL1 protein, using DLL1 Antibody.

Notch Receptor Interaction Antibody Sampler Kit: Image 4 — Western blot analysis of extracts from HUVEC and COS cells, untransfected or transiently transfected with a construct expressing human DLL4, using DLL4 Antibody.

Notch Receptor Interaction Antibody Sampler Kit: Image 5 — Western blot analysis of total cell lysates from HepG2 and LNCaP cells, using Jagged1 (28H8) Rabbit mAb.

Notch Receptor Interaction Antibody Sampler Kit: Image 6 — Western blot analysis of extracts from various cell lines using Numb (C29G11) Rabbit mAb.

Notch Receptor Interaction Antibody Sampler Kit: Image 7 — Western blot analysis of extracts from various cell types using ADAM9 (D64B5) Rabbit mAb.

Notch Receptor Interaction Antibody Sampler Kit: Image 8 — Western blot analysis of extracts from various cell lines using RBPSUH (D10A4) XP^® Rabbit mAb.

Notch Receptor Interaction Antibody Sampler Kit: Image 9 — Western blot analysis of extracts from Raji and Jurkat cells, untreated or treated with peptide N-glycosidase F (PNGase F), using TACE (D22H4) Rabbit mAb.

Notch Receptor Interaction Antibody Sampler Kit: Image 10 — After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO^® is added and emits light during enzyme catalyzed decomposition.

Notch Receptor Interaction Antibody Sampler Kit: Image 11 — Immunoprecipitation/western blot analysis of lysates from HeLa cells. Lane 1 contains lysate input (10%), lane 2 was immunoprecipitated with non-specific rabbit IgG (#3900), lane 3 was immunoprecipitated with Numb (C29G11) Rabbit mAb #2756. Western blot analysis was performed using Numb (C29G11) Rabbit mAb #2756.

Notch Receptor Interaction Antibody Sampler Kit: Image 12 — Immunohistochemical analysis of paraffin-embedded human colon carcinoma using RBPSUH (D10A4) XP^® Rabbit mAb.

Notch Receptor Interaction Antibody Sampler Kit: Image 13 — Confocal immunofluorescent analysis of HeLa cells using Numb (C29G11) Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5^® #4084 (fluorescent DNA dye).

Notch Receptor Interaction Antibody Sampler Kit: Image 14 — Immunohistochemical analysis of paraffin-embedded human lung carcinoma using RBPSUH (D10A4) XP^® Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).

Notch Receptor Interaction Antibody Sampler Kit: Image 15 — Flow cytometric analysis of HeLa cells using Numb (C29G11) Rabbit mAb (green solid line) compared to concentration-matched Rabbit (DA1E) mAb IgG XP^® Isotype Control #3900 (green dashed line). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor^® 488 Conjugate) #4412 was used as a secondary antibody.

Notch Receptor Interaction Antibody Sampler Kit: Image 16 — Immunohistochemical analysis of paraffin-embedded E18.5 mouse lung, Rbpjk ^F/+ Shh^+/+ (wild type, left) or Rbpjk ^F/- Shh^cre/+ (Rbpjk conditional knock out, right), using RBPSUH (D10A4) XP^® Rabbit mAb. Note lack of staining in the bronchial epithelial cells in the conditional knock out tissue (right). Tissue courtesy of Dr. Wellington Cardosa, Boston University School of Medicine.

Notch Receptor Interaction Antibody Sampler Kit: Image 17 — Immunohistochemical analysis of paraffin-embedded human lung carcinoma using RBPSUH (D10A4) XP^® Rabbit mAb.

Notch Receptor Interaction Antibody Sampler Kit: Image 18 — Immunohistochemical analysis of paraffin-embedded mouse lymph node using RBPSUH (D10A4) XP^® Rabbit mAb.

Notch Receptor Interaction Antibody Sampler Kit: Image 19 — CUTLL1 cells were cultured in media with γ-secretase inhibitor (1 μM, 3 d) and then either harvested immediately (left panel) or washed and cultured in fresh media for 3 h (right panel). Chromatin immunoprecipitations were performed with cross-linked chromatin from cells and RBPSUH (D10A4) XP^® Rabbit mAb or 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.

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To Purchase # 8658

Cat. #	Size	Qty.	Price
8658T	1 Kit (9 x 20 microliters)		$ 650

Product Includes
Related Products

Product Includes	Quantity	Applications	Reactivity	MW(kDa)	Isotype
ADAM9 (D64B5) Rabbit mAb 4151	20 µl	WB	H M R Mk	100-115, 75-80	Rabbit IgG
DLL1 Antibody 2588	20 µl	WB	R	82	Rabbit
DLL3 (G93) Antibody 2483	20 µl	WB IP	R	65	Rabbit
DLL4 Antibody 2589	20 µl	WB IP	H	75-80	Rabbit
Jagged1 (28H8) Rabbit mAb 2620	20 µl	WB IP	H M	180	Rabbit IgG
Jagged2 (C23D2) Rabbit mAb 2210	20 µl	WB IP	H R	150	Rabbit IgG
Numb (C29G11) Rabbit mAb 2756	20 µl	WB IP IF F	H M R Mk	72, 74	Rabbit IgG
RBPSUH (D10A4) XP^® Rabbit mAb 5313	20 µl	WB IHC ChIP	H M R Mk	61	Rabbit IgG
TACE (D22H4) Rabbit mAb 6978	20 µl	WB	H	135	Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074	100 µl	WB			Goat

Kit Usage Information

Protocols

2210: Western Blotting, Immunoprecipitation (Magnetic)
2483: Western Blotting, Immunoprecipitation (Magnetic)
2588: Western Blotting
2589: Western Blotting, Immunoprecipitation (Magnetic)
2620: Western Blotting, Immunoprecipitation (Magnetic)
2756: Western Blotting, Immunoprecipitation (Agarose), Immunofluorescence, Flow
4151: Western Blotting
5313: Western Blotting, Immunohistochemistry (Paraffin), ChIP Magnetic
6978: Western Blotting
7074: Western Blotting

Product Description

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

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.

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).

Pathways & Proteins

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Database Links

UniProt ID: Q13443 , Q9NR61 , Q9Y219 , P78504 , O00548 , Q06330 , Q9NYJ7 , P78536 , P49757

Entrez-Gene Id: 8754 , 54567 , 3714 , 182 , 28514 , 3516 , 10683 , 6868 , 8650

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Limited Uses

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

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.

Notch Receptor Interaction Antibody Sampler Kit #8658