NOTCH3 Polyclonal antibody

• Featured Product
• KD/KO Validated

NOTCH3 Polyclonal Antibody for FC, IHC, IP, WB, ELISA

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse and More (1)

Applications

WB, IP, IHC, IF, FC, ELISA

Conjugate

Unconjugated

Publications

45

Cat no : 55114-1-AP

Synonyms

CADASIL, CASIL, Notch 3, Notch homolog 3 (Drosophila), NOTCH3

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Validation Data Gallery

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  WB analysis using 55114-1-AP

  Various lysates were subjected to SDS PAGE followed by western blot with 55114-1-AP (NOTCH3 antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.

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  WB analysis using 55114-1-AP

  Various lysates were subjected to SDS PAGE followed by western blot with 55114-1-AP (NOTCH3 antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.

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  WB analysis of COLO 320 using 55114-1-AP

  COLO 320 cells were subjected to SDS PAGE followed by western blot with 55114-1-AP (NOTCH3 antibody) at dilution of 1:500 incubated at room temperature for 1.5 hours.

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  WB analysis of U2OS using 55114-1-AP

  U2OS cells were subjected to SDS PAGE followed by western blot with 55114-1-AP (NOTCH3 antibody) at dilution of 1:600 incubated at room temperature for 1.5 hours.

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  IP experiment of HeLa using 55114-1-AP

  IP result of anti-NOTCH3 (IP:55114-1-AP, 4ug; Detection:55114-1-AP 1:800) with HeLa cells lysate 2000 ug.

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  IHC staining of human lung cancer using 55114-1-AP

  Immunohistochemical analysis of paraffin-embedded human lung cancer tissue slide using 55114-1-AP (NOTCH3 Antibody) at dilution of 1:200 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human lung cancer using 55114-1-AP

  Immunohistochemical analysis of paraffin-embedded human lung cancer tissue slide using 55114-1-AP (NOTCH3 Antibody) at dilution of 1:200 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human breast cancer using 55114-1-AP

  Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 55114-1-AP (NOTCH3 Antibody) at dilution of 1:200 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human breast cancer using 55114-1-AP

  Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 55114-1-AP (NOTCH3 Antibody) at dilution of 1:200 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human kidney using 55114-1-AP

  Immunohistochemical analysis of paraffin-embedded human kidney tissue slide using 55114-1-AP (NOTCH3 Antibody) at dilution of 1:200 (under 10x lens).

• 

  IHC staining of human kidney using 55114-1-AP

  Immunohistochemical analysis of paraffin-embedded human kidney tissue slide using 55114-1-AP (NOTCH3 Antibody) at dilution of 1:200 (under 40x lens).

• 

  FC experiment of Jurkat using 55114-1-AP

  1X10^6 Jurkat cells were intracellularly stained with 0.4 ug Anti-Human NOTCH3 (55114-1-AP) and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) at dilution 1:1000 (red), or 0.4 ug Control Antibody. Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C).

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Tested Applications

• Positive WB detected in: A2780 cells, Caco-2 cells, U2OS cells, COLO 320 cells, HeLa cells, K-562 cells
• Positive IP detected in: HeLa cells
• Positive IHC detected in: human lung cancer tissue, human breast cancer tissue, human kidney tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive FC detected in: Jurkat cells

Recommended dilution

• Western Blot (WB): WB : 1:500-1:2000
• Immunoprecipitation (IP): IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
• Immunohistochemistry (IHC): IHC : 1:50-1:500
• Flow Cytometry (FC): FC : 0.40 ug per 10^6 cells in a 100 µl suspension

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Published Applications

• KD/KO: See 4 publications below
• WB: See 36 publications below
• IHC: See 13 publications below
• IF: See 4 publications below

Product Information

55114-1-AP targets NOTCH3 in WB, IP, IHC, IF, FC, ELISA applications and shows reactivity with human, mouse samples.

• Tested Reactivity: human, mouse
• Cited Reactivity: human, mouse, rat
• Host / Isotype: Rabbit / IgG
• Class: Polyclonal
• Type: Antibody
• Immunogen: Peptide
• Full Name: Notch homolog 3 (Drosophila)
• Calculated Molecular Weight: 244 kDa
• Observed Molecular Weight: 250-260 kDa, 100 kDa
• GenBank Accession Number: NM_000435
• Gene Symbol: NOTCH3
• Gene ID (NCBI): 4854
• RRID: AB_10858393
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Antigen affinity purification
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

NOTCH3 belongs to the NOTCH family. It functions as a receptor for membrane-bound ligands Jagged1, Jagged2 and Delta1 to regulate cell-fate determination. Upon ligand activation through the released notch intracellular domain (NICD) it forms a transcriptional activator complex with RBP-J kappa and activates genes of the enhancer of split locus. NOTCH3 affects the implementation of differentiation, proliferation and apoptotic programs. Defects in NOTCH3 are the cause of cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL). The antibody is specific to NOTCH3. Notch3 is a single-pass type I membrane protein exposed at the cell surface as a heterodimer of 2321 amino acids. It is synthesized as a precursor with a molecular weight of ~280 kDa (Notch3 full-length), which is cleaved (S1 cleavage) into a 210 kDa extracellular fragment and a 97 kDa intracellular fragment (PMID: 21702048).

Publications

Species	Application	Title
mouse	WB	Cell Metab; Mitochondrial Dynamics Is Critical for the Full Pluripotency and Embryonic Developmental Potential of Pluripotent Stem Cells.; Authors - Xiuying Zhong
human,mouse,rat	WB,IF	Sci Transl Med; JAGGED-NOTCH3 signaling in vascular remodeling in pulmonary arterial hypertension.; Authors - Yu Zhang
mouse	WB	Mol Ther; METTL3-mediated m6A modification of TIMP2 mRNA promotes podocyte injury in diabetic nephropathy.; Authors - Ling Jiang
rat	WB	Biomaterials; Laminin-521 promotes quiescence in isolated stellate cells from rat liver.; Authors - Friederike Rohn
mouse	WB	Cancer Res; Manic fringe promotes a claudin-low breast cancer phenotype through notch-mediated PIK3CG induction.; Authors - Shubing Zhang
human	WB	Oncogene; Notch3 promotes prostate cancer-induced bone lesion development via MMP-3.; Authors - Sourik S Ganguly ; KD Validated

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