抗Notch-1 (mN1A)抗体 | Anti-Notch-1 (mN1A) Antibody

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	Genetic Strategies Validation. Knockout Validated: Notch-1 Antibody (mN1A) [NB100-78486] - Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line and Notch-1 knockout (KO) HeLa cell line. PVDF membrane was probed with 2.0 ug/mL of Mouse Anti-Human Notch-1 Monoclonal Antibody (Catalog # NB100-78486) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog #HAF018). Specific band was detected for Notch-1 at approximately 110 kDa (as indicated) in the parental HeLa cell line, but is not detectable in the knockout HeLa cell line. This experiment was conducted under reducing conditions.
	Western Blot: Notch-1 Antibody (mN1A) [NB100-78486] - Cell extracts from Jurkat (Lane 1) or mouse thymocytes (Lane 2) were Western Blot analyzed with monoclonal anti-NOTCH1 antibody. The mN1A antibody recognizes both mouse and human 270 kD full-length NOTCH1 and 110-120 kD cleaved NOTCH 1 (NICD).
	Immunocytochemistry/Immunofluorescence: Notch-1 Antibody (mN1A) [NB100-78486] - The Notch1 Antibody was tested in HEK293 cells at a 1:50 dilution against Dylight 488 (Green). Actin was counterstained against Phalloidin 568 (Red) and cells were mounted in DAPI Flouromount (Blue).
	Immunohistochemistry-Paraffin: Notch-1 Antibody (mN1A) [NB100-78486] - IHC analysis of a formalin fixed paraffin-embedded (FFPE) human pancreatic cancer using 1:10 dilution of Notch-1 antibody on a Bond Rx autostainer (Leica Biosystems). The assay involved 20 minutes of heat induced antigen retrieval (HIER) using 10mM sodium citrate buffer (pH 6.0) and endogenous peroxidase quenching with peroxide block. The sections were incubated with primary antibody for 30 minutes and Bond Polymer Refine Detection (Leica Biosystems) with DAB was used for signal development followed by counterstaining with hematoxylin. Whole slide scanning and capturing of representative images (20X) was performed using Aperio AT2 (Leica Biosystems). Cytoplasmic staining in epithelial cells was observed. Staining was performed by Histowiz.
	Flow Cytometry: Notch-1 Antibody (mN1A) [NB100-78486] - Intracellular flow cytometric staining of 1 x 10^6 CHO (A) and MCF-7 (B) cells using Notch1 antibody (dark blue). Isotype control shown in orange. An antibody concentration of 1 ug/1x10^6 cells was used.

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Background

Notch1, also known as neurogenic locus notch homolog protein 1, is a single-pass transmembrane protein that serves as the receptor for the Delta1, Jagged1 and Jagged2 ligands. Activation of Notch1 causes the cytoplasmic domain to be cleaved, releasing the notch intracellular domain (NICD) which forms a complex with RBPJ/RBPSUH and subsequently influences transcription of its target genes. Because the majority of Notch1 ligands are transmembrane proteins, Notch1 is primarily triggered by cell-to-cell contact and employs lateral inhibition mechanisms to influence local populations of cells. The Notch1 pathway plays a pivotal role in development, including cell-fate determination, proliferation, and cell contact-dependent signaling. Aberrant activation of the Notch1 pathway has been associated with numerous diseases such as bicuspid aortic valve and cancer - in particular certain leukemias.

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