Notch-1 Antibody (mN1A) - BSA Free

Western Blot: Notch-1 Antibody (mN1A) [NB100-78486] - 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] - 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 analyzed with monoclonal anti-NOTCH1 antibody. The mN1A antibody recognizes both mouse and human 270 kDa full-length NOTCH1 and 110-120 kDa 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 with DyLight 488 (Green). Actin was counterstained with Phalloidin 568 (Red) and cells were mounted in DAPI Flouromount (Blue).

Immunohistochemistry-Paraffin: Notch-1 Antibody (mN1A) [NB100-78486] - Analysis of 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.

• Reactivity: Hu, Mu, Rt(-)
• Applications: WB, Flow, ICC/IF, IHC, IP, CyTOF-ready, KO
• Clone: mN1A
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Format: BSA Free
• Concentration: 1.0 mg/ml

Notch-1 Antibody (mN1A) - BSA Free Summary

• Immunogen: Mouse Notch1 protein [Uniprot: Q01705]
• Specificity: Does not cross-react with Notch 2, 3, or 4.
• Isotype: IgG1 Kappa
• Clonality: Monoclonal
• Host: Mouse
• Gene: NOTCH1
• Purity: Protein G purified

Applications/Dilutions

• Dilutions:
  • CyTOF-ready
  • Flow Cytometry 1 ug per million cells
  • Immunocytochemistry/ Immunofluorescence 1:50 - 1:100
  • Immunohistochemistry 1:10 - 1:500
  • Immunohistochemistry-Paraffin 1:10 - 1:500
  • Immunoprecipitation 1:10 - 1:100
  • Knockout Validated
  • Western Blot 1 - 2 ug/mL
• Application Notes: The mN1A monoclonal antibody reacts with the intracellular domain of mouse and human Notch1 and has been reported to have highest affinity for activated intracellular Notch1 and lower affinity for full-length unprocessed/heterodimeric Notch1 forms.
  
  This antibody is CyTOF ready.
• Theoretical MW: 272.505 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• Control:

  Notch-1 Overexpression Lysate

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Reactivity Notes

Does not recognize rat Notch1.

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS
• Preservative: 0.05% Sodium Azide
• Concentration: 1.0 mg/ml
• Purity: Protein G purified

Alternate Names for Notch-1 Antibody (mN1A) - BSA Free

• EC 2.1.2.11
• EC 3.4.21.68
• hN1
• Notch (Drosophila) homolog 1 (translocation-associated)
• notch 1Notch homolog 1, translocation-associated (Drosophila)
• Notch homolog 1, translocation-associated
• Notch1
• Notch-1
• TAN1
• TAN1neurogenic locus notch homolog protein 1
• Translocation-associated notch protein TAN-1

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.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Notch-1 Antibody (NB100-78486)(10)

Publications using NB100-78486

• Alghamdi TA, Krentz NAJ, Smith N et al. Zmiz1 is required for mature B-cell function and mass expansion upon high fat feeding Molecular metabolism 2022-10-25 [PMID: 36307047] (WB, Mouse)
  
  Details:
  Dilution used in WB 1:200
• Gong M, Yan F, Yu L, Li F A dopamine-methacrylated hyaluronic acid hydrogel as an effective carrier for stem cells in skin regeneration therapy Cell death & disease 2022-08-27 [PMID: 36030275] (IHC-Fr, WB, Mouse)
  
  Details:
  WB Dilutions: 1:1000; ICC/IF Dilutions: 1:100
• Bai J, Khajavi M, Sui L et al. Angiogenic responses in a 3D micro-engineered environment of primary endothelial cells and pericytes Angiogenesis 2020-09-21 [PMID: 32955682] (ICC/IF)
• Kang H, Yang K, Xiao L et al. Osteoblast Hypoxia-Inducible Factor-1a Pathway Activation Restrains Osteoclastogenesis via the Interleukin-33-MicroRNA-34a-Notch1 Pathway. Front Immunol. 2017-10-31 [PMID: 29085370] (WB, Mouse)
• Singh NK, Kotla S, Kumar R, Rao GN Cyclic AMP Response Element Binding Protein Mediates Pathological Retinal Neovascularization via Modulating DLL4-NOTCH1 Signaling. EBioMedicine 2015-11-01 [PMID: 26870802]
• Qiu, H, Tang, X Et al. Notch1 Autoactivation via Transcriptional Regulation of Furin, Which Sustains Notch1 Signaling by Processing Notch1-Activating Proteases ADAM10 and Membrane Type 1 Matrix Metalloproteinase. Mol Cell Biol 2015-11-01 [PMID: 26283728] (IF/IHC, Mouse)
• Jundt F et al. Leukemia. 22:1587. 2008-01-01 [PMID: 18449208] (IF/IHC)
• Sun H et al. J. Cell Biol. 177:647. 2007-01-01 [PMID: 17502421] (IF/IHC)
• De Strooper B, Annaert W, Cupers P et al. A presenilin-1-dependent gamma-secretase-like protease mediates release of Notch intracellular domain. Nature. 1999-04-08 [PMID: 10206645] (WB, Mouse)
• Ray WJ, Yao M, Nowotny P et al. Evidence for a physical interaction between presenilin and Notch. Proc Natl Acad Sci U S A. 1999-03-16 [PMID: 10077672] (WB, IP, Human)

Reviews for Notch-1 Antibody (NB100-78486) (2)

Average Rating: 5

(Based on 2 reviews)

We have 2 reviews tested in 2 species: Mouse, Other.

reviewed by: Verified Customer WB Mouse 12/07/2021

Summary

• Application: Western Blot
• Sample Tested: Vascular Smooth Muscle Cells
• Species: Mouse
• Lot: NR1C2

reviewed by: Shyam Gudey WB Other 01/10/2014

Summary

• Application: Western Blot
• Species: Other
• Lot: Lot B 134275

FAQs for Notch-1 Antibody (NB100-78486). (Showing 1 - of FAQs).

Bioinformatics

• Gene Symbol: NOTCH1
• Entrez:
  • 4851(Human)
  • 18128(Mouse)
• Uniprot:
  • Q01705()