Invitrogen

NOTCH1 Monoclonal Antibody (A6)

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Questions & Answers

Questions & Answers

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NOTCH1 Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescent analysis of Notch-1 (green) showing staining in the membrane of NIH-3T3 cells. Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a Notch-1 monoclonal antibody (Product # MA5-11961) in 3% BSA-PBS at a dilution of 1:100 and incubated overnight at 4 °C in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 100x.

NOTCH1 Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescent analysis of Notch-1 (green) showing staining in the membrane of MCF-7 cells. Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a Notch-1 monoclonal antibody (Product # MA5-11961) in 3% BSA-PBS at a dilution of 1:100 and incubated overnight at 4 °C in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 100x.

NOTCH1 Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescent analysis of Notch-1 (green) showing staining in the membrane of HeLa cells. Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a Notch-1 monoclonal antibody (Product # MA5-11961) in 3% BSA-PBS at a dilution of 1:100 and incubated overnight at 4 °C in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 100x.

NOTCH1 Antibody in Western Blot (WB) — Western blot analysis of Notch-1 was performed by loading 25 µg of Hela cell lysates onto an SDS polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked at 4°C overnight. The membrane was probed with a Notch-1 monoclonal antibody (Product # MA5-11961) at a dilution of 1:200 overnight at 4°C, washed in TBST, and probed with an HRP-conjugated secondary antibody for 1 hr at room temperature in the dark. Chemiluminescent detection was performed using Pierce ECL Plus Western Blotting Substrate (Product # 32132). Results show a band at ~273 kDa.

NOTCH1 Antibody in Flow Cytometry (Flow) — Flow cytometry analysis of Notch-1 in NIH-3T3 cells (green) compared to an isotype control (blue). Cells were harvested, adjusted to a concentration of 1-5x10^6 cells/mL, fixed with 2% paraformaldehyde and washed with PBS. Cells were blocked with a 2% solution of BSA-PBS for 30 min at room temperature and incubated with a Notch-1 monoclonal antibody (Product # MA5-11961) at a dilution of 0.5 µg/test for 60 min at room temperature. Cells were then incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated secondary antibody and re-suspended in PBS for FACS analysis.

NOTCH1 Antibody in Flow Cytometry (Flow) — Flow cytometry analysis of Notch-1 in Jurkat cells (green) compared to an isotype control (blue). Cells were harvested, adjusted to a concentration of 1-5x10^6 cells/mL, fixed with 2% paraformaldehyde and washed with PBS. Cells were blocked with a 2% solution of BSA-PBS for 30 min at room temperature and incubated with a Notch-1 monoclonal antibody (Product # MA5-11961) at a dilution of 1 µg/test for 60 min at room temperature. Cells were then incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated secondary antibody and re-suspended in PBS for FACS analysis.

NOTCH1 Antibody in Western Blot (WB) — Figure 6 Epfn deficiency does not interfere with Panx3 expression. ( A ) qPCR for Panx3 and Epfn using total RNA prepared from skin of 6 weeks old Epfn +/+ and Epfn -/- mice. Results represent the mean +- SD; N = 3. * p < 0.01. NS, not significant. ( B ) Immunostaining of Panx3 (green) and Epfn (red) in si-Control or si-Epfn transfected HaCaT cells cultured in normal medium. The nuclei were counterstained with DAPI (blue in merged) ( a ). Relative fluorescence intensity comparison of images were analyzed ( b ). ( C ) Western blots of Epfn and Notch1 in Panx3 overexpressed HaCaT cells transfected with or without si-Epfn ( a ). Quantification of the ratios of Epfn/alpha-Tubulin (upper panel) and Notch1/alpha-Tubulin (lower panel). ( D ) Western blots of Epfn and Notch1 in shPanx3 transfected HaCaT cells with or without pcDNA3.1-Epfn ( a ). Quantification of the ratios of Epfn/alpha-Tubulin (upper panel) and Notch1/alpha-Tubulin (lower panel). * p < 0.01. NS, not significant.

Product Details

Applications

Tested Dilution

Publications

Western Blot (WB)

1:100-1:1,000

View 3 publications 3 publications

Immunohistochemistry (IHC)

-

View 6 publications 6 publications

Immunocytochemistry (ICC/IF)

1:20-1:200

-

Flow Cytometry (Flow)

Assay-dependent

View 3 publications 3 publications

Neutralization (Neu)

-

View 3 publications 3 publications

Product Specifications

Species Reactivity

Human, Mouse

Published species

Human, Mouse

Host/Isotype

Mouse / IgG2b

Class

Monoclonal

Type

Antibody

Clone

A6

Immunogen

Recombinant protein encoding the ligand binding region of human Notch-1

Conjugate

Unconjugated

Form

Liquid

Concentration

0.2 mg/mL

Purification

Protein A

Storage buffer

PBS, pH 7.4, with 0.2% BSA

Contains

0.09% sodium azide

Storage conditions

4° C

RRID

AB_10979079

Product Specific Information

MA5-11961 targets Notch-1 in WB, FACS and ICC/IF applications and shows reactivity with Human and mouse samples.

The MA5-11961 immunogen is recombinant protein encoding the ligand binding region of human Notch-1.

Target Information

The notch gene belongs to a family of epidermal growth factor (EGF) like homeotic genes, which encode transmembrane proteins with a variable number of cysteine-rich EGF-like repeats in the extracellular region. Four notch genes have been described in mammals: Notch1, Notch2, Notch3 and Notch4(Int-3), which have been implicated in the differentiation of the nervous system and other structures. The EGF-like proteins Delta and Serrate have been identified as ligands of Notch1. Mature Notch proteins are heterodimeric receptors derived from the cleavage of Notch pre-proteins into an extracellular subunit (NEC) containing multiple EGF-like repeats and a transmembrane subunit including intracellular region (Ntm).

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Bioinformatics

Protein Aliases: major type A protein; Motch A; mT14; Neurogenic locus notch homolog protein 1; NEXT; NICD; Notch 1; Notch 1 extracellular truncation; Notch 1 intracellular domain; Notch gene homolog 1; Notch homolog 1, translocation-associated; p300; RP23-306D20.12; RP23-306D20.12-001; Translocation-associated notch protein TAN-1; transmembrane receptor Notch1

Gene Aliases: 9930111A19Rik; AOS5; AOVD1; hN1; lin-12; Mis6; Motch; N1; NOTCH1; TAN1

UniProt ID: (Human) P46531, (Mouse) Q01705

Entrez Gene ID: (Human) 4851, (Mouse) 18128

Molecular Function: cell adhesion molecule

negative regulation of transcription from RNA polymerase II promoter in utero embryonic development cell fate specification epithelial to mesenchymal transition liver development heart looping sprouting angiogenesis positive regulation of neuroblast proliferation inflammatory response to antigenic stimulus endocardium development endocardium morphogenesis atrioventricular node development coronary vein morphogenesis aortic valve morphogenesis atrioventricular valve morphogenesis pulmonary valve morphogenesis mitral valve formation epithelial to mesenchymal transition involved in endocardial cushion formation endocardial cushion morphogenesis cardiac chamber formation cardiac ventricle morphogenesis cardiac atrium morphogenesis cardiac right atrium morphogenesis cardiac left ventricle morphogenesis cardiac right ventricle formation ventricular trabecula myocardium morphogenesis growth involved in heart morphogenesis regulation of transcription from RNA polymerase II promoter involved in myocardial precursor cell differentiation Notch signaling pathway involved in regulation of secondary heart field cardioblast proliferation cell migration involved in endocardial cushion formation pericardium morphogenesis regulation of transcription, DNA-templated transcription initiation from RNA polymerase II promoter immune response humoral immune response Notch signaling pathway positive regulation of transcription of Notch receptor target spermatogenesis determination of left/right symmetry compartment pattern specification axonogenesis foregut morphogenesis endoderm development heart development positive regulation of cell proliferation negative regulation of cell proliferation auditory receptor cell fate commitment positive regulation of epithelial to mesenchymal transition negative regulation of cell-substrate adhesion negative regulation of myotube differentiation mesenchymal cell development regulation of somitogenesis cell differentiation in spinal cord neural tube development keratinocyte differentiation negative regulation of ossification lung development positive regulation of cell migration positive regulation of BMP signaling pathway negative regulation of BMP signaling pathway forebrain development hair follicle morphogenesis organ regeneration response to corticosteroid response to muramyl dipeptide response to lipopolysaccharide embryonic hindlimb morphogenesis tube formation skeletal muscle cell differentiation cellular response to vascular endothelial growth factor stimulus tissue regeneration positive regulation of apoptotic process negative regulation of catalytic activity positive regulation of viral genome replication positive regulation of endothelial cell differentiation negative regulation of auditory receptor cell differentiation positive regulation of keratinocyte differentiation negative regulation of myoblast differentiation negative regulation of osteoblast differentiation positive regulation of Notch signaling pathway negative regulation of transcription, DNA-templated positive regulation of transcription, DNA-templated positive regulation of transcription from RNA polymerase II promoter negative regulation of calcium ion-dependent exocytosis positive regulation of JAK-STAT cascade negative regulation of photoreceptor cell differentiation somatic stem cell division astrocyte differentiation oligodendrocyte differentiation positive regulation of astrocyte differentiation negative regulation of oligodendrocyte differentiation branching morphogenesis of an epithelial tube positive regulation of viral transcription positive regulation of epithelial cell proliferation negative regulation of neurogenesis cardiac muscle tissue morphogenesis cardiac muscle cell proliferation positive regulation of cardiac muscle cell proliferation negative regulation of glial cell proliferation cilium morphogenesis cardiac epithelial to mesenchymal transition cardiac septum morphogenesis ventricular septum morphogenesis secretory columnal luminar epithelial cell differentiation involved in prostate glandular acinus development prostate gland epithelium morphogenesis regulation of epithelial cell proliferation involved in prostate gland development arterial endothelial cell differentiation venous endothelial cell differentiation cardiac vascular smooth muscle cell development endocardial cell differentiation vasculogenesis involved in coronary vascular morphogenesis coronary artery morphogenesis Notch signaling involved in heart development heart trabecula morphogenesis positive regulation of transcription from RNA polymerase II promoter in response to hypoxia left/right axis specification cellular response to follicle-stimulating hormone stimulus distal tubule development collecting duct development glomerular mesangial cell development interleukin-4 secretion negative regulation of cell migration involved in sprouting angiogenesis negative regulation of canonical Wnt signaling pathway neuronal stem cell population maintenance regulation of extracellular matrix assembly apoptotic process involved in embryonic digit morphogenesis negative regulation of stem cell differentiation negative regulation of anoikis negative regulation of pro-B cell differentiation negative regulation of endothelial cell chemotaxis angiogenesis osteoblast fate commitment regulation of cardioblast proliferation transcription, DNA-templated regulation of transcription from RNA polymerase II promoter transcription from RNA polymerase II promoter multicellular organism development epidermis development regulation of Notch signaling pathway glial cell differentiation regulation of gene expression cell differentiation neuron differentiation embryonic limb morphogenesis regulation of cell migration regulation of cell proliferation anagen cell fate commitment negative regulation of cell differentiation regulation of auditory receptor cell differentiation negative regulation of neuron differentiation positive regulation of glial cell differentiation neuron fate commitment regulation of epithelial cell proliferation regulation of neurogenesis regulation of developmental process negative regulation of cell death regulation of cell adhesion involved in heart morphogenesis positive regulation of ephrin receptor signaling pathway

Process(es)

core promoter binding transcriptional activator activity, RNA polymerase II transcription factor binding transcription factor activity, sequence-specific DNA binding enzyme inhibitor activity receptor activity Notch binding calcium ion binding protein binding enzyme binding chromatin DNA binding sequence-specific DNA binding protein heterodimerization activity chromatin binding metal ion binding