14-5785-37

antibody from Invitrogen Antibodies

Targeting: NOTCH1 TAN1

Provider product page for 14-5785-37

Western blot

Immunohistochemistry

Flow cytometry

Other assay

Antibody data

Antibody Data
Antigen structure
References [9]
Comments [0]
Validations
Other assay [3]

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Product number: 14-5785-37 - Provider product page
Provider: Invitrogen Antibodies
Product name: NOTCH1 Monoclonal Antibody (mN1A), eBioscience™
Antibody type: Monoclonal
Antigen: Other
Description: Description: The Notch family of transmembrane receptors controls cell-fate decisions during the development of many organs in a wide variety of species. After binding its ligand, the Notch receptor is cleaved in its transmembrane domain, and the resulting intracellular domain dissociates from the membrane and translocates to the nucleus, where it is able to suppress the expression of lineage-specific genes by interacting with transcriptional repressors. The mN1A antibody reacts with the intracellular domain of mouse and human Notch1, but not with Notch2, 3, or 4. The mN1A antibody has a low affinity for the full-length (unprocessed or heterodimeric cell surface) forms of Notch1. In the mouse, Notch mRNA is expressed in mouse hematopoietic cells of the fetal liver and adult thymus and bone marrow. In the thymus, Notch1 protein is detected in CD4-CD8- (double-negative) and CD4-CD8+ (single-positive) thymocytes. Studies of Notch1-transgenic cells and Notch1-null mice indicate that the receptor is involved in the regulation of lymphopoiesis and myelopoiesis. Applications Reported: This mN1A antibody has been reported for use in intracellular staining followed by flow cytometric analysis, immunoblotting (WB), and immunohistochemical staining. (Fluorochrome conjugated mN1A is recommended for use in intracellular flow cytometry.). Applications Tested: This mN1A antibody has been tested by intracellular staining and flow cytometric analysis of mouse thymocytes. This can be used at less than or equal to 1 µg per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest. Purity: Greater than 90%, as determined by SDS-PAGE. Aggregation: Less than 10%, as determined by HPLC. Filtration: 0.2 µm post-manufacturing filtered.
Reactivity: Human, Mouse
Host: Mouse
Isotype: IgG
Antibody clone number: mN1A
Vial size: 2 mg
Concentration: 0.5 mg/mL
Storage: 4° C

NOTCH1 protein structure - 14-5785-37 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Nrf2 regulates Notch signaling in HNSCC cells. A Expression of Notch1 and Notch target genes mRNA in control and Keap1- expressing SSC9 clone cells. B Expression of Notch1 and Hes1 proteins in Keap1 -mutant and Keap1- expressing SSC9 cells. C Notch1 and Hes1 mRNA and, D protein expression after Nrf2 knockdown in SSC9 cells. E Immunohistochemistry staining and expression of Nrf2 , Ki67, Notch1, and Hes1 in HNSCC clinical samples from wild-type, Nrf2 , and Keap1 mutant patients tumor tissues. F Notch1 expression in non-targeting control and Notch1 siRNA-treated SSC9 cells G Cell proliferation of SSC9 cells after knockdown of Notch1 by siRNA. H Relative mRNA expression of Hes1 and Hey1 after Notch1 knockdown in SSC9 cells. I Hes1 mRNA expression and, J Cell proliferation after knockdown of Hes1 siRNA in SSC9 cells. K Effects of Notch inhibitor DAPT and, L Assessment of cell growth after treating the cells with Notch inhibitor DAPT for 5-days. The mRNA expression levels were calculated and normalized relative to GAPDH. All experiments were run in triplicate and compared with the control group. Data presents as mean +- SEM (** P < 0.01, *** P < 0.001).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 4 Expression of NOTCH1 NICD and NOTCH-specific target gene HES1 in melanoma cell lines. (A) mRNA of NOTCH1 NICD and HES1 was measured by RT-qPCR in NHEM, WM35 and A375 and A2058 melanoma cells. Melanocytes served as the control. WM35 showed a higher level of NOTCH1 NICD mRNA and HES1 mRNA than A375 and A2058 cells. (B) Protein level of NOTCH1 NICD and HES1 was measured by western blot analysis in WM35, A375 and A2058 melanoma cell lines. All of the melanoma cell lines positively expressed NOTCH1 NICD and HES1. (B-D) Effect of TGFbeta-1 treatment on NOTCH1 NICD , HES1 mRNA and protein levels in melanoma cell lines. Treatment with rhTGF-beta1 (5 ng/ml) for 48 h induced a higher increase of NOTCH1 NICD and HES1 mRNA and their own protein levels in WM35, A375 and A2058 melanoma cells. As an internal control, GAPDH was used for normalization. Data are shown as mean +- SD of three independent experiments. The comparison of mRNA NOTCH-1 and HES1 expression in multiple groups was performed by ANOVA and Tukey's test. HES1, hairy and enhancer of split 1; RT-qPCR, reverse transcription-quantitative polymerase chain reaction; TGF-beta, transforming growth factor-beta; rh, recombinant human; GAPDH, glyceraldehyde 3-phosphate dehydrogenase. ** P

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 5 GSI effect on NOTCH1 NICD , NOTCH-specific target gene HES1 expression and on TGF-beta/Smad signaling in melanoma cell lines. (A) Inhibition of NOTCH1 NICD and NOTCH-specific target gene HES1 is illustrated after 72 h of 20 mu M GSI treatment in WM35, A375 and A2058 cells. Western blot analysis showed that GSI suppressed NOTCH1 NICD and HES1 protein levels and downregulated TGFbeta-1- induced NOTCH1 NICD , HES1 protein levels in melanoma cell lines. (B) GSI treatment consistently decreased pSMAD3 levels in all melanoma cell lines. GAPDH served as loading control. GSI, gamma-secretase inhibitor; HES1, hairy and enhancer of split 1; TGF-beta, transforming growth factor-beta; pSMAD3, phosphorylated Smad3; GAPDH, glyceraldehyde 3-phosphate dehydrogenase.