mouse anti human notch1 monoclonal antibody  (Santa Cruz Biotechnology)

Journal: Scientific Reports

Article Title: Notch-1 in macrophages promoted the ischemia-reperfusion via modulating EZH2/HSF1/BRD4/SIRPα/SHP2 induced ROS and apoptosis in cardiomyocyte

doi: 10.1038/s41598-026-40683-4

Figure Lengend Snippet: Notch-1 in macrophages promotes I/R injury. ( A ) Echocardiographic measurements of left ventricular ejection fraction (LVEF), left ventricular end-diastolic diameter (LVEDD), and left ventricular end-systolic wall thickness (LVESD) in Sham, Model, and Model+Notch1-OE groups, along with statistical analysis plots for LVEF, LVEDD, and LVESD; ( B ) Histopathological changes in myocardial tissue observed via HE staining; ( C ) Immunofluorescence staining with CD68 showing experimental results in RAW264.7 cells. N = 7; Data are expressed as mean ± standard deviation; * P < 0.05; ** P < 0.01; ns P >0.05.

Article Snippet: Notch1-OE, Notch1-NC, Notch1-KD, EZH2-OE, BRD4-OE, and SHP2-KD lentiviral vector were transduced into RAW264.7 cells using transduction reagents and 8 mg/mL polybrene for 12 h. To obtain stably transduced cell lines, cells were selected in complete medium supplemented with 2.5 μg/mL puromycin (MCE, HY-B1743A) for one week post-infection.

Techniques: Staining, Immunofluorescence, Standard Deviation

Journal: JCI Insight

Article Title: Unbiased cleavage site prediction uncovers viral antagonism of host innate immunity by SARS-CoV-2 3C-like protease

doi: 10.1172/jci.insight.185739

Figure Lengend Snippet: ( A ) Western blot of in vitro cleavage of recombinant cadherin with 3CLPro. Shown are cleavage sites within the recombinant fragment, amino acid positions displayed for the full length proteins. Western blots show staining against the C-terminus (His-Tag) of each protein and 3CLPro. Recombinant proteins are a mixture of glycosylated (~90 kDa) and unglycosylated (~65 kda), corresponding to cleavage fragments of ~62 kDa and 40 kDa (respectively). For CADH6 cleavage, 2 formulations of 3CLPro were tested (3CLPro unconjugated, 3CLPro-Maltose-Binding Protein conjugated) at 2 concentrations of protease (+, 0.5 μM; ++, 1 μM). For CADH20 cleavage, only unconjugated 3CLPro digests at 1 μM concentration are shown. For 3CLPro and CADH20 staining (His-Tag), samples were run on the same gel but are noncontiguous (as indicated by line separating lanes). ( B ) Western blot of in vitro cleavage of purified human α-thrombin (IIa). Diagram shows amino acid position of unprocessed prothrombin. Position of cleavage site shown with respect to epitope of antibody used for Western blot. In total, 1 μM of purified 3CLPro was incubated with 2 μg α-thrombin overnight under reducing conditions, with or without the 3CLPro inhibitor GC376 (1 μM). ( C ) In vitro cleavage of purified recombinant NOTCH1 fragment (aa 2280–2550) with a N-terminal His-Tag. Reactions were done with 1 μM of purified 3CLPro for 1 hour. Diagram shows position of cleavage within the NOTCH1 fragment, with amino acid positions corresponding to the full-length protein. Epitope regions showed for antibody with epitope C-terminal to the cleavage site. Full-length size is ~29 kDa, with N and C-terminal fragments of 4 kDa and 25 kDa (respectively). Samples were run on the same gel but are noncontiguous. ( D ) Cellular cleavage of NOTCH1. Western blots show lysates of hIPSC cardiomyocytes expressing 3CLPro or catalytically inactive C145A variant for 48 hours. Cleavage site position within the intracellular fragment of NOTCH1 shown, as well as epitope for antibody used in Western blot.

Article Snippet: Recombinant proteins were purchased commercially: NOTCH1 (Origene, catalog TP762041), CDH6 (ACROBiosystem, catalog CA6-H5229), and CDH20 (R&D, catalog 5604-CA-050).

Techniques: Western Blot, In Vitro, Recombinant, Staining, Binding Assay, Concentration Assay, Purification, Incubation, Expressing, Variant Assay

Journal: Frontiers in Immunology

Article Title: NCF2 facilitates M2 macrophage polarization in glioblastoma through activation of the notch1–osteopontin axis

doi: 10.3389/fimmu.2026.1743950

Figure Lengend Snippet: NCF2 regulates OPN expression via Notch1 signaling in GBM. (A) Western blot analysis showing NCF2 and OPN protein expression in LN229 and T98G cells with NCF2 knockdown, and in U87 and U251 cells with NCF2 overexpression. (B) qRT-PCR analysis confirming changes in NCF2 and OPN mRNA expression in LN229 and T98G cells with NCF2 knockdown, and in U87 and U251 cells with NCF2 overexpression. qPCR was performed in triplicate for each sample. (C) Western blot analysis of NCF2, Notch1, and OPN protein expression in LN229 and T98G cells with NCF2 knockdown, with or without Notch1 agonist (N1A) treatment, and in U87 and U251 cells with NCF2 overexpression, with or without Notch1 inhibitor (N1i) treatment. (D) qRT-PCR analysis of NCF2 , Notch1 , and OPN mRNA expression in LN229 and T98G cells with NCF2 knockdown, with or without Notch1 agonist (N1A) treatment, and in U87 and U251 cells with NCF2 overexpression, with or without Notch1 inhibitor (N1i) treatment. qPCR was performed in triplicate for each sample. (“*”represents P<0.05).

Article Snippet: To investigate the involvement of the Notch1 signaling pathway, NCF2-knockdown or NCF2-overexpressing GBM cells were treated with either a Notch1 agonist (recombinant human Jagged1, 2 μg/mL; AF1277;R&D Systems) or a Notch1 inhibitor (DAPT, 10 μM; 2634; R&D Systems) for 48 h. Following treatment, the protein and mRNA expression levels of Notch1 and OPN were analyzed by western blot and qRT-PCR.

Techniques: Expressing, Western Blot, Knockdown, Over Expression, Quantitative RT-PCR

Journal: Frontiers in Oncology

Article Title: Glutamine alleviates radiation-induced intestinal injury in rats via the mTOR/Notch1 axis

doi: 10.3389/fonc.2026.1735401

Figure Lengend Snippet: Effects of Gln and RAPA on the mTOR/Notch1 axis and MUC2 expression in HT-29 cells. (A) The expression of mTOR, p-mTOR, Notch1, and GAPDH. (B) Immunofluorescence analysis of MUC2 in HT-29 cells (800×). Data are presented as mean ± SD (n = 3). One-way ANOVA was carried out followed by Tukey’s test. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

Article Snippet: To investigate the role of mTOR/notch1 pathway on Gln radio-resistance effects in colon, HT-29 cells were treated with the 10 μmol/L Jagged-1 (MCE, USA) , which activates the Notch1 receptor.

Techniques: Expressing, Immunofluorescence

Journal: Frontiers in Oncology

Article Title: Glutamine alleviates radiation-induced intestinal injury in rats via the mTOR/Notch1 axis

doi: 10.3389/fonc.2026.1735401

Figure Lengend Snippet: Effects of Gln and Jagged-1 on the mTOR/Notch1 axis and MUC2 expression in HT-29 cells. (A) The expression of mTOR, p-mTOR, Notch1, and GAPDH. (B) Immunofluorescence analysis of MUC2 in HT-29 cells (800×). Data are presented as mean ± SD (n = 3). One-way ANOVA was carried out followed by Tukey’s test. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

Article Snippet: To investigate the role of mTOR/notch1 pathway on Gln radio-resistance effects in colon, HT-29 cells were treated with the 10 μmol/L Jagged-1 (MCE, USA) , which activates the Notch1 receptor.

Techniques: Expressing, Immunofluorescence

Journal: bioRxiv

Article Title: Graded Notch Signaling Functions as a Rheostat of Lineage Plasticity and Therapy Resistance in Prostate Cancer

doi: 10.1101/2025.11.23.690056

Figure Lengend Snippet: (A-D) Box and scatter plots showing bulk RNA-sequencing data for key Notch pathway genes across a panel of human prostate cancer cell lines (LNCaP, LNCaP-abl, C4-2, C4-2B, VCaP, 22RV1, PC3, DU145, H660, LASCPC1, LNCaP_42D). (A-B) NOTCH1 and NOTCH2 Gene Expression (receptor). (C) HES1 Gene Expression (downstream effector). (D) DLL3 Gene Expression (ligand). (E) Western blot analysis confirming the protein expression of NOTCH2, DLL3, and HES1 in selected cell lines (LNCaP/AR, PC3, gTP53/RB1-R, and LASCPC1), corroborating the gene expression trends observed in the bulk RNA-seq data. Actin serves as a loading control.

Article Snippet: A pool of siRNAs targeting human NOTCH1, NOTCH2, HES1, and a non-targeting control pool were used (MedChemExpress; HY-RS09445, HY-RS09448, HY-RS06134).

Techniques: RNA Sequencing, Gene Expression, Western Blot, Expressing, Control

Journal: bioRxiv

Article Title: Graded Notch Signaling Functions as a Rheostat of Lineage Plasticity and Therapy Resistance in Prostate Cancer

doi: 10.1101/2025.11.23.690056

Figure Lengend Snippet: (A, B) Relative gene expression of Notch pathway components and lineage markers in enzalutamide-resistant cell lines, (A) gTP53/RB1-R and (B) PC3, after siRNA knockdown of specific genes ( NOTCH1, NOTCH2, HES1 ) compared to non-targeting control (NC). Significance is denoted as *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: A pool of siRNAs targeting human NOTCH1, NOTCH2, HES1, and a non-targeting control pool were used (MedChemExpress; HY-RS09445, HY-RS09448, HY-RS06134).

Techniques: Gene Expression, Knockdown, Control