anti notch2 nicd  (Cell Signaling Technology Inc)

Journal: The American Journal of Pathology

Article Title: The Hajdu Cheney Mutation Is a Determinant of B-Cell Allocation of the Splenic Marginal Zone

doi: 10.1016/j.ajpath.2017.09.010

Figure Lengend Snippet: Primers Used for RT-qPCR Determinations

Article Snippet: To detect the Notch2 NICD, a hybridoma-produced monoclonal antibody against Notch2 (c651.6DbHN; Developmental Studies Hybridoma Bank, University of Iowa, Iowa City, IA), generated with an epitope coded by bp 6508 to 6906 of Notch2 , was used.

Techniques: Sequencing

Journal: The American Journal of Pathology

Article Title: The Hajdu Cheney Mutation Is a Determinant of B-Cell Allocation of the Splenic Marginal Zone

doi: 10.1016/j.ajpath.2017.09.010

Figure Lengend Snippet: Expression of Notch2 target genes and mitogen-activated protein kinase and phosphatidylinositol 3 kinase (PI3K)/Akt signaling pathway elements in Notch2HCS and littermate control mice. A: Total RNA from CD19+ cells was extracted and gene expression determined by quantitative RT-PCR. B and C: Western blot analysis of total cell lysates (100 μg) from spleen cells was fractionated by SDS-PAGE, and blots were probed in B with anti-Notch2 antibodies to detect the Notch2 intracellular domain (N2ICD) and in C with anti–phospho-Akt (pAkt) Ser473 and anti–phospho-Jun N-terminal kinase (pJNK) antibodies. Blots were stripped and reprobed with anti-Akt, anti-JNK, or anti–β-actin antibodies to verify protein loading. The ratios of pAkt/Akt to pJNK/JNK were quantified by Odyssey Infrared Imaging Systems software version 2.1 (LI-COR Biosciences, Lincoln, NE) and are shown under each blot. Molecular weight (Mr) ladder is shown along the left. Data are expressed as mean (95% CI) copy numbers of Notch26955T>C mutant, Hes5, and Hes1, corrected for Rpl38 (A). n = 4 biological replicates (A). ∗P < 0.05 between Notch2HCS mutant and wild-type control cells (A).

Article Snippet: To detect the Notch2 NICD, a hybridoma-produced monoclonal antibody against Notch2 (c651.6DbHN; Developmental Studies Hybridoma Bank, University of Iowa, Iowa City, IA), generated with an epitope coded by bp 6508 to 6906 of Notch2 , was used.

Techniques: Expressing, Control, Gene Expression, Quantitative RT-PCR, Western Blot, SDS Page, Imaging, Software, Molecular Weight, Mutagenesis

Journal: The American Journal of Pathology

Article Title: The Hajdu Cheney Mutation Is a Determinant of B-Cell Allocation of the Splenic Marginal Zone

doi: 10.1016/j.ajpath.2017.09.010

Figure Lengend Snippet: B-Cell Populations in the Bone Marrow and Spleen of 2-Month–Old Notch2HCS Mutant Male Mice and Sex-Matched Littermate Controls Treated with Anti-Notch2 NRR or Anti-Ragweed Control Antibody, Both at 10 mg/kg, for 4 Weeks

Article Snippet: To detect the Notch2 NICD, a hybridoma-produced monoclonal antibody against Notch2 (c651.6DbHN; Developmental Studies Hybridoma Bank, University of Iowa, Iowa City, IA), generated with an epitope coded by bp 6508 to 6906 of Notch2 , was used.

Techniques: Mutagenesis, Control, Clinical Proteomics

Journal: The American Journal of Pathology

Article Title: The Hajdu Cheney Mutation Is a Determinant of B-Cell Allocation of the Splenic Marginal Zone

doi: 10.1016/j.ajpath.2017.09.010

Figure Lengend Snippet: Distribution of marginal zone B cells (MZBs) assessed by flow cytometry of spleen cells isolated from 2-month–old Notch2HCS mutant male mice and sex-matched littermate controls treated with anti–Notch2 negative regulatory region (NRR) antibody or anti-ragweed control antibody, both at 10 mg/kg for 4 weeks before sacrifice. A: Representative dot plots of spleen cells from 2-month–old control mice and Notch2HCS mice treated with anti-ragweed antibodies or anti-Notch2 NRR antibodies. Cells were stained with anti-CD21/35 and anti-CD23 antibodies. Circled areas define CD21/35highCD23− MZBs and CD21/35intCD23+ follicular B cells gated on B220+/CD19+IgM+ B-cell populations. B: Frequencies of MZBs and follicular B cells (FoBs) in Notch2HCS mutant and wild-type littermates treated with anti-ragweed or anti-Notch2 NRR antibodies. Data are expressed as mean (95% CI) frequencies of MZBs and FoBs (A); data are expressed as means (95% CI) (B). n = 4 biological replicates. ∗P < 0.05 Notch2HCS versus control; †P < 0.05 anti-ragweed antibody–treated versus anti-NRR antibody–wild-type mice; ‡P < 0.05 anti-ragweed antibody–treated versus anti-Notch2 NRR antibody–Notch2HCS mice.

Article Snippet: To detect the Notch2 NICD, a hybridoma-produced monoclonal antibody against Notch2 (c651.6DbHN; Developmental Studies Hybridoma Bank, University of Iowa, Iowa City, IA), generated with an epitope coded by bp 6508 to 6906 of Notch2 , was used.

Techniques: Flow Cytometry, Isolation, Mutagenesis, Control, Staining