Western blot analysis of NOTCH 1 was performed by loading 30ug of HEK293T whole cell lysate per well onto an SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% non-fat milk in TBST for 1 hour at room temperature. The membrane was probed with a NOTCH 1 monoclonal antibody (Product # MA1-81888) at a dilution of 1:500 overnight at 4°C, washed in TBST, and probed with an HRP-conjugated goat anti-mouse IgG secondary antibody at a dilution of 1:40,000 for 1 hour at room temperature. Chemiluminescent detection was performed using ECL substrate. Data courtesy of the Innovators Program.
|Tested species reactivity||Human, Mouse|
|Published species reactivity||Human|
|Host / Isotype||Mouse / IgG1|
|Immunogen||Synthetic peptide corresponding to cdc10-NCR region within mouse Notch 1.|
|Storage buffer||PBS, pH 7.4|
|Contains||0.09% sodium azide|
|Storage Conditions||4°C or -20°C if preferred|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||10 ul/10^6 cells|
|Immunohistochemistry (Frozen) (IHC (F))||1:50-1:200|
|Western Blot (WB)||1:200 - 1:1000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
|Immunoprecipitation (IP)||See 1 publications below|
Membrane permeabilization is required for flow cytometry applications. For FACS analysis, use 10ul of the suggested working dilution to label 1x10^6 cells in 100ul.
Notch1 is a single transmembrane receptor expressed in a range of cells including hematopoietic cells in mouse fetal liver, adult thymus and bone marrow. Notch1 plays a role in embryonic development and tissue homeostasis in the adult. Recent work has suggested a further role for this receptor in the regulation of lymphopoieisis and myelopoiesis. Following ligand binding, Notch1 is cleaved in the transmembrane domain, releasing the Notch intracellular domain (NICD) to translocate to the nucleus, where it plays a role in the modulation of gene expression.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Characterization of two distinct lymphoproliferative diseases caused by ectopic expression of the Notch ligand DLL4 on T cells.
MA1-81888 was used in immunoprecipitation to elucidate how Notch signaling contributes to T cell acute lymphoblastic leukemia using mice
|Xiong H,Maraver A,Latkowski JA,Henderson T,Schlessinger K,Ding Y,Shen J,Tadokoro CE,Lafaille JJ||PloS one (8:null)||2014|