The Neural Cell Adhesion Molecule L1 (NCAM-L1) is an Immunoglobulin superfamily (IgSF) adhesion molecule critical for neural development. NCAM-L1 binds homophilically and heterophilically with adhesion molecules including Axonin-1, CD9, Neurocan, and Integrins. Mutations of the gene encoding NCAM-L1 are associated with a subset of X-linked mental retardation syndromes, schizophrenia, and other neurological disorders. Cancer cells can proteolytically cleave NCAM-L1, releasing the soluble ectodomain. Within the ECD, human NCAM-L1 shares 86% and 87% aa identity with mouse and rat NCAM-L1, respectively.
Suitable for use in Flow Cytometry. Other applications not tested.
Flow Cytometry: 10ul labels 1-2.5x10e5 cells. Validated using mouse splenocytes.
Optimal dilutions to be determined by the researcher.
Flow Cytometry Protocol:
1. Cells may be Fc-blocked with 1ug of mouse IgG/10e5 cells for 15 minutes at RT. Do not wash excess blocking IgG from this reaction.
2. After blocking, add 10ul of L0010-07 to 1-2.5x10e5 cells and incubate for 30 minutes at RT.
3. Remove unbound antibody by washing the cells twice in Flow Cytometry Staining Buffer (F4568-10). Note that whole blood requires a RBC lysis step at this point using Flow Cytometry Mouse Lyse Buffer.
4. Resuspend the cells in Flow Cytometry Staining Buffer for analysis by flow cytometry. As a control for this analysis, cells in a separate tube should be treated with PE-labeled rat IgG2a antibody.
Storage and Stability:
May be stored at 4 degrees C before opening. DO NOT FREEZE! Stable at 4 degrees C as an undiluted liquid. Stable for 12 months at 4 degrees C. Freezing R-Phycoerythrin (PE) conjugates will result in a substantial loss of activity. PE conjugates are sensitive to light.