ADAM15 belongs to a family of transmembrane proteins that contain disintegrin and metalloprotease domains. Members of this family have been implicated in cell adhesion via integrin binding and in the proteolytic shedding of cell surface molecules.
Suitable for use in Flow Cytometry. Other applications not tested.
Flow Cytometry: Intracellular staining antibodies are designed for multiparameter flow
cytometric analysis of cells. To stain for surface proteins (e.g. CD3, CD4, CD8) in addition to the intracellular protein, we recommend that the investigator determine whether the fixation and permeabilization steps adversely affect the surface protein. If so, surface staining of cells prior to fixation and permeabilization is recommended.
For intracellular staining, cells must first be fixed and permeabilized. Use of 4% paraformaldehyde in PBS as a fixative is recommended. Other formulations or tissue fixatives may affect the staining properties of the monoclonal antibody. For permeabilization, 0.1% saponin in a balanced salt solution is effective in facilitating antibody entry into cells. Due to the reversible nature of cell membrane permeabilization, saponin must be included in all buffers used (i.e. both the staining and washing steps).
Additional Reagents Required:
Paraformaldehyde Fixative-Dissolve 4.0g of paraformaldehyde in 100ml of sterile PBS (10mM PBS, pH 7.4) by heating the solution at 56 degrees C for about 1 hour. All solids must be fully dissolved prior to use. Store buffer at 2-8 degrees C, protected from light, for no longer than 2 weeks.
SAP buffer-Prepare a sterile solution containing 0.1% (w/v) saponin, 0.05% (w/v) sodium azide.
Storage and Stability:
May be stored at 4 degrees C for short-term only. For long-term storage and to avoid repeated freezing and thawing, aliquot and store at -20 degrees C. Aliquots are stable for at least 12 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.