ADAM10 is a transmembrane multiple domain protein with ubiquitous expression. The metalloprotease activity of ADAM10 can cleave several molecules including Notch, pro-TNF-alpha, amyloid precursor protein, myelin basic protein, and type IV collagen.
Applications:
Suitable for use in Flow Cytometry. Other applications not tested.
Recommended Dilution:
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).
Optimal dilutions to be determined by the researcher.
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 before opening. DO NOT FREEZE! 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.