Mouse F4/80 glycoprotein contains seven-transmembrane (TM7) regions, which anchor the protein in the cell membrane, and thereby shows similarity in this region to G-protein-coupled receptors. The F4/80 molecule shares overall structural homology to other members of the epidermal growth factor (EGF)-TM7 family. The antigen is detected on tissue fixed macrophages in all organs tested so far (spleen, lymph nodes, thymus, liver, skin). It is also present on Langerhans cells in the skin and Kupffer cells in the liver. It is absent on granulocytes, lymphocytes and trombocytes. The expression of F4/80 increases upon maturation of macrophage precursors in bone marrow and blood as well as in ontogeny.
Applications:
Suitable for use in Flow Cytometry, Western Blot and Immunohistochemistry. Other applications not tested.
Recommended Dilution:
Flow Cytometry: 1:50, Fix with 1% paraformaldehyde (3).
Western Blot: 1:50, Mouse bone-marrow derived macrophages; non-reduced. Detects a band at ~125kD (3). Reduction with 2-mercaptoethanol destroys the antigen.
Immunohistochemistry (frozen): 1:50, Tissue embedded in OCT Tissue Tec; fixed with acetone for 10 min at RT. Incubation with 0.02M sodium azide in PBS, 0.1% H2O2 for 10min at RT to destroy endogenous peroxidase. Use spleen as positive control
Immunohistochemistry (paraffin): 1:50, Fixation in 10% neutral buffered formalin for 24 hours; blocking with non-immunized goat serum; microwaved for 6min in citrate buffer. Use splenic macrophages as positive control (5).
Optimal dilutions to be determined by the researcher.
Positive Control: Mouse macrophages
Negative Control: Mouse fibroblasts or granulocytes
Storage and Stability:
May be stored at 4 degrees C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20 degrees C. Aliquots are stable for 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. FITC conjugates are sensitive to light.