SPP1 Antibody

Rabbit anti-Osteopontin antibody, Bone sialoprotein 1, Secreted phosphoprotein 1, SPP-1, Urinary stone protein, Nephropontin, Uropontin, SPP1, BNSP, OPN

This whole rabbit serum was prepared by repeated immunizations with a synthetic peptide, from the human osteopontin protein, conjugated to KLH using maleimide.

Osteopontin is directed against human osteopontin. The antibody recognizes the full-length osteopontin protein (which runs at 66 kDa on westerns), as well as the C-terminal fragments of both thrombin and MMP-cleaved OPN. The 32 kDa MMP-cleaved C-fragment is recognized, but not the 40 kDa N-terminal fragment. Reactivity is reported to occur with osteopontin from swine, dog, mouse and rat.

Store vial at -20° C or below prior to opening. This vial contains a relatively low volume of reagent (25 μL) . To minimize loss of volume dilute 1:10 by adding 225 μL of the buffer stated above directly to the vial. Recap, mix thoroughly and briefly centrifuge to collect the volume at the bottom of the vial. Use this intermediate dilution when calculating final dilutions as recommended below. Store the vial at -20°C or below after dilution. Avoid cycles of freezing and thawing.

Anti-Osteopontin has been tested for western blotting, immunohistochemistry (formalin-fixed paraffin-embedded sections) and ELISA. The antibody exclusively recognizes C-terminal fragments of both thrombin and MMP-cleaved OPN. The antibody recognizes the full-length osteopontin protein (which runs at 66 kDa on westerns), and 32 kDa for the MMP-cleaved C-fragment, but the 40 kDa N-terminal fragment is not recognized. A 1:1000 dilution will detect strongly approximately 250 ng of OPN protein on a blot. A 1:100-1:300 dilution used for IHC on human breast tumor tissues. No pretreatment is required for IHC when formalin-fixed paraffin-embedded tissue is stained. Specific conditions for reactivity and signal detection should be optimized by the end user.