Anti-Bone Sialoprotein II Antibody

Bone sialoprotein II is one of the major noncollagenous proteins in the extracellular matrix of bone. It is a phosphorylated glycoprotein with an approximate molecular weight of 70 kDa. The protein has been found in osteoblasts and osteocytes. Diseases concerning the bone turnover, pathological bone alterations as well as the rapid increase of osteoporosis make it necessary to establish new markers, Bone sialoprotein is discussed as a potential serum marker for monitoring bone remodelling.

Anti-Bone Sialoprotein II Antibody is an antibody against Bone Sialoprotein II for use in ELISA, IC, IH, IH(P), RIA & WB.

Western Blot
Immunohistochemistry (Paraffin) (see suggested protocol below)
Immunocytochemistry (1:100)
RIA
ELISA
Optimal working dilutions must be determined by the end user.

SUGGESTED STAINING PROCEDURE FOR RABBIT ANTI-BONE SIALOPROTEINAB1854
This protocol has been used successfully on 5 um, paraffin embedded sections from human fetal tibia, human fetal calvaria, human metastatic breast tumors to bone which were decalcified in 10% formic acid.

1. Dewaxing: Xylene 5 min. - Xylene 5 min. - Xylene 5 min.
2. Rehydration: 100% ethanol for 3 min. - 95% ethanol for 3 min. - 70% ethanol for 3 min.
3. Wash sections under running tap water for 5 minutes.
4. Wash sections in PBS.
5. Block endogenous peroxidase: 0.3% hydrogen peroxide in absolute methanol (2 mL of H2O2 30% in 198 mL methanol) for 30 minutes. (Cover the jar with foil, use stirring during incubation.)
6. Wash sections under running tap water for 10 minutes.
7. Mark tissue area with pap pen.
8. Cover the tissue area with 300 mL of blocking solution for 30 minutes at room temperature (blocking solution: 6 mL PBS, 0.3 gm BSA, 120 mL normal goat serum).
9. Shake off excess fluid (do not wash).
10. Incubate with primary antibody (AB1854 diluted 1:2,500 in PBS-T) at 2-8°C overnight in moist chamber.
11. Rinse with PBS-T, then wash 3 times in PBS-T each for 5 minutes at room temperature.
12. Incubate with biotinylated secondary antibody (for example Chemicon AP132B) for 30 minutes at room temperature.
13. Rinse with PBS-T, then wash in PBS-T 3 x 5 minutes at room temperature.
14. Incubate the sections with Vector ABC reagent for 30 minutes at room temperature. (ABC reagent: 2 drops of A and 2 drops of B, in 5 mL PBS, allow to stand 30 minutes at room temperature before use.)
15. Wash sections in PBS 3 x 5 minutes at room temperature.
16. Develop with DAB for 5-10 minutes.
17. Wash under running tap water.
18. Counter stain with Mayer′s HX for 45 seconds, wash in warm tap water.
19. Dehydrate: 70% ethanol - 95% ethanol - 100% ethanol, 1 min. each.
20. Wash in Xylene, mount, dry.

Human and rat Bone sialoprotein (BSP). No cross reactivity with human osteopontin or osteonectin by RIA or ELISA.

Other species have not been tested

Lyophilized. Resuspend in 100 μL aqua bidest. Contains no preservatives.

Maintain at 2-8°C (lyophilized) or -20°C (reconstituted) for up to 6 months in undiluted aliquots. Avoid repeated freeze/thaw cycles.

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