Bone Sialoprotein (BSP) ELISA Assay

$1,720.00

The Bone Sialoprotein (BSP) ELISA Assay Kit is an enzyme immunoassay intended for the quantitative determination of underglycosylated bone sialoprotein (BSP) in serum and EDTA plasma. For research use only. Not for use in diagnostic procedures.

SKU: KR4222 Categories: , ,

Bone Sialoprotein BSP ELISA Assay

Bone Sialoprotein BSP ELISA Assay manufactured in Germany by Immundiagnostik

Size: 1×96 wells
Standard Range: 3.1-50 ng/ml
Incubation Time: 3 h, 1 h, 10-20 min
Sample Type: Serum, EDTA Plasma
Sample Size: 50 µL
For Research Use Only
Controls Included


Scientific Background

Bone sialoprotein (BSP) is a phosphorylated glycoprotein that comprises 12 % of the total non-collagenous proteins in bone and is thought to be involved in bone mineralisation and remodelling. The polypeptide chain of BSP has a molecular weight of 35 kDa. Native BSP has an apparent molecular weight of 70–80 kDa due to complex post translational modifications involving glycosylation. BSP is synthesized mainly by skeletal-associated cell types, including fibroblasts, hypertrophic chondrocytes, osteoblasts, osteocytes, osteoclasts, as well as trophoblasts. BSP is overexpressed and underglycosylated in human primary breast and prostate cancers that metastasize to the bone, whereby BSP expression correlates with the development of microcalcifications. Underglycosylated BSP is also expressed by other tumours, such as lung cancer, thyroid cancer, multiple myeloma and neuroblastoma, that predominantly metastasise to bones.


Assay Principle


This ELISA is designed for the quantitative determination of underglycosylated bone sialoprotein in serum and EDTA plasma. The test principle is based on interaction of the antigen in the samples or standards and with the antibody coated on the wells of the microtiter plate. A peroxidase-conjugated secondary antibody is used for detection and quantification, and tetramethylbenzidine (TMB) as a peroxidase substrate. The enzymatic reaction is terminated by an acidic stop solution. A dose response curve of the absorbance unit (optical density, OD at 450 nm) vs. concentration is generated, using the values obtained from the standard. Bone sialoprotein, present in the patient samples, is determined directly from this curve.

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Additional Information

Storage and Preparation of Reagents
• To run the assay more than once, ensure that reagents are stored at the conditions stated on the label. Prepare only the appropriate amount necessary for each run. The kit can be used up to 4 times within the expiry date stated on the label.
• Reagents with a volume less than 100µl should be centrifuged before use to avoid loss of volume.
• Preparation of the wash buffer: The wash buffer concentrate (WASHBUF) has to be diluted with ultra pure water 1:50 before use (20ml WASHBUF + 980ml ultra pure water), mix well. Crystals could occur due to high salt concentration in the concentrate. Before dilution, the crystals have to be redissolved at room temperature or in a water bath at 37°C. The WASHBUF is stable at 2–8°C until the expiry date stated on the label. Wash buffer (1:50 diluted WASHBUF) can be stored in a closed flask at 2–8°C for 1 month.
• The lyophilised standards (STD) and controls (CTRL) are stable at 2–8°C until the expiry date stated on the label. Before use, the STD and CTRL have to be reconstituted with 500µl of ultra pure water. Allow the vial content to dissolve for 10 minutes and mix thoroughly to ensure complete reconstitution. Standards and controls (reconstituted STD and CTRL) can be stored at -20°C for 3 weeks. Avoid repeated thawing and freezing.
• Preparation of the conjugate: Before use, the conjugate concentrate (CONJ) has to be diluted 1:101 in wash buffer (100µl CONJ + 10ml wash buffer). The CONJ is stable at 2–8°C until the expiry date stated on the label. Conjugate (1:101 diluted CONJ) is not stable and cannot be stored.
• All other test reagents are ready-to-use. Test reagents are stable until the expiry date (see label) when stored at 2–8°C.


Manual

Product Manual