The DSP (Dentin Sialoprotein) BioAssay™ ELISA Kit (Human) is a quantitative sandwich assay for the detection of DSP in human serum, plasma and other biological fluids. Cell lysates and tissue homogenates, though not tested, may potentially be used as samples.
Sensitivity:0.469ng/ml
Detection Range:0.781-50ng/ml
Specificity:This assay has high sensitivity and excellent specificity for detection of human DSP. No significant cross-reactivity or interference between DSP and analogues was observed.
Precision:Intra-Assay: CV<8%Inter-Assay: CV<10%
Test Principle:This ELISA kit employs the sandwich enzyme-linked immune-sorbent assay technology. Anti-DSP antibody was pre-coated onto 96-well plates. The biotin conjugated anti-DSP antibody was used as detection antibodies. The standards, test samples and biotin conjugated detection antibody were added to the wells subsequently, and wash with wash buffer. HRP-Streptavidin was added and unbound conjugates were washed away with wash buffer. TMB substrates were used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue colorproduct that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the DSP amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of DSP can be calculated.
Storage and Stability:Store unopened kit at 4ºC; store at -20ºC once opened. Store unopened standard at 4°C; once reconstituted, store at 4°C for up to 12 hours or at -20°C for up to 48 hours. Store other components at 4°C. Kit is stable for 6 months after receipt. For maximum recovery of product, centrifuge the original vials after thawing and prior to removing the cap.
Assay Summary:1. Wash plate 2 times before adding standards and samples to wells!2. Add 100ul standard or sample to each well and incubate for 90 minutes at 37°C. Aspirate and wash 2 times
3. Add 100ul Antibody (Biotin) working solution to each well and incubate for 60 minutes at 37°C. 4. Aspirate and wash 3 times
5. Add 100ul SABC working solution to each well. Incubate for 30 minutes at 37°C6. Aspirate and wash 5 times
7. Add 90ul TMB substrate. Incubate 10-20 minutes at 37°C8. Add 50ul Stop Solution. Read at 450nm immediately
