Chicken Thrombospondin- 2, THBS2 ELISA KIT

Thrombospondin was originally discovered as a secretory product of platelet alpha-granules. It is a multifunctional protein that contains binding sites for thrombin, fibrinogen, heparin, fibronectin, plasminogen, plasminogen activator, collagen, laminin, etc. It functions in many cell adhesion and migration events, including platelet aggregation. Thrombospondin-2 is a protein belongs to the 4 thrombospondin family. It is a disulfide-linked homotrimeric glycoprotein that mediates cell-to-cell and cell-to-matrix interactions. This protein has been shown to function as a potent inhibitor of tumor growth and angiogenesis. Studies of the mouse counterpart suggest that this protein may modulate the cell surface properties of mesenchymal cells and be involved in cell adhesion and migration.

【PRINCIPLE OF THE ASSAY】

This assay employs a two-site sandwich ELISA to quantitate THBS2 in Chicken serum, plasma. An antibody specific for THBS2 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any THBS2 present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for THBS2 is added to the wells. After washing, Streptavidin conjugated Horseradish Peroxidase (HRP) is added to the wells. Following a wash to remove any unbound avidin-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of THBS2 bound in the initial step. The color development is stopped and the intensity of the color is measured.
 

【LIMITATIONS OF THE PROCEDURE】

※ FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES.

※ The kit should not be used beyond the expiration date on the kit label.

※ Do not mix or substitute reagents with those from other lots or sources.

※ It is important that the Calibrator Diluent selected for the standard curve be

consistent with the samples being assayed.

※       If samples generate values higher than the highest standard, dilute the samples with the appropriate Calibrator Diluent and repeat the assay.

※       Any variation in standard diluent, operator, pipetting technique, washing

technique, incubation time or temperature, and kit age can cause variation in binding.

※       This assay is designed to eliminate interference by soluble receptors,

binding proteins, and other factors present in biological samples. Until all factors have been tested in the ELISA Kit, the possibility of interference cannot be excluded.

【DETECTION RANGE】

1.56 ng/mL - 100 ng/mL. The standard curve concentrations used for the ELISA’s were 100 ng/mL, 50 ng/mL, 25 ng/mL, 12.5 ng/mL, 6.25 ng/mL, 3.12 ng/mL, 1.56 ng/mL, 0 ng/mL.
 

【SENSITIVITY】

The limit of detection of Chicken THBS2 defined as the analyte concentration resulting in an absorbance significantly higher than that of the dilution medium (mean plus 2 standard deviations) was determined to be 1ng/mL (mean of 6 independent assays).
 

【SPECIFICITY】

This assay has high sensitivity and excellent specificity for detection of Chicken THBS2. No significant cross-reactivity or interference between Chicken THBS2 and analogues was observed.

Note:

Limited by current skills and knowledge, it is impossible for us to complete the cross-reactivity detection between Chicken THBS2 and all the analogues, therefore, cross reaction may still exist.
 

【PRECISION】

Intra-assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision. Inter-assay Precision (Precision between assays) Three samples of known concentration were tested in forty separate assays to assess inter-assay precision. CV (%) = SD/meanX100 Intra-Assay: CV<8%
Inter-Assay: CV<12%

【STABILITY】

The stability of ELISA kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.

The loss rate was determined by accelerated thermal degradation test. Keep the kit at 37°C for 4 and 7 days, and compare O.D.values of the kit kept at 37°C with that of at recommended temperature. (referring from Biological Products Standard, which was calculated by the Arrhenius equation. For ELISA kit, 4 days storage at 37°C can be considered as 6 months at 2-8°C, which means 7 days at 37°C equaling 12 months at 2-8°C).

Note:

To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Note:

1.This package insert must be read in its entirety before using this product.

2.This ELISA Kit is used for lab research only!