Contents of the kit
Assay procedure
1. 1 bottle TRACER,32 ml, ready for use.
Contains less than 740 kBq of 125I labelled Label coated tubes in duplicate for each standard (S0-S5), control serum(C) and anti-SHBG and biotin labelled anti-SHBG in buffer containing proteins, 0.1% sodium Pipette 10 µl each of STANDARD(S0-
2. 6 vials STANDARD(S0-S5), ready for use.
0.5ml, human serum containing 0.1% NaN .
direct quantitative determination of SHBG in Pipette 300 µl of TRACER into each
range of 0-250 nmol/l using 10 µl serum 3. 1 vial CONTROL SERUM, lyophilized
samples. Each kit contains materials sufficient 0.5 ml human serum, containing 0.1% NaN3. with a plastic foil. If optional total counts construction of one standard curve and the specified in the quality certificate enclosed 4.. 2 boxes COATED TUBES, ready for use.
2X50 plastic tube, coated with streptavidin.
5. 1 bottle WASH BUFFER
adjust an adequate speed such that liquid Sex hormone-binding globulin (SHBG), also is constantly rotating or shaking in each Dilute with 700 ml distilled water before use. tube. Incubate tubes for 2 hours at room temperature. (Note: The efficient rotation protein (SBP), is a circulating glycoprotein is a critical factor to achieve good with a molecular weight of around 86000. It is performance. An uneven or incomplete thought to be synthesized in the liver, and in Materials, tools and equipment
shaking may result in a serious error. the circulation its biological function is the required
Never use a rack type with open hole.) transport of sex steroid hormones.It has a high tube and decant the supernatant from all disposable tips (10µl, 300µl, 2ml), shaker, tubes by the inversion of the rack. In the plastic foil, adsorbent tissue, gamma counter birth, increase to high levels during infancy, Recommended tools and equipment
then decrease during puberty. The highest repeating pipettes, dispenser with reservoir physiologic levels of SHBG are observed in Specimen collection and storage
Abnormal serum SHBG levels have been reported in number of conditions, including Calculation of results
Serum samples can be prepared according to common procedures used routinely in clinical Calculate the average CPM for each pair of (including polycystic ovary dicease). Serum laboratory practice. Samples can be stored at SHBG levels are inversely related to free, 2-8 °C if the assay is carried out within 48 plotting mean CPM of each standard level hours, otherwise aliquots should be prepared concentracions, a “free testosterone index” and stored deep frozen (-20°C). Do not store concentration, except for 0 standard (abscissa) based on the ratio of total testosterone to serum samples longer than 4 months. Do not use lipemic, hemolyzed or turbid specimens. Obtain sample concentration by interpolation Principle of method
thoroughly mixed before assaying. Repeated For computerised calculations and/or quality values, rather than cpm values are used. Preparation of reagents, storage
Specific binding values can be calculated for each standard and sample according to the antibodies react simultaneously with the opening. At this temperature reagent is stable antigen present in standards or samples which until expiry date. The actual expiry date is Assay Protocol, Pipetting Guide
leads to the formation of a capture antibody - given on the package label and in the quality Add 0.5 ml distilled water to the lyophilised control serum, and mix gently with shaking or immobilised on the reactive surface of test Ensure that complete dissolution is achieved, tubes. Reaction mixture is then discarded, test and allow the solution to equilibrate at room radioactivity is measured in a gamma counter. Add the wash buffer concentrate to 700 ml The concentration of antigen is directly distilled water. The diluted solution can be proportional to the radioactivity measured in stored at 2-8 °C until expiry date of the kit. test tubes. By constructing a calibration curve CAUTION!
plotting binding values against a series of Equilibrate all reagents and serum samples to Decant the fluid and blot on filter paper Decant the fluid and blot on filter paper Typical assay data
Chemical hazard
antimicrobial agent. Dispose of waste by flushing with copious amount of water to avoid build-up of explosive metallic azides in copper and lead plumbing. The total azide Recovery
increase expressed as per cent of expected increase upon spiking serum samples with known amount of SHBG. Values for 8 serum pooles spiked with SHBG at 3 levels were as Typical standard curve
Dilution test
4 samples were measured in a series of
dilution (2, 4, 8, 16-fold) with zero-standard. The following equation obtained for measured (Y) versus expected (X) concentration y =0.9301x - 0.5552, R=0.9962, n=16
Expected values
establish its own reference intervals. In a population (n=134) of adult female blood donors the mean (±SD) serum concentration Characterization of assay
In a population (n=139) of adult male blood donors the mean (±SD) serum concentration Calibration
Standards are calibrated against the WHO Results obtained should only be interpreted in Assay parame ters
the context of the overall clinical picture. None of in vitro diagnostic kits can be used as the one and only proof of any disease or Analytical sensitivity
The analytical sensitivity is 0.11nmol/l Additional information
It is defined as the concentration of SHBG Components from various lots or from kits of equivalent to the mean CPM of 20 replicates different manufacturers should not be mixed Functional sensitivity
The value of functional sensitivity is found to Radioactivity
It is defined as the value extrapolated to 20 % This product contains radioactive material. It of the inter-assay imprecision profile obtained is the responsibility of the user to ensure that from 10 independent runs on patient samples local regulations or code of practice related to the handling of radioactive materials are Hook effect
There is no high dose “hook effect” up to a Biohazard
Human blood products used in the kit have been obtained from healthy human donors. Specificity
Cross reactivity of the SHBG antiserum has been measured against human IgG (10 g/l) negative, for the presence of both Human In both cases cross reactivity non-detectable. Immunodeficiency Virus antibody (Anti-HIV- WEB site: http://www.izotop.hu
Technical e -mail: [email protected]
Precision and reproducibility
(HBsAg).Care should always be taken when Commercial e-mail: [email protected]
Samples with 20 replicates in 1 assay run, and handling human specimens to be tested with with duplicates in 12 runs were measured to diagnostic kits. Even if the subject has been precision, respectively. Values obtained are Immunodeficiency Virus (HIV-1), or other infectious agents are absent. Human blood Tel.: (+36) 1-392-2577, Fax: (+36) 1-395-9247 potentially infectious materials.

Source: http://www.izotop.hu/pdf/immuno/rk86ct_a.pdf

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