SHBG ELISA Kit

Catalog No. ABIN996939

Product Details SHBG ELISA Kit

Target: SHBG (Sex Hormone Binding Globulin (SHBG))

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 0-260 nM/mL

Minimum Detection Limit: 0 nM/mL

Application: ELISA

Purpose: SHBG ELISA is an enzyme immunoassay for the quantitative in vitro diagnostic measurement of SHBG in serum and heparin plasma.

Sample Type: Serum

Analytical Method: Quantitative

Specificity: 100%

Sensitivity: 0.77 nmol/L

Material not included: 1. A microtiter plate calibrated reader (450 ± 10 nm).
2. Calibrated variable precision micropipettes.
3. Absorbent paper.
4. Distilled or Deionized water.
5. Timer Semi logarithmic graph paper or software for data reduction.

Application Details

Comment:

Quality Control:
Good laboratory practice requires that controls be run with each calibration curve. A statistically significant number of controls should be assayed - establish mean values and acceptable ranges - assure proper performance. It is recommended - use control samples according - state and federal regulations. The use of control samples is advised - assure the day - day validity of results. Use controls at both normal and pathological levels. The controls and the corresponding results of the QC-Laboratory are stated in the QC certificate added - the kit. The values and ranges stated on the QC sheet always refer - the current kit lot and should be used for direct comparison of the results. It is also recommended - make use of national or international Quality Assessment programs in order - ensure the accuracy of the results. Employ appropriate statistical methods for analysing control values and trends. If the results of the assay do not fit - the established acceptable ranges of control materials patient results should be considered invalid. In this case, please check the following technical areas: Pipetting and timing devices, photometer, expiration dates of reagents, storage and incubation conditions, aspiration and washing methods. After checking the above mentioned items without finding any error contact DAI directly.
Limitations of procedure: Reliable and reproducible results will be obtained when the assay procedure is performed with a complete understanding of the package insert instruction and with adherence - good laboratory practice. Any improper handling of samples or modification of this test might influence the results. Interfering Substances Hemoglobin, bilirubin and triglycerides have no influence on the assay results. Drug Interference Until today no substances (drugs) are known - us, which have an influence - the measurement of SHBG in a sample. High-Dose Hook Effect Inter Assay Sample n Mean (nmol/L) CV ( %) 1 16 9.8 8.0 2 16 44.9 3.0 3 16 73.4 5.3 4 16 106.8 3.1 Recovery A known amount of SHBG was added - three patient sera and the quantities recovered were measured. The results are shown in the following table. No hook effect was observed in this test up - 40,000 nmol/L of SHBG.

Sample Volume: 10 μL

Assay Time: 1 h

Plate: Pre-coated

Reagent Preparation:

Bring all reagents and required number of strips to room temperature prior to use. Wash Solution
Add deionized water to the 40X concentrated Wash Solution.
Dilute 25 mL of concentrated Wash Solution with 975 mL deionized water to a final volume of 1000 mL.
The diluted Wash Solution is stable for 2 weeks at room temperature. Disposal of the kit The disposal of the kit must be made according to the national regulations. Special information for this product is given in the Material Safety Data Sheet. Damaged Test Kits In case of any severe damage to the test kit or components, DAI has to be informed in writing, at the latest, one week after receiving the kit. Severely damaged single components should not be used for a test run. They have to be stored until a final solution has been found. After this, they should be disposed according to the official regulations.

Sample Preparation:

Serum or heparin plasma can be used in this assay.
EDTA-plasma may give slightly lower results.
Do not use haemolytic, icteric or lipaemic specimens.
Please note: Samples containing sodium azide should not be used in the assay. See external Label 96 Tests Test Sex-hormone-binding globulin Method Enzyme Linked Immunosorbent Assay Principle Sandwich Complex Detection Range 0.77-260 nmol/L Sample 10 μL serum / plasma Total Time -60 min. Shelf Life 12-14 Months from the manufacturing date Specificity 100 %

Assay Procedure:

1. General Remarks
   1. All reagents and specimens must be allowed to come to room temperature before use. All reagents must be mixed without foaming.
   2. Once the test has been started, all steps should be completed without interruption.
   3. Use new disposal plastic pipette tips for each standard, control or sample in order to avoid cross contamination.
   4. Absorbance is a function of the incubation time and temperature. Before starting the assay, it is recommended that all reagents are ready, caps removed, all needed wells secured in holder, etc. This will ensure equal elapsed time for each pipetting step without interruption.
   5. As a general rule the enzymatic reaction is linearly proportional to time and temperature. Pipetting of samples should not exceed 10 minutes to avoid assay drift. If more than one plate is used in the same run, it is recommended to include a standard curve on each plate. Each run must include a standard curve.
   
   Test Procedure: Each run must include a standard curve. Add 100 μL of Substrate Solution to each well. Incubate for 12 minutes at room temperature for 8 minutes at room temperature (26 °C and more). Stop the enzymatic reaction by adding 100 μL of Stop Solution to each well. Determine the absorbance (OD) of each well at 450 + 10 nm with a microtiter plate reader. It is recommended that the wells be read within 10 minutes after adding the Stop Solution.

Calculation of Results:

1. Calculate the average absorbance values for each of standards, controls and samples.
   
   2. Using semi-logarithmic graph paper, construct a standard curve by plotting the mean absorbance obtained from each standard against its concentration with absorbance value on the vertical (Y) axis and concentration on the horizontal (X) axis.
   
   3. Using the mean absorbance value for each sample determine the corresponding concentration from the standard curve.
   4. Automated method: The results in the IFU have been calculated automatically using a 4 PL (4 Parameter Logistics) curve fit. 4 Parameter logistics is the preferred method. Other data reduction functions may give slightly different results.
   5. The concentration of the samples can be read directly from this standard curve. Samples with concentrations higher than that of the highest standard have to be further diluted or reported as > 260 nmol/L. For the calculation of the concentrations this dilution factor has to be taken into account. Example of Typical Standard curve The following data is for demonstration only and cannot be used in place of data generation s at the time of assay. Standard Optical Units (450 nm) Standard 0 (0 nmol/L) 0.01 Standard 1 (4 nmol/L) 0.08 Standard 2 (16 nmol/L) 0.30 Standard 3 (65 nmol/L)
   1.07 Standard 4 (260 nmol/L)
   2.04 EXPECTED NORMAL VALUES It is strongly recommended that each laboratory should determine its own normal and abnormal values. In a study conducted with apparently normal healthy adults, using the DAI SHBG ELISA the following values are observed:
   1. Secure the desired number of Microtiter wells in the frame holder.
   
   2. Dilute each Standard, Control and sample 1:20 with Assay Buffer (1 part Standards/Control/sample + 19 parts Assay Buffer) Example: 10 μL Standard + 190 μL Assay Buffer
   
   3. Dispense 100 μL Assay Buffer into each well.
   4. Dispense 25 μL of each diluted Standard, Control and sample with new disposable tips into appropriate wells. Thoroughly mix for 5 seconds. It is important to have a complete mixing in this step.
   5. Incubate for 30 minutes at room temperature.
   6. Briskly shake out the contents of the wells. Rinse the wells 3 times with diluted Wash Solution (300-400 μL per well). Strike the wells sharply on absorbent paper to remove residual droplets. Important note: The sensitivity and precision of this assay is markedly influenced by the correct performance of the washing procedure!
   7. Dispense 100 μL Enzyme Conjugate into each well.
   8. Incubate for 15 minutes at room temperature.
   9. Briskly shake out the contents of the wells. Rinse the wells 3 times with diluted Wash Solution (300-400 μL per well). Strike the wells sharply on absorbent paper to remove residual droplets. Population N SHBG nmol/L Mean range Males 102 43 15-100 Females 44 62 15-120 The results alone should not be the only reason for any therapeutic consequences. The results should be correlated to other clinical observations and diagnostic tests.
   Assay Dynamic Range The range of the assay is between 0.77-260 nmol/L. Specificity of Antibodies (Cross Reactivity) Specificity of the SHBG ELISA was studied by measuring apparent SHBG response caused by high levels of TBG (Thyroxine Binding Globulin) and CBG (Cortisol Binding Globulin). No cross-reactions were found when testing up to 500 mg/L of TBG and 500 mg/L of CBG.
   

Restrictions: For Research Use only

Handling

Storage: 4 °C

Expiry Date: 12-14 months

Target Details for SHBG

Target: SHBG (Sex Hormone Binding Globulin (SHBG))

Alternative Name: SHBG (SHBG Products)

Synonyms: ABP ELISA Kit, SBP ELISA Kit, TEBG ELISA Kit, Abpa ELISA Kit, shbgl ELISA Kit, wu:fb66h07 ELISA Kit, wu:fd55b08 ELISA Kit, zgc:123063 ELISA Kit, shbg ELISA Kit, sex hormone binding globulin ELISA Kit, sex hormone-binding globulin ELISA Kit, sex hormone binding globulin L homeolog ELISA Kit, SHBG ELISA Kit, Shbg ELISA Kit, shbg ELISA Kit, shbg.L ELISA Kit

Pathways: Hormone Transport