Progesterone ELISA Kit

Produktnummer ABIN996932

Produktdetails Progesterone ELISA Kit

Target: Progesterone

Reaktivität: Hormone

Nachweismethode: Colorimetric

Methodentyp: Competition ELISA

Detektionsbereich: 0-50 ng/mL

Untere Nachweisgrenze: 0 ng/mL

Applikation: ELISA

Verwendungszweck: For the quantitative determination of Progesterone concentration in human serum or plasma.

Analytische Methode: Quantitative

Sensitivität: 0.06 ng/mL

Benötigtes Material: 1. Precision pipettes: 25 mL , 50 mL , 100 mL , 200 mL , and 1.0 mL .
2. Disposable pipette tips.
3. Distilled or deionized water.
4. Vortex mixer or equivalent.
5. Absorbent paper or paper towel.
6. Linear-linear graph paper.
7. Microtiter plate reader.

Anwendungsinformationen

Kommentare:

Quality Control:
1. Good laboratory practice requires that quality control specimens (controls) be run with each calibration curve - verify assay performance. - ensure proper performance, control material should be assayed repeatedly - establish mean values and acceptable ranges.
2. We recommend using Bio-Rad Lyphochek Immunoassay Control Sera as controls. The Diagnostic Automation, Inc. Progesterone EIA kit also is provided with internal controls, Levels 1 and
2.
3. Controls containing sodium azide cannot be used.
Limitations of procedure:
1. Reliable and reproducible results will be obtained when the assay procedure is carried out with a complete understanding of the package insert instructions and with adherence - good laboratory practice.
2. The wash procedure is critical. Insufficient washing will result in poor precision and falsely elevated absorbance readings.
3. Serum samples demonstrating gross lipemia, gross hemolysis, or turbidity should not be used with this test.
4. The results obtained from the use of this kit should be used only as an adjunct - other diagnostic procedures and information available - the physician. Cross-reactivity ( %) = x 100

Testdauer: 1.5 h

Plattentyp: Pre-coated

Aufbereitung der Reagenzien:

1. All reagents should be brought to room temperature (18-25 °C) before use.
   2. To prepare Working Progesterone-HRP Conjugate Reagent, add 0.1 mL of Progesterone-HRP Conjugate Concentrate (11x) to 1.0 mL of Progesterone-HRP Conjugate Diluent (1:10 dilution) and mix well. The amount of conjugate diluted depends on your assay size. Discard the excess after use.
   3. Samples with expected progesterone concentrations over 50 ng/mL may be quantitated by dilution with adequate diluent
   3. Use the mean absorbance values for each specimen to determine the corresponding concentration of Progesterone in ng/mL from the standard curve.
   4. Any values obtained for diluted samples must be further converted by applying the appropriate dilution factor in the calculations.

Aufbereitung der Proben:

1. Serum should be used in the test.
   2. No special pretreatment of sample is necessary.
   3. Serum samples may be stored at 2-8 °C for up to 24 hours, and should be frozen at -20 °C or lower for longer periods. Avoid grossly hemolytic (bright red), lipemic (milky), or turbid samples.
   4. Please note: Samples containing sodium azide should not be used in the assay.

Testdurchführung:

1. Secure the desired number of coated wells in the holder.
   2. Dispense
   2. mL of standards, specimens and controls into appropriate wells.
   3. Dispense 100 mL of Working Progesterone-HRP Conjugate Reagent into each well.
   4. Dispense 50 mL of rabbit anti-progesterone reagent to each well.
   5. Thoroughly mix for 30 seconds. It is very important to mix them completely.
   6. Incubate at room temperature (18-25 °C) for 90 minutes.
   7. Rinse and flick the microwells 5 times with distilled or deionized water. (Please do not use tap water.) 0.13 - 0.97 ng/mL 0.07 - 0.52 ng/mL 0.15 - 0.70 ng/mL 2.00 - 25.0 ng/mL 0.06 - 1.60 ng/mL
   8. Dispense 100 mL of TMB Reagent into each well. Gently mix for 10 seconds.
   9. Incubate at room temperature (18-25 °C) for
   2. minutes.
   10. Stop the reaction by adding 100 mL of Stop Solution to each well.
   11. Gently mix 30 seconds. It is important to make sure that all the blue color changes to yellow color completely.
   12. Read absorbance at 450 nm with a microtiter well reader within 15 minutes. 10.3-44.0 ng/mL 19.5-825 ng/mL 65.0-229 ng/mL

Ergebnisberechnung:

1. Calculate the mean absorbance value (A450) for each set of reference standards, controls and samples.
   
   2. Construct a standard curve by plotting the mean absorbance obtained for each reference standard against its concentration in ng/mL on a linear-linear graph paper, with absorbance values on the vertical or Y axis, and concentrations on the horizontal or X axis.
   a I. Accuracy A statistical study using 109 human serum samples demonstrated good correlation with a commercially available kit as shown below. Comparison between the Diagnostic Automation, Inc. Progesterone EIA and the DRG Progesterone kit provided the following data: N = 109 Correlation coefficient = 0.977 Slope = 0.867 Intercept = 0.727 DAI Mean = 5.3 ng/mL DRG Mean =
   3.9 ng/mL II.
   Each laboratory should establish its own normal range based on the patient population. Diagnostic Automation, Inc. Progesterone EIA was performed on rando mLy selected outpatient clinical laboratory samples. The following information is cited from reference #9. Males: adult Prepubertal (children) Females: follicular phase luteal phase post menopausal Pregnancy: Ftrmesier 2ndtrmesier 3rdTRMESIER
   Results of a typical standard run with optical density readings at 450 nm shown in the Y axis against Progesterone concentrations shown in the X axis. Note: This standard curve is for the purpose of illustration only, and should not be used to calculate unknowns. Each laboratory must provide its own data and standard curve in each experiment. 20 40 60 0 Progesterone Conc. (ng/mL)
   

Beschränkungen: Nur für Forschungszwecke einsetzbar

Handhabung

Haltbarkeit: 12-14 months

Details zu Progesterone

Target: Progesterone

Abstract: Progesterone Produkte

Substanzklasse: Hormone