Inhibin ELISA Kit

Catalog No. ABIN3073444

Product Details Inhibin ELISA Kit

Target: Inhibin (INH)

Binding Specificity: total

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 9-1000 pg/mL

Minimum Detection Limit: 9 pg/mL

Application: ELISA

Purpose: The Total Inhibin enzyme linked immunosorbent assay (ELISA) kit provides materials for the quantitative measurement of Total inhibin (Inhibin A, Inhibin B and Inhibin alpha subunit) in human serum, plasma and other biological fluids. This kit is intended for laboratory Research Use Only and is not for use in diagnostic or therapeutic procedures.

Sample Type: Plasma, Serum

Analytical Method: Quantitative

Sensitivity: 1.5 pg/mL

Components:

• Total Inhibin Calibrators A (Lyophilized)
• Total Inhibin Calibrators B thru F (Lyophilized)
• Total Inhibin Controls I & II (Lyophilized)
• Total Inhibin antibody Coated Microtitration Strips
• Inhibin B Assay Buffer A
• Inhibin B Assay Buffer B
• Total Inhibin Biotin Conjugate Concentrate
• Inhibin B Conjugate Diluent
• Total Inhibin Streptavidin-Enzyme Conjugate-Ready-to-Use (RTU)
• TMB Chromogen Solution
• Stopping Solution
• Wash Concentrate A

Material not included:

1. Microplate reader capable of absorbance measurement at 450 nm, 405 nm and 630 nm.
2. Microplate orbital shaker.
3. Microplate washer.
4. Semi-automated/manual precision pipette to deliver 10-250 μL.
5. Vortex mixer.
6. Deionized water.

Application Details

Sample Volume: 50 μL

Assay Time: 3.5 h

Plate: Pre-coated

Reagent Preparation:

1. IGF-II Calibrators A-F and IGF-II Controls I & II: Tap and reconstitute IGF-II Calibrators A-F and IGF-II Controls I & II each with 1.0 mL deionized water. Solubilize for 10 minutes, mix well and use after reconstitution.
2. Wash Solution: Dilute wash concentrate 25-fold with deionized water. The wash solution is stable for one month at room temperature (23± 2 °C) when stored in a tightly sealed bottle.
3. Microtitration Wells: Select the number of coated wells required for the assay. The remaining unused wells should be placed in the resealable pouch with a desiccant. The pouch must be resealed to protect from moisture.

Sample Collection:

• Serum is the recommended sample type.
• Sample handling, processing, and storage requirements depend on the brand of blood collection tube that you use. Please reference the manufacturer's instructions for guidance. Each laboratory should determine the acceptability of its own blood collection tubes and serum separation products.
• Samples may be stored at 4 °C if assayed within 24 hours, otherwise samples must be stored at -20 °C or -80 °C to avoid loss of bioactivity and contamination.
• Avoid assaying lipemic, hemolyzed or icteric samples.
• Avoid repeated freezing and thawing of samples. Thaw samples no more than 3 times.
• For shipping, place specimens in leak proof containers in biohazard specimen bags with appropriate specimen identification and test requisition information in the outside pocket of the biohazard specimen bag. Follow DOT and IATA requirements when shipping specimens

Assay Procedure:

Allow all specimens and reagents to reach room temperature and mix thoroughly by gentle inversion before use. Calibrators, controls, and unknowns should be assayed in duplicate. NOTE: All serum samples reading higher than the highest calibrator should be mixed and diluted in the 0 pg/mL reconstituted Calibrator A prior to assay.

1. Reconstitute Total Inhibin Calibrators A-F and Total Inhibin Controls I & II each with 1.0 mL deionized water. Solubilize for 10 minutes, Mix well.
2. Label the microtitration strips to be used.
3. Pipette 50 μL of the Calibrator, Controls and Unknowns to the appropriate wells.
4. Add 50 μL of Inhibin B Assay Buffer A to each well using a repeater pipette.
5. Add 50 μL of the Inhibin B Assay Buffer B to each well using a repeater pipette.
6. Incubate the plate, shaking at a fast speed (600-800 rpm) on an orbital microplate shaker, for 2 hours at room temperature.
7. During the last 20-30 minutes of incubation, prepare the Total Inhibin Antibody-Biotin Conjugate Solution by diluting the Total Inhibin Biotin Conjugate Concentrate in Inhibin B Conjugate Diluent as described under the Preparation of the Reagents section of this package insert.
8. Aspirate and wash each strip 5 times with Washing Solution (350 μL/per well) using an automatic microplate washer.
9. Add 100 μL of the Antibody-Biotin Conjugate Solution to each well using a repeater pipette.
10. Incubate the plate, shaking at a fast speed (600-800 rpm) on an orbital microplate shaker, for 1 hour at room temperature.
11. Aspirate and wash each strip 5 times with the Wash Solution (350 μL/per well) using an automatic microplate washer.
12. Add 100 μL of the Streptavidin-Enzyme Conjugate-RTU to each well using a repeater pipette.
13. Incubate the plate, shaking at a fast speed (600-800 rpm) on an orbital microplate shaker, for 30 minutes at room temperature.
14. Aspirate and wash each strip 5 times with the Wash Solution (350 μL/per well) using an automatic microplate washer.
15. Add 100 μL of the TMB chromogen solution to each well using a precision pipette. Avoid exposure to direct sunlight.
16. Incubate the wells, shaking at 600-800 rpm on an orbital microplate shaker, for 10-12 min at room temperature. NOTE: Visually monitor the color development to optimize the incubation time.
17. Add 100 μL of the stopping solution to each well using a precision pipette. Read the absorbance of the solution in the wells within 20 minutes, using a microplate reader set to 450 nm. NOTE: While reading the absorbance of the microtitration well, it is necessary to program the zero calibrator as a "Blank".

Calculation of Results:

NOTE: The results in this package insert were calculated by plotting the data on a log vs. log scale using a cubic regression curve-fit. Other data reduction methods may give slightly different results.

1. Calculate the mean OD for each calibrator, Control, or Unknown.
2. Plot the log of the mean OD readings for each of the Calibrators along the y- axis versus log of the Total Inhibin concentrations in pg/mL along the x-axis, using a cubic regression curve-fit.
3. Determine the Total Inhibin concentrations of the Controls and unknons from the calibration curve by matching their mean OD readings with the corresponding Total Inhibin concentrations.
4. Any sample reading higher than the highest Calibrator should be appropriately diluted with the 0 pg/mL (CAL A) and re-assayed.
5. Any sample reading lower than the analytical sensitivity should be reported as such.
6. Multiply the value by a dilution factor, if required.

Assay Precision: Reproducibility of the Total Inhibin assay was determined in a study using three serum pools and two kit controls for a total of nine assays. At concentrations between 20.5 and 174.5ng/mL, the CV's ranged between 2.6 and 10.01%.

Restrictions: For Research Use only

Handling

Precaution of Use: For research use only. The following precautions should be observed: a) Follow good laboratory practice. b) Use personal protective equipment. Wear lab coats and disposable gloves when handling immunoassay materials. c) Handle and dispose of all reagents and material in compliance with applicable regulations. WARNING: Potential Biohazardous Material This reagent may contain some human source material (e.g. serum) or materials used in conjunction with human source materials. Handle all reagents and patient samples at a Biosafety Level 2, as recommended for any potentially infectious human material in the Centers for Disease Control/National Institutes of Health manual "Biosafety in Microbiological and Biomedical Laboratories," 5th Edition, 2007. WARNING: Potential Chemical Hazard Some reagents in this kit contain Pro-Clean 400 and Sodium Azide as a preservative. Pro-Clean 400 and Sodium Azide in concentrated amounts are irritants to skin and mucous membranes. For further information regarding hazardous substances in the kit, please refer to the MSDS.

Storage: 4 °C

Target Details for Inhibin

Target: Inhibin (INH)

Alternative Name: Inhibin (INH Products)

Synonyms: inhibin ELISA Kit, inhibin beta B L homeolog ELISA Kit, inhibin beta B ELISA Kit, inhibin ELISA Kit, inhbb.L ELISA Kit, inhbb ELISA Kit, inh ELISA Kit