Inhibin alpha ELISA Kit

Catalog No. ABIN5680707

Product Details Inhibin alpha ELISA Kit

Target: Inhibin alpha (INHA) (Inhibin, alpha (INHA))

Reactivity: Mouse, Rat, Dog, Horse

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 6 pg/mL - 668 pg/mL

Minimum Detection Limit: 6 pg/mL

Application: ELISA

Purpose: The animal Inhibin A enzyme linked immunuosorbent assay (ELISA) kit provides materials for the quantitative measurement of Inhibin A in serum and other biological fluids.

Sample Type: Plasma, Serum

Analytical Method: Quantitative

Sensitivity: 2.3 pg/mL

Components:

• Inhibin A Calibrators A
• Inhibin A Calibrators B thru F (Lyophilized)
• Inhibin A Controls I & II (Lyophilized)
• Inhibin A Coated Microtitration Strips
• Inhibin A Assay Buffer A
• Inhibin A Assay Buffer B
• Inhibin A Biotin Conjugate Concentrate
• Inhibin A Biotin Conjugate Diluent
• Inhibin A 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.
7. Disposable 12 x 75 mm culture tubes.
8. Tight fitting 12 x 75 mm tube racks.

Application Details

Sample Volume: 15 μL

Assay Time: 4 h

Plate: Pre-coated

Reagent Preparation:

1. Inhibin A Calibrators B-F and Inhibin A Controls I & II: Tap and reconstitute Inhibin A Calibrators B-F and Inhibin A Controls I & II each with 1 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.
4. Inhibin A Assay Buffers Premix Solution: The Inhibin A Assay Buffer A (ASB-123A) and Inhibin A Assay Buffer B (ASB-123B) should be mixed by gentle inversion in equal volumes (1:1 ratio) according to the number of wells used. If an entire plate is to be used mix exactly 3 mL of the ASB-123 B in to 3 mL of the ASB-123 A. The pre-mixture solution is stable for use up to 4 hours. Discard the pre-mix solution after 4 hours.
5. Inhibin A Antibody-Biotin Conjugate Solution: The Inhibin A Antibody-Biotin Conjugate Concentrate should be diluted at a ratio of 1 part conjugate to 50 parts of Inhibin A Conjugate Diluent, according to the number of wells used. If an entire plate is to be used pipet exactly 220 μL of the Concentrate in to 11 mL of the diluent.

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.
• 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. 14

Assay Procedure:

Allow all specimens and reagents to reach room temperature (23 ± 2 °C) and mix thoroughly by gentle inversion before use. Calibrators, controls, and unknowns should be assayed in duplicate. NOTE: i. All intact mouse and rat serum samples should be diluted 1 part in 3 parts (1:4) in Calibrator A/Sample diluent prior to assay. ii. 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. Label the microtitration strips to be used.
2. Pipette 50 μL of the Calibrator, Controls and Unknowns to the appropriate wells.
3. Add 50 μL of Inhibin A Assay Buffer premix (ASB-123A and ASB-123B in 1:1 ratio as described under the Preparation of the Reagents section of this package insert) to each well using a repeater pipette.
4. Incubate the plate, shaking at a fast speed (600-800 rpm) on an orbital microplate shaker, for 2.5 hour at room temperature (23 ± 2 °C).
5. During the last 20-30 minutes of incubation, prepare the Inhibin A Antibody-Biotin Conjugate Solution by diluting the Inhibin A Biotin Conjugate Concentrate in Inhibin A Conjugate Diluent as described under the Preparation of the Reagents section of this package insert.
6. Aspirate and wash each strip 5 times with Washing Solution (350 μL/per well) using an automatic microplate washer.
7. Add 100 μL of the Antibody-Biotin Conjugate Solution to each well using a repeater pipette.
8. Incubate the plate, shaking at a fast speed (600-800 rpm) on an orbital microplate shaker, for 1 hour at room temperature (23 ± 2 °C).
9. Aspirate and wash each strip 5 times with the Wash Solution (350 μL/per well) using an automatic microplate washer.
10. Add 100 μL of the Streptavidin-Enzyme Conjugate-RTU to each well using a repeater pipette.
11. Incubate the plate, shaking at a fast speed (600-800 rpm) on an orbital microplate shaker, for 30 minutes at room temperature (23 ± 2 °C).
12. Aspirate and wash each strip 5 times with the Wash Solution (350 μL/per well) using an automatic microplate washer.
13. Add 100 μL of the TMB chromogen solution to each well using a repeater pipette. Avoid exposure to direct sunlight.
14. Incubate the wells, shaking at 600-800 rpm on an orbital microplate shaker, for 12-15 min at room temperature (23 ± 2 °C). NOTE: Visually monitor the color development to optimize the incubation time.
15. Add 100 μL of the stopping solution to each well using a repeater pipette. Read the absorbance of the solution in the wells within 20 minutes, using a microplate reader set to 450 nm. NOTE: Zero calibrator should be programmed as ""Blank"" while reading the optical density. If instrument has a wavelength correction, set the instrument to dual wavelength measurement at 450 nm with background wavelength correction at 630 nm.

Calculation of Results:

NOTE: The results in this package insert were calculated by plotting the log optical density (OD) data on the y-axis and log Inhibin A concentration on X-axis using a cubic regression curve-fit. Alternatively, log vs. log quadratic regression curve-fit can be used. Other data reduction methods may give slightly different results.

1. Optimum results can be obtained at incubation temperature of 23 ± 2 °C.
2. Calculate the mean OD for each calibrator, Control, or Unknown.
3. Plot the log of the mean OD readings for each of the Calibrators along the y-axis versus log of the Inhibin A concentrations in pg/mL along the x-axis, using a linear regression curve-fit.
4. Determine the Inhibin A concentrations of the Controls and unknowns from the calibration curve by matching their mean OD readings with the corresponding Inhibin A concentrations.
5. Any sample reading higher than the highest Calibrator should be appropriately diluted with the 0 pg/mL (CAL A) and re-assayed.
6. Any sample reading lower than the analytical sensitivity should be reported as such.
7. Multiply the value by a dilution factor, if required.

Restrictions: For Research Use only

Handling

Precaution of Use: 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. d) If external package is damaged, inspect the components inside for any other damage. Do not use if the components are damaged. WARNING: Potential Biohazardous Material Samples and blood-derived products may be routinely processed with minimum risk using the procedure described. However, handle these products as potentially infectious according to universal precautions and good clinical laboratory practices, regardless of their origin, treatment or prior certification.15 Use an appropriate disinfectant for decontamination. Store and dispose of these materials and their containers in accordance with local regulations and guidelines. WARNING: Potential Chemical Hazard Some reagents in this kit contain Pro-Clean 400 and Sodium azide16 as a preservative. Pro-Clean 400 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 alpha

Target: Inhibin alpha (INHA) (Inhibin, alpha (INHA))

Alternative Name: Inhibin A (INHA Products)

Synonyms: si:dkey-91f15.2 ELISA Kit, INHA ELISA Kit, MGC108500 ELISA Kit, AW555078 ELISA Kit, inhibin, alpha ELISA Kit, inhibin alpha subunit ELISA Kit, inhibin alpha ELISA Kit, inhibin alpha L homeolog ELISA Kit, inha ELISA Kit, INHA ELISA Kit, inha.L ELISA Kit, Inha ELISA Kit

Pathways: Peptide Hormone Metabolism, Hormone Activity, Negative Regulation of Hormone Secretion