Activin B ELISA Kit

Produktnummer ABIN5680716

Produktdetails Activin B ELISA Kit

Target: Activin B (ACVB)

Reaktivität: Human

Nachweismethode: Colorimetric

Methodentyp: Sandwich ELISA

Detektionsbereich: 12.7 pg/mL - 1400 pg/mL

Untere Nachweisgrenze: 12.7 pg/mL

Applikation: ELISA

Verwendungszweck: The Activin B enzyme linked immunuosorbent assay (ELISA) kit provides materials for the quantitative measurement of Activin B in human serum, lithium heparin plasma and other biological fluids.

Proben: Plasma, Serum

Analytische Methode: Quantitative

Sensitivität: 4.35 pg/mL

Bestandteile:

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

Benötigtes Material:

1. Microplate reader capable of absorbance measurement at 450 nm, 405nm 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.

Anwendungsinformationen

Probenmenge: 25 μL

Testdauer: 3.5 h

Plattentyp: Pre-coated

Aufbereitung der Reagenzien:

1. Activin B Calibrators B-F and Activin B Controls I & II: Tap and reconstitute Activin B Calibrator B-F and Activin B Controls I & II each with 0.5 mL deionized water. Solubilize, 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. Activin B Antibody-Biotin Conjugate Solution: The Activin B Antibody-Biotin Conjugate Concentrate should be diluted at a ratio of 1 part conjugate to 50 parts of Activin B 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.

Probennahme:

• 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.

Testdurchführung:

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: 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 Activin B Calibrator B-F and Activin B Controls I & II each with 0.5 mL deionized water. Solubilize for 10 minutes, Mix well.
2. Label the microtitration strips to be used.
3. Pipette 25 μL of the Calibrator, Controls and Unknowns to the appropriate wells.
4. Add 50 μL of the Activin A Assay Buffer A to each well using a repeater pipette.
5. Incubate the plate, shaking at a fast speed (600-800 rpm) on an orbital microplate shaker, for 2 hour at room temperature (23 ± 2 °C).
6. During the last 20-30 minutes of incubation, prepare the Activin B Antibody-Biotin Conjugate Solution by diluting the Activin B Biotin Conjugate Concentrate in Activin B Conjugate Diluent as described under the Preparation of the Reagents section of this package insert.
7. Aspirate and wash each strip 5 times with Washing Solution (350 μL/per well) using an automatic microplate washer.
8. Add 100 μL of the Antibody-Biotin Conjugate Solution to each well using a repeater pipette.
9. 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).
10. Aspirate and wash each strip 5 times with the Wash Solution (350 μL/per well) using an automatic microplate washer.
11. Add 100 μL of the Streptavidin-Enzyme Conjugate-RTU to each well using a repeater pipette.
12. 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).
13. Aspirate and wash each strip 5 times with the Wash Solution (350 μL/per well) using an automatic microplate washer.
14. Add 100 μL of the TMB chromogen solution to each well using a precision pipette. Avoid exposure to direct sunlight.
15. Incubate the wells, shaking at 600-800 rpm on an orbital microplate shaker, for 8-12 min at room temperature (23 ± 2 °C). NOTE: Visually monitor the color development to optimize the incubation time.
16. 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"".

Ergebnisberechnung:

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. 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 Activin B concentrations in pg/mL along the x-axis, using a cubic regression curve-fit.
4. Determine the Activin B concentrations of the Controls and unknowns from the calibration curve by matching their mean OD readings with the corresponding Activin B 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.

Beschränkungen: Nur für Forschungszwecke einsetzbar

Handhabung

Vorsichtsmaßnahmen: 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 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 may 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.

Lagerung: 4 °C

Details zu Activin B

Target: Activin B (ACVB)

Andere Bezeichnung: Activin B (ACVB Produkte)

Synonyme: inhibin ELISA Kit, inhibin beta B L homeolog ELISA Kit, inhibin beta B ELISA Kit, inhbb.L ELISA Kit, inhbb ELISA Kit