IGFBP4 ELISA Kit

Produktnummer ABIN3073435

Human IGFBP4 ELISA-Kit Colorimetric-Assay zur Quantifizierung von Human IGFBP4.

Kurzübersicht für IGFBP4 ELISA Kit (ABIN3073435)

Target: IGFBP4 (Insulin-Like Growth Factor Binding Protein 4 (IGFBP4))

Bindungsspezifität: total

Reaktivität: Human

Nachweismethode: Colorimetric

Methodentyp: Sandwich ELISA

Detektionsbereich: 9-255 ng/mL

Applikation: ELISA

Proben: Plasma, Serum

Produktdetails IGFBP4 ELISA Kit

Untere Nachweisgrenze: 9 ng/mL

Verwendungszweck: The Total IGFBP-4 enzyme linked immunosorbent assay (ELISA) kit provides materials for the quantitative measurement of IGFBP-4 in human serum, other biological fluids and cell culture supernatants. This kit is intended for laboratory research use only and is not for use in diagnostic or therapeutic procedures.

Analytische Methode: Quantitative

Sensitivität: 0.054 ng/mL

Bestandteile:

• IGFBP-4 Calibrators A
• Total IGFBP-4 Calibrators B-F (Lyophilized)
• Total IGFBP-4 Controls I & II
• IGFBP-4 Coated Microtitration strips
• IGFBP-4 Assay Buffer
• Total IGFBP-4 Biotin Conjugate Concentrate
• Total IGFBP-4 Streptavidin-Enzyme Conjugate-Ready-to-Use (RTU)
• TMB Chromogen Solution
• Stopping Solution
• Wash Concentrate A

Benötigtes Material:

1. Microplate absorbance 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. Optional related ELISA test kits for research purpose: a) picoPAPP-A ELISA b) Intact IGFBP-4 ELISA

Anwendungsinformationen

Probenmenge: 25 μL

Testdauer: 2.5 h

Plattentyp: Pre-coated

Aufbereitung der Reagenzien:

1. Total IGFBP-4 calibrators B-F and Total IGFBP-4 Controls I & II: Tap and reconstitute IGFBP-4 Calibrator B-F and IGFBP-4 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 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. Total IGFBP-4 Antibody-Biotin Conjugate Solution: The IGFBP-4 Antibody-Biotin Conjugate Concentrate should be diluted at a ratio of 1 part conjugate to 50 parts of IGFBP-4 Assay Buffer, according to the number of wells used. If an entire plate is to be used pipet exactly 220 μL of the Concentrate into 11 mL of the buffer.

Probennahme:

• Serum or plasma are 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 and mix thoroughly by gentle inversion before use. Calibrators, controls, and unknowns should be assayed in duplicate. NOTE: All samples that read higher than the highest calibrator should be diluted in the Calibrator A prior to the assay.

1. Reconstitute Total IGFBP-4 Calibrator B-F and Total IGFBP-4 Controls I & II each with 0.5 mL deionized water. Solubilize for 10 minutes, Mix well by gentle vortex.
2. Label the microtitration strips to be used.
3. Pipette 25 μL of the Calibrator, Controls and diluted samples to the appropriate wells.
4. Add 100 μL of the IGFBP-4 Assay Buffer 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 60 minutes at room temperature. (23+2oC)
6. With 30-40 minutes remaining of incubation time, prepare the Total IGFBP-4 Antibody-Biotin Conjugate Solution by diluting the Total IGFBP-4 Biotin Conjugate Concentrate (BCC-126) in the Assay buffer as described under the Preparation of the Reagents section of this insert.
7. Aspirate and wash each strip 5 times with Wash Solution (350 μL/per well) using an automatic microplate washer.
8. Add 100 μL of the IGFBP-4 Biotin Conjugate Solution (As specified in Preparation of Regents, step 4.0) 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 60 minutes at room temperature. (23+2oC)
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+2oC)
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 repeater
15. Incubate the wells, shaking at 600-800 rpm on an orbital microplate shaker, for 8-12 min at room temperature. NOTE: Visually monitor the color development to optimize the incubation time.
16. 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.

Ergebnisberechnung:

NOTE: The results in this package insert were calculated by plotting the linear optical density (OD) data on the y-axis and log Total IGFBP-4 concentration on X-axis using a non-linear regression curve-fit. Other data reduction methods may give slightly different results.

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

Testpräzision: Reproducibility of the Total IGFBP-4 assay was determined in a study using two serum pools for a total of 8 assays. At concentrations between 29.2 and 106.05ng/mL, the CV's ranged between 3.9 and 4.6%.

Beschränkungen: Nur für Forschungszwecke einsetzbar

Handhabung

Vorsichtsmaßnahmen: For Research Use Only. Not for use in diagnostic procedures. 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, which can be obtained at AnshLabs.com or by request.

Lagerung: 4 °C

Details zu IGFBP4

Target: IGFBP4 (Insulin-Like Growth Factor Binding Protein 4 (IGFBP4))

Andere Bezeichnung: IGFBP-4

Pathways: WNT Signalweg, Myometrial Relaxation and Contraction, Regulation of Carbohydrate Metabolic Process