XDH ELISA Kit

Catalog No. ABIN6222583

This Colorimetric ELISA kit is designed for the quantitative measurement of Human XDH.

Quick Overview for XDH ELISA Kit (ABIN6222583)

Target: XDH (Xanthine Dehydrogenase (XDH))

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 0.16 ng/mL - 10 ng/mL

Application: ELISA

Sample Type: Plasma, Serum

Product Details XDH ELISA Kit

Minimum Detection Limit: 0.16 ng/mL

Purpose: This kit is based on sandwich enzyme-linked immuno-sorbent assay technology.

Analytical Method: Quantitative

Sensitivity: 0.1 ng/mL

Components: The kit components listed are for reference only. The product manual may differ slightly. The product should be used as stated on the product manual included and delivered together with the product.

• Pre-coated 96-Well Microplate
• Standard
• Standard Diluent Buffer
• Wash Buffer
• Detection Reagent A
• Detection Reagent B
• Diluent A
• Diluent B
• TMB Substrate
• Stop Solution
• Plate Sealer

Material not included:

• 37 °C incubator
• Multi and single channel pipettes and sterile pipette tips
• Squirt bottle or automated microplate washer
• 1.5 mL tubes
• Distilled water
• Absorbent filter papers
• 100 mL and 1 liter graduated cylinders
• Microplate reader (wavelength: 450 nm)
• ELISA Shaker

Application Details

Sample Volume: 100 µL

Plate: Pre-coated

Protocol: This kit is based on sandwich enzyme-linked immuno-sorbent assay technology. An antibody is pre-coated onto a 96-well plate. Standards, test samples, and biotin-conjugated reagent are added to the wells and incubated. The HRP-conjugated reagent is then added, and the whole plate is incubated. Unbound conjugates are removed using wash buffer at each stage. TMB substrate is used to quantify the HRP enzymatic reaction. After TMB substrate is added, only wells that contain sufficient XOD will produce a blue coloured product, which then changes to yellow after adding the acidic stop solution. The intensity of the yellow colour is proportional to the XOD amount bound on the plate. The Optical Density (OD) is measured spectrophotometrically at 450 nm in a microplate reader, from which the concentration of XOD can be calculated.

Reagent Preparation:

This procedure is provided for reference only. The product manual may differ slightly. The product should be used as stated on the product manual included and delivered together with the product.

• 1) Standard: Prepare the standard with the recommended volume of Standard Diluent Buffer, to make the standard solution. Then use the Standard Diluent buffer to carry out serial dilutions of the standard solution, as instructed in the Protocol.

• 2) Wash Buffer: Dilute the concentrated Wash Buffer with distilled water, as instructed in the Protocol.

• 3) Detection Reagent Preparation: Calculate the total volume of working solution required. Dilute Detection Reagent A and Detection Reagent B with Diluent A and Diluent B, respectively, at 1:100.

Assay Procedure:

This procedure is provided for reference only. The product manual may differ slightly. The product should be used as stated on the product manual included and delivered together with the product.

• 1) Set standard, test samples and control wells.

• 2) Aliquot 100 μL of diluted standard into the standard wells.

• 3) Aliquot 100 μL of Standard Diluent buffer into control (zero) well.

• 4) Aliquot 100 μL of diluted samples into the sample wells. Incubate for 90 mins at 37 °C.

• 5) Aliquot 100 μL of Detection Reagent A to each well. Incubate for 1 hr at 37 °C.

• 6) Wash 3 times.

• 7) Aliquot 100 μL of Detection Reagent B to each well. Incubate for 30 mins at 37 °C.

• 8) Wash 5 times.

• 9) Aliquot 90 μL of TMB Substrate to each well. Incubate for 10-20 mins at 37 °C.

• 10) Aliquot 50 μL of Stop Solution.

• 11) Measure the OD at 450 nm.

Calculation of Results:

For calculation, average the O.D.450 duplicate readings for each reference standard and each sample and substract the average control (zero) O.D.450 reading. The standard curve can be plotted as the relative O.D.450 of each reference standard solution (Y) vs. the respective concentration of each standard solution (X). The XOD concentration of the samples can be interpolated from the standard curve.

Assay Precision: Intra-assay Precision (Precision within an assay): 3 samples with low, medium and high levels of Xanthine Oxidase (XOD) were were tested 20 times on one plate, respectively.

Inter-assay Precision (Precision between assays): 3 samples with low, medium and high levels of Xanthine Oxidase (XOD) were tested on 3 different plates, 8 replicates in each plate.

CV (%) = (Standard Deviation / mean) x 100

Intra-Assay: CV<10%

Inter-Assay: CV<10%

Restrictions: For Research Use only

Handling

Storage: 4 °C

Storage Comment: Shipped at 4°C. Upon receipt, store the kit according to the storage instruction in the kit's manual.The stability of the kit is determined by the rate of activity loss. The loss rate is less than 5% within the expiration date under appropriate storage conditions. To minimize performance fluctuations, operation procedures and lab conditions should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same user throughout.

Expiry Date: 6 months

Target Details for XDH

Target: XDH (Xanthine Dehydrogenase (XDH))

Alternative Name: Xanthine Dehydrogenase

UniProt: P47989

Pathways: Positive Regulation of Endopeptidase Activity