TSH ELISA Kit

Catalog No. ABIN6227249

Mouse TSH ELISA Kit, Colorimetric assay for quantification of Mouse TSH.

Quick Overview for TSH ELISA Kit (ABIN6227249)

Target: TSH (Thyroid Stimulating Hormone (TSH))

Reactivity: Mouse

Detection Method: Colorimetric

Method Type: Competition ELISA

Detection Range: 6.4 pg/mL - 4000 pg/mL

Application: ELISA

Sample Type: Plasma, Serum

Grade: Small Sample

Product Details TSH ELISA Kit

Minimum Detection Limit: 6.4 pg/mL

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

Analytical Method: Quantitative

Sensitivity: < 2.4 pg/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: 25 µL

Plate: Pre-coated

Protocol: This kit is based on competitive 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. A competitive inhibition reaction takes place between the biotin-labelled TSH and the unlabelled- TSH on the pre-coated antibody. 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 TSH will produce a blue coloured product, which then changes to yellow after adding the acidic stop solution. The intensity of the color yellow is inversely proportional to the TSH amount bound on the plate. The OD is measured spectrophotometrically at 450 nm in a microplate reader, from which the concentration of TSH 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 25 μL of diluted standard into the standard wells.

• 3) Aliquot 25 μL of Standard Diluent buffer into the control (zero) well.

• 4) Aliquot 25 μL of diluted samples into the sample wells.

• 5) Immediately aliquot 25 μL of Detection Reagent A to each well. Incubate for 1 hr at 37 °C.

• 6) Wash 3 times.

• 7) Aliquot 50 μL of Detection Reagent B to each well. Incubate for 30 mins at 37 °C.

• 8) Wash 5 times.

• 9) Aliquot 50 μL of TMB Substrate to each well. Incubate for 10-20 mins at 37 °C.

• 10) Aliquot 25 μL of Stop Solution.

• 11) Measure the OD at 450 nm.

Calculation of Results:

This assay is competitive, therefore there is an inverse correlation between TSH concentration in the sample and the absorbance measured. Create a graph with the log of the standard concentration (y-axis) and average absorbance measured (x-axis). Apply a best fit trendline through the standard points. The TSH 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 Thyroid Stimulating Hormone (TSH) were were tested 20 times on one plate, respectively.

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

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

Intra-Assay: CV<10%

Inter-Assay: CV<12%

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 TSH

Target: TSH (Thyroid Stimulating Hormone (TSH))

Alternative Name: Thyroid Stimulating Hormone