ELISA/assay

Total OKT3 ELISA Kit

DEIA-JY25043

96T

Total OKT3 ELISA Kit

CD3e

Total OKT3 ELISA Kit

ELISA

CD3e

Human

Quantitative

Plasma, Serum

96T

2-8°C

Human

This kit is designed for the quantitative detection of total

Solid phase enzyme-linked immunosorbent assay (ELISA) based on the sandwich principle. Standards and samples (serum or plasma) are incubated in the microtiter plate coated with the reactant for

1. Pre-coated Microplate: 1 plate 2-8°C 2. Standards: 6 vials, -20°C 3. Sample Diluent: 15 mL, 2-8°C 4. HRP-Conjugated Antibody: 10 mL, 2-8°C 5. TMB Substrate: 10 mL, 2-8°C 6. Stop Solution: 10 mL, RT 7. Wash Buffer (20X): 30 mL, RT 8. Plate Sealer: 2 sheets, RT

1. Micropipettes and tips 2. Calibrated measures 3. Tubes for sample dilution 4. Wash bottle, automated or semi-automated microtiter plate washing system 5. Microtiter plate reader capable of measuring optical density with a photometer at OD 450 nm with reference wavelength 650 nm (450/650 nm) 6. Distilled or deionised water, paper towels, pipette tips and timer

The usual precautions for venipuncture should be observed. Do not use grossly haemolytic, icteric or lipemic specimens. Samples appearing turbid should be centrifuged before testing to remove any particulate material. Avoid repeated freezethaw cycles for serum/plasma samples. Samples should be diluted with the dilution rate given in the "Reagent Preparation" before the test. Drug infusions may camouflages/mask the presence of antibody to drugs in serum/plasma samples. Therefore, blood sampling time is critical for detection of antibodies. It is recommended to take the blood sample just before the scheduled dose (trough specimen). Stability (serum/plasma): 2-8°C, 2 days -20°C, 6 months

Procedure Notes: 1. Any improper handling of samples or modification of the test procedure may influence the results. The indicated pipetting volumes, incubation times, temperatures and pre-treatment steps must be performed strictly according to the instructions. Use calibrated pipettes and devices only. 2. Once the test has been started, all steps should be completed without interruption. Make sure that required reagents, materials and devices are prepared ready at the appropriate time. Allow all reagents and specimens to reach room temperature (18-25°C) and gently swirl each vial of liquid reagent and sample before use. Mix reagents without foaming. 3. Avoid contamination of reagents, pipettes and wells/tubes. Use new disposable plastic pipette tips for each reagent, standard or specimen. Do not interchange caps. Always cap not used vials. Do not reuse wells/tubes or reagents. 4. Use a pipetting scheme to verify an appropriate plate layout. 5. Incubation time affects results. All wells should be handled in the same order and time sequences. It is recommended to use an eight-channel micropipette for pipetting of solutions in all wells. 6. Microplate washing is important. Improperly washed wells will give erroneous results. It is recommended to use a multichannel pipette or an automatic microplate washing system. Do not allow the wells to dry between incubations. Do not scratch coated wells during rinsing and aspiration. Rinse and fill all reagents with care. While rinsing, check that all wells are filled precisely with wash buffer, and that there are no residues in the wells. 7. Humidity affects the coated wells/tubes. Do not open the pouch until it reaches room temperature. Unused wells/tubes should be returned immediately to the resealed pouch including the desiccant. Component: Wash buffer (Must be prepared before starting assay procedure) Dilute: 10 mL (e.g.) With Distilled water: Up to 200 mL Dilution Ratio: 1/20 Remarks: Warm up 37°C to dissolve crystals. Mix vigorously Storage: 2-8°C Stability: 2 weeks Dilution of samples Sample: Serum/Plasma Diluent: Assay buffer Dilution Ratio: 1/100 Remarks: Dilution 1/100, 5 μL sample + 495 μL assay buffer Patient samples with a concentration of drug above the measuring range are to be rated as > "Highest Standard (Standard A)". The result must not be extrapolated. The patient sample in question should be further diluted with assay buffer and retested.

Total assay time: 105 minutes 1. Pipette 100 μL of each "Standards" and diluted samples into the respective wells of microtiter plate Wells A1: Standard A B1: Standard B C1: Standard C D1: Standard D E1: Standard E F1: Standard F 2. - Cover the plate with adhesive foil - Briefly mix contents by gently shaking the plate - Incubate 60 minutes at room temperature (18-25°C) 3. - Remove adhesive foil - Discard incubation solution - Wash plate three times each with 300 μL "Wash Buffer" - Remove excess solution by tapping the inverted plate on a paper towel 4. Pipette 100 μL "Conjugate" into each well 5. - Cover the plate with adhesive foil - Incubate 30 minutes at room temperature (18-25°C) 6. - Remove adhesive foil - Discard incubation solution - Wash plate three times each with 300 μL "Wash Buffer" - Remove excess solution by tapping the inverted plate on paper towel 7. - Pipette 100 μL "Substrate" into each well 8. - Incubate 15 minutes without adhesive foil at room temperature (18-25°C) in the dark 9. - Stop the substrate reaction by adding 100 μL "Stop Solution" into each well. Briefly mix contents by gently shaking the plate Colour changes from blue to yellow 10. Measure optical density with a photometer at OD 450 nm with reference wavelength 650 nm (450/650 nm) within 30 minutes after pipetting the "Stop Solution

For a valid study, the OD 450/650 of the highest standard should be> 1.500 and the OD 450/650 of the lowest standard should be Expiration dates of reagents, storage conditions, pipettes, devices, incubation conditions, washing methods, etc.

1. Create a standard curve by using the standards. OD 450/650 nm for each standard on the vertical (Y-axis) axis versus the corresponding drug concentration on the horizontal (X-axis) axis. 2. The concentration of the samples can be read directly from this standard curve. Using the absorbance value for each sample, determine the corresponding concentration of drug from the standard curve. Find the absorbance value on the Y-axis and extend a horizontal line to the curve. At the point of intersection, extend a vertical line to the X-axis and read the drug concentration of the unknown sample. 3. If computer data is going to be used, we recommend primarily "Four Parameter Logistic (4PL)" or secondly the "point-to-point calculation". 4. To obtain the exact values of the samples, the concentration determined from the standard-curve must be multiplied by the dilution factor (10×). Any sample reading greater than the highest standard should be further diluted appropriately with assay buffer and retested. Therefore, if the pre-diluted samples have been further diluted, the concentration determined from the standard curve must be multiplied by the further dilution factor.

Intra-assay and inter-assay CVs < 30%

< 100 ± 30%

1. For professional use only. 2. Obey lot number and expiry date. Do not mix reagents of different lots. Do not use expired reagents. 3. Before starting the assay, read the instructions completely and carefully. Use the valid version of the package insert provided with the kit. Be sure that everything is understood. For further information (clinical background, test performance, automation protocols, alternative applications, literature, etc.) please refer to the local distributor. 4. Follow good laboratory practice and safety guidelines. Wear lab coats, disposable latex gloves and protective glasses where necessary. 5. Reagents of this kit containing hazardous material may cause eye and skin irritations. 6. Chemicals and prepared or used reagents must be treated as hazardous waste according the national biohazard safety guidelines or regulations.

This kit is designed for the quantitative detection of total

1. Pre-coated Microplate: 1 plate 2-8°C 2. Standards: 6 vials, -20°C 3. Sample Diluent: 15 mL, 2-8°C 4. HRP-Conjugated Antibody: 10 mL, 2-8°C 5. TMB Substrate: 10 mL, 2-8°C 6. Stop Solution: 10 mL, RT 7. Wash Buffer (20X): 30 mL, RT 8. Plate Sealer: 2 sheets, RT

See the individual product datasheet

See the individual product datasheet

See the individual product datasheet

See the individual product datasheet

See the individual product datasheet

Solid phase enzyme-linked immunosorbent assay (ELISA) based on the sandwich principle. Standards and samples (serum or plasma) are incubated in the microtiter plate coated with the reactant for