Infliximab ELISA Kit
-
- Target
- Infliximab
- Reactivity
- Human, Mouse, Rat, Monkey
- Detection Method
- Colorimetric
- Method Type
- Sandwich ELISA
- Detection Range
- 0.2-6 μg/mL
- Minimum Detection Limit
- 0.2 μg/mL
- Application
- ELISA
- Purpose
- Enzyme immunoassay for the quantitative determination of free Infliximab in human serum and plasma.
- Brand
- ImmunoGuide®
- Sample Type
- Plasma, Serum
- Analytical Method
- Quantitative
- Specificity
- Infliximab (Remicade®, Remsima®)
- Sensitivity
- 10 ng/mL
- Characteristics
- This kit measures the free drug concentration.
- Components
- plate, standards, assay buffer, conjugate, TMB, HCl, wash buffer
- Material not included
- Normal lab equipment for performing ELISA assays
-
- Application Notes
-
- Before performing the assay, samples and assay kit should be brought to room temperature (about 30 minutes beforehand) and ensure the homogeneity of the solution.
- All Standards should be run with each series of unknown samples.
- Standards should be subject to the same manipulations and incubation times as the samples being tested.
- All steps of the test should be completed without interruption. 6
- Use new disposable plastic pipette tips for each reagent, standard or specimen in order to avoid cross contamination.
- Comment
-
0.2 - 6 µg/mL (in case of the recommended sample dilution factor)
10 ng/ml (sensitivity in the well) - Sample Volume
- 20 μL
- Assay Time
- 1.5 h
- Plate
- Pre-coated
- Protocol
- This is a sandwich type ELISA. Standards and diluted samples (serum or plasma) are incubated in the microtitre plate coated with recombinant human TNF-a (rh TNF-a). After incubation, the wells are washed. A horseradish peroxidase (HRP)-conjugated anti-human IgG monoclonal antibody is added and binds to the Fc part of Infliximab pre-captured by the rh TNF-a on the surface of the wells. Following incubation, the wells are washed and the bound enzymatic activity is detected by addition of chromogen-substrate. The colour developed is proportional to the amount of Infliximab in the sample or standard. Results of samples can be determined directly by using the standard curve.
- Reagent Preparation
-
Just the wash buffer has to be prepared by diluting the stock solution 1:20. All other reagents are ready to use.
- Sample Collection
- Normal serum or plasma collection
- Sample Preparation
-
dilute the samples 1:20 with assay buffer
- Assay Procedure
-
Pipette 100 µL of assay buffer into each of the wells.
Pipette 20 µL of each standard and diluted samples.
Incubate 30 min at room temperature (RT).
Aspirate or decant the incubation solution.
Wash the plate 3 x 300 μL of diluted wash buffer per well.
Pipette 100 μL of enzyme conjugate (HRP-anti human IgG mAb).
Shake plate carefully. Incubate 30 min at RT.
Wash the plate 3 x 300 μL of diluted wash buffer per well.
Pipette 100 µL TMB substrate solution into each well.
Incubate 15 min at RT.
Add 100 µL of stop solution.
Measure optical density (OD) at 450 nm. - Calculation of Results
-
The results are read from a standard curve.
- Assay Precision
- < 10%
- Restrictions
- For Research Use only
-
- Buffer
- < 15mM NaN3
- Preservative
- Sodium azide
- Precaution of Use
- This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
- Storage
- 4 °C
- Storage Comment
-
The kit is shipped at ambient temperature and should be stored at 2-8°C.
Keep away from heat or direct sun light.
The storage and stability of specimen and prepared reagents is stated in the corresponding chapters.
The microtiter strips are stable up to the expiry date of the kit in the broken, but tightly closed bag when stored at 2-8°C. - Expiry Date
- 24 months
-
- Target
- Infliximab
- Target Type
- Antibody
- Background
- The drug Infliximab (trade name Remicade® and Remsima®) is a chimeric monoclonal antibody and used to treat auto-immune disorders
- Molecular Weight
- 144 kDa
-
