Infliximab ELISA Assay Kit

$955.00

The Infliximab ELISA Assay Kit is intended as an analytical tool for quantitative determination of Infliximab in serum, plasma and cell culture supernatant.

SKU: KBI1011 Categories: , , Tag:

Infliximab ELISA Assay Kit

The Infliximab ELISA Assay Kit is For Research Use Only

Size: 12×8 wells
Sensitivity: <2.5 ng/mL
Standard Range: 2.5-160 ng/ml
Incubation Time: 1.5 hours
Sample Type: Serum, Plasma, Cell Culture Supernatants
Sample Size: 100 µL
Alternative Name: Remicade


Assay Principle

The method employs the quantitative sandwich enzyme immunoassay technique. Antibodies to Infliximab are pre-coated onto microwells. Samples and standards are pipetted into microwells and human Infliximab present in the sample are bound by the capture antibody. Then, a HRP (horseradish peroxidase) conjugated anti-Infliximab antibody is pipetted and incubated. After washing microwells in order to remove any non-specific binding, the ready to use substrate solution (TMB) is added to microwells and color develops proportionally to the amount of Infliximab in the sample. Color development is then stopped by addition of stop solution. Absorbance is measured at 450 nm.


Sample Preparation and Storage:
Blood is taken by venipuncture. Serum is separated after clotting by centrifugation. Plasma can be used, too. Lipaemic, hemolytic or contaminated samples should not be run. Repeated freezing and thawing should be avoided. If samples are to be used for several assays, initially aliquot samples and keep at – 20°C. For Cell Culture Supernatant – If necessary, centrifuge to remove debris prior to analysis. Samples can be stored at -20°C or -80⁰C. Avoid repeated freeze-thaw cycles.

Preparation Before Use: Allow samples to reach room temperature prior to assay. Take care to agitate patient samples gently in order to ensure homogeneity.
Test Sample preparation – Samples have to be diluted 1:10 to 1:100 (v/v), e.g. for 1:100 (5 µl sample + 495 µl sample diluent) prior to assay. The samples may be kept at 2 – 8°C for up to three days. Long-term storage requires -20°C.


Related Products

Anti-Infliximab (Remicade) ELISA Assay Kit
Total Infliximab ELISA Assay
Infliximab ELISA mAb-based Assay

Additional Information

Assay Background


Infliximab is a monoclonal antibody which has high specificity for TNFα, and does not neutralize TNF beta. Infliximab causes programmed cell death of TNFα-expressing activated T lymphocytes, an important cell type mediating inflammation; hence it is generally assumed that resolution of activated T cells by Infliximab explains its efficacy in Crohn’s disease.

Assay Procedure


  1. It is strongly recommended that all Controls and Samples be run in duplicates or triplicates. A standardcurve is required for each assay. All steps must be performed at room temperature (RT).
  2. Pipette out 50 μl of Assay Diluent in each well.
  3. Add 100 μl of Standards or Samples into the respective wells.
  4. Cover the plate and incubate for 60 minutes at room temperature, 22°C±4°C.
  5. Aspirate and wash plate 4 times with Wash Buffer (1X) and blot residual buffer by firmly tapping plateupside down on absorbent paper. Wipe off any liquid from the bottom outside of the microtiter wells as any residue can interfere in the reading step.
  6. Pipette without delay in the same order 50 μl of Anti-Infliximab: HRP Conjugate into each well.
  7. Cover the plate and incubate for 60 minutes at room temperature, 22°C±4°C.
  8. Aspirate and wash plate 4 times with Wash Buffer (1X) and blot residual buffer by firmly tapping plate upside down on absorbent paper. Wipe off any liquid from the bottom outside of the microtiter wells as any residue can interfere in the reading step.
  9. Add 100 μl of TMB Substrate in each well.
  10. Incubate the plate at room temperature for 30 minutes in dark. DO NOT SHAKE or else it may result in higher backgrounds and worse precision. Positive wells should turn bluish in color.
  11. Pipette out 100 μl of Stop Solution. Wells should turn from blue to yellow in color.
  12. Read the absorbance at 450 nm with a microplate reader.

Documents

Product Documents


Product Citations