KBI2022 Anti-Tocilizumab ELISA Assay Kit

Anti-Tocilizumab ELISA Assay Kit

$1,810.00

The Anti-Tocilizumab ELISA Assay Kit is used as an analytical tool for quantitative determination of Anti-Tocilizumab in serum, plasma and cell culture supernatant.

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Anti-Tocilizumab ELISA Assay Kit

The Anti-Tocilizumab ELISA Assay Kit is For Research Use Only

Size: 12×8 wells
Sensitivity: <10 ng/mL
Standard Range: 10-640 ng/ml
Incubation Time: 2.25 hours
Sample Type: Serum, Plasma, Cell Culture Supernatants
Sample Size: 100 µL
Alternate Name: Actemra

Assay Background

Tocilizumab, also known as atlizumab, is an immunosuppressive drug, mainly for the treatment of rheumatoid arthritis (RA) and systemic juvenile idiopathic arthritis, a severe form of arthritis in children. It is a humanized monoclonal antibody against the interleukin-6 receptor (IL-6R). Interleukin 6 (IL-6) is a cytokine that plays an important role in immune response and is implicated in the pathogenesis of many diseases, such as autoimmune diseases, multiple myeloma and prostate cancer. It was developed by Hoffmann–La Roche and Chugai. Anti-Drug Antibodies (ADA) may induce unwanted side effects in biopharmaceuticals. Hence, ADA has been subjected to increase in scrutiny by the regulatory authorities using immunogenicity safety studies. ADA has been observed in pre-clinical and clinical studies, resulting in significant changes in toxicology, pharmacokinetics and efficacy. These effects result from the generation of drug-induced (neutralizing) autoantibodies against Tocilizumab and can be responsible for allergic reaction, or even anaphylactic shock. This ELISA kit detects antibodies for Anti-Tocilizumab and may be used for monitoring immunogenicity.

Related Products

Tocilizumab ELISA Assay Kit
Tocilizumab mAb-based ELISA Assay
Anti-Eculizumab (Soliris) ELISA Assay Kit

Additional Information

Assay Principle

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

Assay Procedure

  1. It is strongly recommended that all Controls and Samples be run in duplicates or triplicates. A standard curve is required for each assay. All steps must be performed at 37°C.
  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 37°C.
  5. 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.
  6. Pipette without delay in the same order 100 μl of Tocilizumab: HRP Conjugate into each well.
  7. Cover the plate and incubate for 60 minutes at 37°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 37°C for 15 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.

Package Inserts

Please note: All documents above are for reference use only and should not be used in place of the documents included with this physical product. If digital copies are needed, please contact us.

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