Tocilizumab ELISA Assay Kit

$1,155.00

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

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Tocilizumab ELISA Assay Kit

The Tocilizumab 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: 2.5 hours
Sample Type: Serum, Plasma, Cell Culture Supernatants
Sample Size: 100 µL
Alternative Name: Actemra


Assay Principle

The method employs the quantitative sandwich enzyme immunoassay technique. Antibodies to Tocilizumab are pre-coated onto microwells. Samples and standards are pipetted into microwells and human Tocilizumab present in the sample are bound by the capture antibody. Then, a HRP (horseradish peroxidase) conjugated anti-Tocilizumab 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 Tocilizumab 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.


Related Products

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

Additional Information

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.

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 Anti-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 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.

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