Anti-Omalizumab ELISA Assay Kit

$1,725.00

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

SKU: KBI2021 Categories: , ,

Anti-Omalizumab ELISA Assay Kit

The Anti-Omalizumab 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
Alternative Name: Xolair

Assay Background

Omalizumab, sold under the trade name Xolair, is a medication originally designed to reduce sensitivity to allergens. It has been used to try to control severe allergic asthma, which does not respond to high doses of corticosteroids and less widely for chronic spontaneous urticaria. Omalizumab is a recombinant DNA-derived humanized IgG1k monoclonal antibody that specifically binds to free human immunoglobulin E (IgE) in the blood and interstitial fluid and to membrane-bound form of IgE (mIgE) on the surface of mIgE-expressing B lymphocytes. Unlike an ordinary anti-IgE antibody, it does not bind to IgE that is already bound by the high affinity IgE receptor (FcεRI) on the surface of mast cells, basophils, and antigen-presenting dendritic cells. 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 Omalizumab and can be responsible for allergic reaction, or even anaphylactic shock. This ELISA kit detects antibodies for Anti-Omalizumab and may be used for monitoring immunogenicity.

Related Products

Omalizumab ELISA Assay Kit
Omalizumab mAb-Based ELISA Assay
Omalizumab IgE ELISA Assay

Additional Information

Assay Principle

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

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