Therapeutic Drug Monitoring ELISA Assay

Ustekinumab mAb-based ELISA Assay

$1,485.00

The Eagle Biosciences Ustekinumab ELISA Assay Kit (mAb-based) is an enzyme immunoassay for the specific determination of free Ustekinumab in serum and plasma. The Ustekinumab ELISA (maB-based) Assay Kit is for research use only and not to be used in diagnostic procedures.

SKU: IG-AB121 Categories: , , Tag:

Ustekinumab mAb-based ELISA Assay

The Ustekinumab mAb-based ELISA Assay is For Research Use Only

Size: 1 x 96 wells
Sensitivity: 1.5 ng/mL
Dynamic Range: 2-60 ng/mL
Incubation Time: 2.5 hours
Sample Type: Serum, Plasma
Sample Size: 10 µL
Alternative Name: Stelara

Assay Principle

The Ustekinumab mAb-based ELISA is based on Ustekinumab-specific monoclonal antibody (catcher Ab, ImmunoGuide clone 9C7). Diluted standards and samples are incubated in the microtiter plate coated with IG-9C7 mAb. After incubation, the wells are washed. A biotinylated anti-human IgG monoclonal antibody (clone IG-1B5; specific for the Fc part of all human IgG i.e. IgG1, IgG2, IgG3 and IgG4) is added and binds to the Fc part of Ustekinumab. Following incubation, wells are washed and the horseradish peroxidase (HRP)-conjugated streptavidin is added and binds to the biotinylated 1B5 mAb.  Following incubation, wells are washed and the bound enzymatic activity is detected by addition of chromogen-substrate. The color developed is proportional to the amount of Ustekinumab in the sample or standard. Results of samples can be determined by using the standard curve. Preincubation of Ustekinumab with recombinant human interleukin-12 (IL-12), contains p40 protein subunit as that of IL-23, inhibited the reaction. Therefore, the Eagle Biosciences Ustekinumab ELISA (mAb-based) measures the free form of Ustekinumab.

Storage and Stability of the Ustekinumab mAb-based ELISA Assay
The kit is shipped at ambient temperature and should be stored at 2-8°C. Keep away from heat or direct sun light. 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.

Related Products

Ranibizumab mAb-based ELISA Assay Kit

Additional Information

Assay Background

The drug Ustekinumab (trade name Stelara®) is a humanized immunoglobulin G1k monoclonal antibody that binds with high specificity to the p40 protein subunit used by both the IL-12 and IL-23 cytokines. The ImmunoGuide Ustekinumab ELISA (mAb-based) kit can be efficiently used for measuring free Certulizumab pegol levels in serum and plasma.

Assay Procedure

  1. Pipette 100 µL of Assay Buffer into each of the wells to be used.
  2. Pipette 75 µL of each 1:10 Diluted Standard, and 1:200 Diluted Samples into the respective wells of the microtiter plate. Bubble formation during the pipetting of standards and samples must be avoided.
  3. Cover the plate with adhesive seal. Shake plate carefully by tapping several times. Incubate the plate on bench top for 60 min at room temperature (RT, 20-25°C).
  4. Remove adhesive seal. Aspirate or decant the incubation solution. Wash the plate 3 X 300 μL of Diluted Wash Buffer per well. Remove excess solution by tapping the inverted plate on a paper towel.
  5. Pipette 100 μL of Biotinylated a-hIgG into each well.
  6. Cover plate with adhesive seal. Shake plate carefully by tapping several times. Incubate the plate on a bench top for 30 min at RT.
  7. Remove adhesive seal. Aspirate or decant the incubation solution. Wash the plate 3 X 300 μL of Diluted Wash Buffer per well. Remove excess solution by tapping the inverted plate on a paper towel.
  8. Pipette 100 μL of Enzyme Conjugate (HRP-Streptavidin) into each well.
  9. Cover plate with adhesive seal. Shake plate carefully by tapping several times. Incubate the plate on a bench top for 30 min at RT.
  10. Remove adhesive seal. Aspirate or decant the incubation solution. Wash the plate 3 X 300 μL of Diluted Wash Buffer per well. Remove excess solution by tapping the inverted plate on a paper towel.
  11. Pipette 100 µL of Ready-to-Use TMB Substrate Solution into each well.
  12. Incubate 20 min at RT. Avoid exposure to direct sunlight.
  13. Stop the substrate reaction by adding 100 µL of Stop Solution into each well. Color changes from blue to yellow. Briefly mix contents by gently shaking the plate.
  14. Measure optical density (OD) with a photometer at 450 nm (Reference at OD620 nm is optional) within 15 min after pipetting the Stop Solution.

Typical Calibration Curve

Documents

Product Manual

 

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.

Product Citations