Ustekinumab (STELARA) ELISA Assay Kit
The Ustekinumab (STELARA) ELISA Assay Kit is For Research Use Only
Size: 12 x 8 wells
Sensitivity: 2.5 ng/mL
Standard Range: 0 – 1000 ng/mL
Incubation Time: 1 hour 10 minutes
Sample Type: Serum, Plasma
Sample Size: 10 µL
Alternative Name: Stelara
Assay Background
Ustekinumab is a human immunoglobulin (Ig) Gl kappa monoclonal antibody directed against interleukin {IL)-12 and IL-23. It was generated via recombinant human IL-12 immunization of human Ig (hu-Ig) transgenic mice. It is a targeted biologic disease-modifying anti-rheumatic drug (bDMARDs) that is used in the management of various inflammatory conditions that involve the activation of IL-12 and IL-23 signaling pathways. Interleukin (IL)-12 and IL-23 are heterodimeric cytokines that evoke immune and inflammatory responses, such as natural killer cell activation and CD4+ T-cell differentiation and activation. The role of IL-12 and IL-23 were implicated in a variety of chronic inflammatory conditions, such as psoriasis and inflammatory bowel diseases. They modulate lymphocyte function, including T-helper (Th) l and Th 17 cell subsets, as CD4+ T cells can differentiate into T-helper (Th) effector lineages based on the environment. Th cells can further activate the downstream pro-inflammatory mediators and transcription factors such as TN Fa and IFNy that drive innate and adaptive immunity. IL-12 and IL-23 share a common p40subunit, paired with p35 and pl9 subunits of IL-12 and IL-23, respectively. The antigen-binding fragment (Fab) of ustekinumab binds the Dl domain of the p40 subunit ofll-12 and IL-23 in al :l ratio. This prevents IL-12 and IL-23 from binding to the IL-12RB1 receptor chain of IL-12 {IL-12RB1/B2) and IL-23 {IL- 12RB1/23R) receptor complexes on the surface of NK and T cells. Ustekinumab only binds to IL-12 and IL-23 that are unbound to IL-12RB1, so it is unlikely to initiate Fe effector functions, such as ADCC or CDC. Inhibition of the IL-12/23 signaling pathway leads to profound suppression of both the Thl and Th 17 cell lineage of cytokines and chemokines and their inflammatory pathways.
Assay Principle
Solid phase enzyme-linked immunosorbent assay {ELISA) based on the sandwich principle. Standards and samples {serum or plasma) are incubated in the microtiter plate coated with the reactant for ustekinumab. After incubation, the wells are washed. Then, horse radish peroxidase {HRP) conjugated probe is added and binds to ustekinumab captured by the reactant on the surface of the wells. Following incubation wells are washed and the bound enzymatic activity is detected by addition of tetramethylbenzidine {TMB) chromogen substrate. Finally, the reaction is terminated with an acidic stop solution. The color developed is proportional to the amount of ustekinumab in the sample or standard. Results of samples can be determined directly using the standard curve.
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