Ustekinumab ELISA Assay Kit

$1,790.00

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

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Ustekinumab ELISA Assay Kit

The Ustekinumab ELISA Assay Kit is For Research Use Only

Size: 12×8 wells
Sensitivity: <5 ng/mL
Standard Range: 5-320 ng/ml
Incubation Time: 2.5 hours
Sample Type: Serum, Plasma, Cell Culture Supernatants
Sample Size: 100 µL
Alternative Name: Stelara


Assay Background for Ustekinumab ELISA Assay

Ustekinumab is a human monoclonal antibody. It is directed against interleukin 12and interleukin 23, naturally occurring proteins that regulate the immune system and immune-mediated inflammatory disorders. In two Phase III trials for moderate to severe psoriasis, the longest >76 weeks, ustekinumab was safe and effective. A third Phase III trial, ACCEPT, compared the efficacy and safety of ustekinumab with etanercept in the treatment of moderate to severe plaque psoriasis. This trial found a significantly higher clinical response with ustekinumab over the 12-week study period compared to high-dose etanercept. It also demonstrated the clinical benefit of ustekinumab among patients who failed to respond to etanercept. Ustekinumab is approved in Canada, Europe and the United States to treat plaque psoriasis.


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.


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Additional Information

Assay Principle


The method employs the quantitative sandwich enzyme immunoassay technique. Antibodies to Ustekinumab are pre-coated onto microwells. Samples and standards are pipetted into microwells and human Ustekinumab present in the sample are bound by the capture antibody. Then, a HRP (horseradish peroxidase) conjugated anti-Ustekinumab 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 Ustekinumab 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 25 μl of Assay Diluent in each well.
3. Pipette 25 μl of Anti-Ustekinumab: HRP Conjugate into each well.
4. Add 100 μl of Standards or Samples into the respective wells.
5. Cover the plate and incubate for 120 minutes at 37°C
6. 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.
7. Add 100 μl of TMB Substrate in each well.
8. 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.
9. Pipette out 100 μl of Stop Solution. Wells should turn from blue to yellow in color.
10. Read the absorbance at 450 nm with a microplate reader.

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