Description

Datasheet

Ustekinumab (Stelara®) ELISA Kit

Enzyme-linked immunosorbent assay (ELISA) for the quantitative determination of Ustekinumab (Stelara®) in human serum and plasma.This Assay Genie kit has been especially developed for the quantitative analysis of Ustekinumab in serum and plasma samples.

Ustekinumab (Stelara®) ELISA Kit test principle

Solid phase enzyme-linked immunosorbent assay (ELISA) based on the sandwich principle. Standards and samples (serum or plasma) are incubated in the microtitre plate coated with the reactant for Ustekinumab (Stelara®). After incubation, the wells are washed. A 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 chromogen-substrate. The colour developed is proportional to the amount of ustekinumab in the sample or standard. Results of samples can be determined directly using the standard curve.

Ustekinumab (Stelara®) Product Information

Information	Description
Application	Free drug
Required Volume (uL)	10
Total Time (min)	70
Sample Type	Serum, Plasma
Number of Assays	96
Detection Limit (ng/mL)	3 (ng/mL)
Spike Recovery (%)	85- 115%
Shelf Life (year)	1
Alternative Names	Stelara

Ustekinumab (Stelara®) - Key Information

Ustekinumab (Stelara®) mode of action

Ustekinumab is a fully human monoclonal antibody that binds with high specificity and affinity to the cytokines interleukin (IL)-12 and IL-23, thereby suppressing IL-12- and IL-23-mediated inflammation associated with psoriasis. Ustekinumab is a human IgG1-kappa monoclonal antibody designed to interfere with the triggering of the body's inflammatory response through the suppression of certain cytokines. Specifically, ustekinumab blocks interleukin IL-12 and IL-23 by binding with high affinity and specificity to the p40 subunit of IL-12 and IL-23, therefore disrupting the proinflammatory pathway that contributes to several chronic diseases. In a more minor role, ustekinumab also interferes with the expression of monocyte chemotactic protein-1 (MCP-1), tumor necrosis factor-alpha (TNF-alpha), interferon-inducible protein-10 (IP-10), and interleukin IL-8.

Ustekinumab (Stelara®) uses

Ustekinumab is used as a treatment for plaque psoriasis and a particular type of arthritis (psoriatic arthritis). Furthermore, ustekinumab is used for certain bowel conditions including Crohn's disease and ulcerative colitis.

Ustekinumab (Stelara®) treatment

Ustekinumab (STELARA®) is indicated for the treatment of adult patients with moderate to severe plaque psoriasis (Ps) who are candidates for phototherapy or systemic therapy. It works by blocking certain natural proteins in your body (interleukin-12 and interleukin-23) that cause inflammation (swelling) in these conditions. Ustekinumab (STELARA®) is the only FDA-approved medicine that targets IL-12 and IL-23, which are thought to be associated with gastrointestinal inflammation in Crohn's disease.

Ustekinumab (Stelara®) ELISA Kit Contents

Size	Kit Contents
1 x 12 x 8	Microtiter Plate Break apart strips. Microtiter plate with 12 rows each of 8 wells coated with reactant
7 x 0.2 mL	Ustekinumab Standards A-E (10X), High Level Control (10X), Low Level Control (10x) 1000; 300; 100; 30; 0 nanogram/mL Used for construction of the standard curve. C
1 x 50 mL	Assay Buffer Blue coloured. Ready to use. Contains proteins and <0.1% NaN3.
1 x 12 mL	Horse radish peroxidase-Conjugated Probe. Red coloured. Ready to use. Contains HRP-probe, stabilizer and <0.1% NaN3.
1 x 12 mL	TMB Substrate Solution Ready to use. Contains TMB
1 x 12 mL	TMB Stop Solution Ready to use. 1N HCl
1 x 50 mL	Wash Buffer concentrate (20x) Contains Buffer with Tween 20.
2 x 1	Adhesive Foil For covering of Microtiter Plate during incubation.

Ustekinumab (Stelara®) ELISA Protocol

Steps	Protocol
1	Dilute each of the standards and samples (serum/plasma) using Assay Buffer as described in “Dilution of Standards and Samples (serum/plasma)” section
2	Pipette 75µl of Assay Buffer non-exceptionally into each of the wells to be used.
3	Pipette 25 µL of each Diluted Standards, Diluted High Level Control, Low Level Control and Diluted Samples into the respective wells of microtiter plate. Wells A1: Diluted Standard A B1: Diluted Standard B C1: Diluted Standard C D1: Diluted Standard D E1: Diluted Standard E F1: High Level Control G1: Low Level Control H1 and on: Diluted Sample (Serum/Plasma)
4	Cover the plate with adhesive foil. Incubate 30 min at room temperature (18- 25°C).
5	Remove adhesive foil. Discard incubation solution. Wash plate 3 times each with 300 µL of diluted Wash Buffer. Remove excess solution by tapping the inverted plate on a paper towel
6	Pipette 100 µL of ready-to use HRP-Conjugated Probe into each well
7	Cover the plate with adhesive foil. Incubate 30 min at room temperature (18- 25°C).
8	Remove adhesive foil. Discard incubation solution. Wash plate 3 times each with 300 µL of diluted Wash Buffer. Remove excess solution by tapping the inverted plate on a paper towel.
9	Pipette 100 µL of TMB Substrate Solution into each well.
10	Incubate 10 min (without adhesive foil.) at room temperature (18-25°C) in the dark.
11	Stop the substrate reaction by adding 100 µL of Stop Solution into each well. Briefly mix contents by gently shaking the plate. Colour changes from blue to yellow
12	Measure optical density with a photometer at 450/650 nm within 30 min after pipetting of the Stop Solution.