Nivolumab ELISA Kit (Opdivo®) Free drug

Product type:
Biosimilar ELISA
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Nivolumab (Opdivo®) ELISA Kit

Enzyme-linked immunosorbent assay (ELISA) for the quantitative determination of nivolumab (Opdivo®) in serum and plasma. Assay Genie Nivolumab ELISA has been specially developed for the quantitative analysis of free nivolumab in serum and plasma samples at high specificity.

Nivolumab (Opdivo®) 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 nivolumab (Opdivo®). After incubation, the wells are washed. A horse radish peroxidase (HRP) conjugated probe is added and binds to nivolumab 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 nivolumab in the sample or standards. Results of samples can be determined directly using the standard curve.

Nivolumab (Opdivo®) Product Information

Information Description
Free drug
Required Volume (uL)
Total Time (min)
Sample Type
Serum, Plasma
Number of Assays
Detection Limit (ng/mL)
10 (ng/mL)
Spike Recovery (%)
Shelf Life (year)

Alternative Names

Human IgG4 monoclonal antibody

Anti-PD-1 mAb


Nivolumab (Opdivo®) - Key Information

Nivolumab (Opdivo®) mode of action

Nivolumab is a fully human IgG4 monoclonal antibody that acts as an immunomodulator by blocking ligand activation of programmed cell death 1 (PD-1) receptor on T cells. Nivolumab binds to the programmed cell death 1 (PD-1) receptor and selectively blocks interaction with its programmed death ligands PD-L1 and PD-L2. Upregulation of PD-1 ligands occurs in some tumors and signalling through this pathway can contribute to inhibition of active T-cell immune surveillance of tumour tissue. The inhibitory effect of PD-1 and its ligands occurs through the promotion of apoptosis in antigen specific T cells while simultaneously blocking apoptosis in suppressor T cells. Blocking PD-1 activity has been shown to lead to decreased tumour growth in mouse tumour models.

Nivolumab (Opdivo®) uses

Nivolumab (Opdivo®) is indicated for use in patients with unresectable (cannot be surgically removed) or metastatic melanoma who no longer respond to other drugs. It is also used to treat metastatic squamous non-small cell lung cancer and is a second-line treatment for renal cell carcinoma after anti-angiogenic treatment has failed.

Nivolumab (Opdivo®) treatment

Nivolumab is administered as an intravenous infusion over 60 minutes every 2 weeks. Nivolumab is indicated for the treatment of unresectable or metastatic melanoma for patients who no longer respond to treatment with other drugs. It is intended for use in patients who have been previously treated with ipilimumab and is used for melanoma patients after treatment with ipilimumab and a BRAF inhibitor in patients whose tumors express BRAF V600 gene mutations. Historically there have been very few effective treatments for advanced melanoma, which is why this product was approved under an FDA accelerated program to allow earlier patient access.

Nivolumab (Opdivo®) 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

8 x 0.3 mL

Nivolumab Standards A-F, High Level Control, Low Level Control (10x)

30; 1; 0.3; 0.1; 0.03; 0 microgram/mL
Used for construction of the standard curve. Contains nivolumab (Opdivo),
human serum, stabilizer and <0.1% NaN3

2 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 preservatives.

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.

Nivolumab (Opdivo®) ELISA Protocol

Steps Protocol


Dilute each of the standards and samples (serum/plasma) using Assay Buffer as described in “Dilution of Standards and Samples (standards/serum/plasma)” section.


Pipette 100µl of Assay Buffer non-exceptionally into each of the wells to be used.


Pipette 10 µL of each ready-to use Standards, High Level Control, Low Level Control and Diluted Samples into the respective wells of microtiter plate.
A1: Standard A
B1: Standard B
C1: Standard C
D1: Standard D
E1: Standard E
F1: High Level Control
G1: Low Level Control
H1 and on: Sample (Serum / Plasma) A


Cover the plate with adhesive foil. Incubate 30 min at room temperature (18- 25°C).


Remove adhesive foil. Discard incubation solution. Wash plate 3 times each with 300µL of diluted. Wash Buffer. Remove excess solution by tapping the inverted plate on a paper towel.


Pipette 100 µL of ready-to use HRP-Conjugated Probe into each well.


Cover the plate with adhesive foil. Incubate 30 min at room temperature (18- 25°C).


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.


Pipette 100 µL of TMB Substrate Solution into each well.


Incubate 10 min (without adhesive foil) at room temperature (18-25°C) in the dark


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.


Measure optical density with a photometer at 450/650 nm within 30 min after pipetting of the Stop Solution.


Opdivo is a registered trademark of Bristol-Myers Squibb Company.