Adalimumab ELISA

Adalimumab ELISA Assay

$1,030.00

The Adalimumab ELISA Assay Kit is used as an analytical tool for quantitative determination of Adalimumab in serum, plasma and cell culture supernatant. The Adalimumab ELISA Assay Kit is for research use only and not to be used for diagnostic procedures.

SKU: KBI1015 Categories: , ,

Adalimumab ELISA Assay

The Adalimumab ELISA Assay is For Research Use Only

Size: 12×8 wells
Sensitivity: 4.8 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: Humira

Assay Background for Adalimumab Assay

Adalimumab is a recombinant human IgG1 monoclonal antibody specific for human tumor necrosis factor alpha (TNF-α). Adalimumab is produced by recombinant DNA technology in a mammalian cell expression system and is purified by a process that includes specific viral inactivation and removal steps. Adalimumab binds specifically to (TNF-α) and blocks its interaction with the p55 and p75 cell surface TNF receptors. TNF is a naturally occurring cytokine that is involved in normal inflammatory and immune responses. Elevated levels of TNF are found in the synovial fluid of rheumatoid arthritis, including juvenile idiopathic arthritis, psoriatic arthritis, and ankylosing spondylitis patients and play an important role in both the pathologic inflammation and the joint destruction that are hallmarks of these diseases. Increased TNF levels are also found in psoriasis (Ps) plaques.

Sample Preparation and Storage for Adalimumab ELISA:
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 samples can be stored at -20°C or -80⁰C. Avoid repeated freeze-thaw cycles.

Related Products

Anti-Adalimumab (Humira) ELISA
TNF-alpha Neutralization Rate Adalimumab ELISA Assay
Bevacuzimab (Avastin) Assay Kit

Additional Information

Assay Principle

The method employs the quantitative sandwich enzyme immunoassay technique. Antibodies to Adalimumab are pre-coated onto microwells. Samples and standards are pipetted into microwells and human Adalimumab present in the sample are bound by the capture antibody. Then, a HRP (horseradish peroxidase) conjugated anti-Adalimumab 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 Adalimumab 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 100 μl of Standards or Samples into the respective wells.
3. Cover the plate and incubate for 60 minutes at 37°C
4. 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.
5. Pipette without delay in the same order 100 μl of Anti-Adalimumab: HRP Conjugate into each well.
6. Cover the plate and incubate for 60 minutes at 37°C
7. 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.
8. Add 100 μl of TMB Substrate in each well.
9. 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.
10. Pipette out 100 μl of Stop Solution. Wells should turn from blue to yellow in color.
11. Read the absorbance at 450 nm with a microplate reader.

Documents

Product Documents

 

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.

Publications

Citations

Brempelis, K.J, Cowan, C.M, Kreuser, S.A., et al. Genetically engineered macrophages persist in solid tumors and locally deliver therapeutic proteins to activate immune responses. J Immunother Cancer (2020) 10.1136/jitc-2020-001356