Etanercept ELISA Assay Kit

$955.00

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

Etanercept ELISA Assay Kit

The Etanercept ELISA Assay Kit is For Research Use Only

Size: 12×8 wells
Sensitivity: <2.5 ng/mL
Standard Range: 2.5-160 ng/ml
Incubation Time: 1.5 hours
Sample Type: Serum, Plasma, Cell Culture Supernatants
Sample Size: 100 µL
Alternative Name: Enbrel


Assay Principle

The method employs the quantitative sandwich enzyme immunoassay technique. Antibodies to Etanercept are pre-coated onto microwells. Samples and standards are pipetted into microwells and human Etanercept present in the sample are bound by the capture antibody. Then, a HRP (horseradish peroxidase) conjugated anti-Etanercept 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 Etanercept in the sample. Color development is then stopped by addition of stop solution. Absorbance is measured at 450 nm.


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.


Related Products to Etanercept ELISA

Anti-Etanercept (Enbrel) ELISA
Etanercept mAb-based ELISA Assay
Adalimumab (Humira) ELISA Kit

Additional Information

Assay Background


Etanercept is a biopharmaceutical that treats autoimmune diseases by interfering with tumor necrosis factor (TNF, a soluble inflammatory cytokine) by acting as a TNF inhibitor. It has U.S. F.D.A. approval to treat rheumatoid arthritis, juvenile rheumatoid arthritis and psoriatic arthritis, plaque psoriasis and ankylosing spondylitis. TNF-alpha is the “master regulator” of the inflammatory (immune) response in many organ systems.Autoimmune diseases are caused by an overactive immune response. Etanercept has the potential to treat these diseases by inhibiting TNF-alpha.

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 50 μl of Assay Diluent in each well.
3. Add 100 μl of Standards or Samples into the respective wells.
4. Cover the plate and incubate for 60 minutes at 37°C
5. 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.
6. Pipette without delay in the same order 100 μl of Anti-Etanercept: HRP Conjugate into each well.
7. Cover the plate and incubate for 60 minutes at 37°C
8. 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.
9. Add 100 μl of TMB Substrate in each well.
10. 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.
11. Pipette out 100 μl of Stop Solution. Wells should turn from blue to yellow in color.
12. Read the absorbance at 450 nm with a microplate reader.

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