Application

Inhibition Assays – Inhibition of biological activity of recombinant and natural TNF-α.
Detection of TNF-α in two-site immunoassay.

Immunohistochemistry detected TNF-α.

TNF-α Neutralization Assay

1. Plate 2 x 10⁴ murine L929B cells into individual wells of 96-well (flat bottomed) microtiter plate.

2. Incubate overnight at 37°C in humidified 5% CO₂ incubator.

3. The next day, prepare serial dilutions of anti-TNF (human) in medium containing 2 µg/mL of actinomycin D, for a final volume of 50 µL

4. Add 50 µL of TNF (10 ng/mL final concentration) to each antibody dilution and incubate for 1 hour at 37°C.

5. Add 100 µL of the antibody/antigen mixture to the pre-seeded L929B cells.

6. Incubate 24 hours at 37°C, 5% CO₂.

7. Score wells for cytotoxicity.

Optimal working dilutions must be determined by the end user.

Detect Tumor Necrosis Factor-α using this Anti-Tumor Necrosis Factor-α Antibody, clone 2C8 validated for use in ELISA, IH, NEUT.

Immunogen

Human recombinant tumor necrosis factor α.

Legal Information

CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany

Linkage

Replaces: 04-1114

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Physical form

Format: Purified

Specificity

No cross-reactivity observed with rat neuronal tissue.

Human tumor necrosis factor-a (TNF-a)

Storage and Stability

Maintain at 2-8°C in undiluted aliquots for up to one year.