Application

Capture antibody in ELISA. Recommended coating concentration 5 µg/mL. See suggested protocol.

Optimal working dilutions must be determined by the end user.

Suggested ELISA Protocol For Ovine IL-6

1. Coat high capacity microtitre wells (e.g. Nunc maxisorp) with monoclonal anti-IL-6 antibody (Chemicon Catalog Number MAB1004) at a concentration of 5 µg/mL in 0.1 M carbonate buffer pH 9.6 overnight at 4°C.

2. Wash wells 3 x with PBS + 0.05% Tween 20 (PBST).

3. Add samples to wells. Incubate at room temperature in a humidified box for 1 hour. Wash 3 x with PBST.

4. Dilute rabbit anti-IL-6 (Chemicon Catalog Number AB1839) 1:500 in PBST and add 100 µL to wells. Incubate at room temperature for 30 minutes. Wash 3 x with PBST.

5. Dilute anti-rabbit IgG-HRP to appropriate level in PBST and add 100 µL to wells. Incubate 30 minutes at room temperature. Wash 3 x with PBST.

6. Add 100 µL of TMB chromagen to each well. Incubate for 5-10 minutes and stop reaction with 50 µL of 2M sulfuric acid.

Notes:

The anti-rabbit IgG HRP must be adsorbed against mouse immunoglobulin, and should be checked for non-reactivity with coated plates.

Non-specific binding may be reduced by addition of protein such as skim milk powder to the diluents.

Assay sensitivity may be altered by adjusting incubation times and reagent concentrations.

Research Sub CategoryCytokines & Cytokine Receptors

Research CategoryInflammation & Immunology

This Anti-Interleukin-6 Antibody, clone 4B6 is validated for use in ELISA for the detection of Interleukin-6.

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Immunogen

Recombinant ovine IL-6

Legal Information

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

Other Notes

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

Physical form

Liquid in PBS containing 0.1% sodium azide.

Format: Purified

Specificity

Reacts with ovine IL-6. No reactivity detected to recombinant ovine IL-1beta, IL-8, MCP or TNFalpha.

SPECIES REACTIVITY:

The antibody is likely to react with caprine (goat) IL-6.

Storage and Stability

Maintain at -20°C in undiluted aliquots for up to 12 months. Avoid repeated freeze/thaw cycles.