Anti-Interleukin-8 Antibody, clone 8M6

Recombinant ovine IL-8

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

Flow cytometry: use 1 μg/test

Neutralizes recombinant ovine IL-8.

Western blotting

Optimal working dilutions must be determined by the end user.

Suggested ELISA Protocol For Ovine IL-8

1. Coat high capacity microtitre wells (e.g. Nunc maxisorp) with monoclonal anti-IL-8 antibody (Chemicon Catalog Number MAB1044) at a concentration of 5 μg/mL in 0.1 M carbonate buffer pH 9.6 overnight at 4ºC.

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-8 (Chemicon Catalog Number AB1840) 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 Category
Inflammation & Immunology

Research Sub Category
Cytokines & Cytokine Receptors

This Anti-Interleukin-8 Antibody, clone 8M6 is validated for use in ELISA, FC, NEUT, WB for the detection of Interleukin-8.

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

Format: Purified

Liquid in PBS containing 0.1% sodium azide.

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

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

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