Human/Mouse Total COX-2 DuoSet IC ELISA

Standard Curve

The Human/Mouse Total COX-2 DuoSet IC ELISA specifically recognizes total COX-2, as shown by Western blot.

Lysate prepared from CCD-1070Sk human foreskin fibroblasts treated with 5.0 ng/mL of recombinant human (rh) IL-1 beta (Catalog # 201-LB) for 16 hours was incubated in wells coated with Human/Mouse Total COX-2 Capture Antibody. Unbound material was removed by washing and bound material was solubilized in SDS gel sample buffer. The same lysate and the captured protein were electrophoresed, transferred to a PVDF membrane, and immunoblotted with an Anti-COX-2 Polyclonal Antibody (Catalog # AF4198). Only the band corresponding to human/mouse COX-2 was detected in captured material.

Quantification of induced COX-2.

Human PBMCs, THP-1 human acute monocytic leukemia cells, and RAW 264.7 mouse monocyte/macrophage cells were cultured in the absence or presence of 100 nM Phorbol 12-myristate 13-acetate (PMA) for 48 hours and 1.0 µg/mL of Lipopolysaccharide (LPS) for an additional 24 hours. Human CCD-1070Sk cells were cultured in the absence or presence of 5.0 ng/mL of rhIL-1 beta for 16 hours. The lysates were quantified with this DuoSet IC ELISA. The same lysates were immunoblotted (inset) with an anti-COX-2 polyclonal antibody. The DuoSet IC ELISA results correlate well with the relative amounts of human/mouse COX-2 detected by Western blot.