Invitrogen™ Human CD163 Uncoated ELISA Kit

Description

ELISA

The Human sCD163 Uncoated ELISA Kit contains pre-matched antibody pairs, plates and reagents for performing quantitative enzyme linked immunosorbent assays (ELISA) to detect and quantify protein levels of human sC163. Wash Buffer and Stop Solution are needed to complete the ELISA reaction and are sold separately. Principle of the method ELISAs are designed to measure the amount of the target bound between a matched antibody pair. A target-specific antibody is coated to the bottom of the wells of a microplate, which is an overnight process. Samples, standards, or controls are then added into these wells and bind to the immobilized (capture) antibody. A sandwich is formed by the addition of the second (detector) antibody, a substrate solution is added that reacts with the enzyme-antibody-target complex to produce measurable signal. The intensity of this signal is directly proportional to the concentration of target present in the original specimen.

CD163 (M130 antigen, Ber-Mac3, Ki-M8, SM4) is a 130 kDa membrane glycoprotein, a member of the scavenger receptor cysteine-rich superfamily, and a receptor for the hemoglobin-haptoglobin complex. CD163 protects tissues from free hemoglobin-mediated oxidative damage, and may play a role in the uptake and recycling of iron, via endocytosis of hemoglobin/haptoglobin and subsequent breakdown of heme. CD163 is expressed exclusively on the cell surface of human monocytes and macrophages that evolve predominantly in the late phase of inflammation. Specifically, CD163 is present on all circulating monocytes and most tissue macrophages except those found in the mantle zone and germinal centers of lymphoid follicles, interdigitating reticulum cells and Langerhan's cells. CD163 is present on all CD14 positive monocytes, most CD64 positive monocytes, and shows higher expression on CD16 positive monocytes. CD163 is upregulated on mononuclear phagocytes by IL-10, IL-6 and dexamethasone. Lipopolysaccharide (LPS) and phorbol myristate acetate (PMA) both induce shedding of CD163 from the cell surface into plasma or cell supernatant. CD163 binds hemoglobin/haptoglobin complexes in a calcium-dependent and pH-dependent manner, and exhibits a higher affinity for complexes of hemoglobin and multimeric haptoglobin of HP1F phenotype than for complexes of hemoglobin and dimeric haptoglobin of HP1S phenotype. Further, CD163 also induces a cascade of intracellular signals that involves tyrosine kinase-dependent calcium mobilization, inositol triphosphate production and secretion of IL6 and CSF1.

Specifications

• Accession Number: Cd163; CD163 antigen; CD163 molecule; CD163v2; CD163v3; hemoglobin scavenger receptor; M130; macrophage-associated antigen; MM130; putative CD163 antigen; SCARI1; Scavenger receptor cysteine-rich type 1 protein M130; sCD163; Soluble CD163; Soluble sCD163
• Assay Sensitivity: μg/mL
• Conjugate: HRP
• Product Type: ELISA
• Sample Type: Plasma, Serum, Supernatant
• For Use With (Equipment): Colorimetric Microplate Reader
• Gene ID (Entrez): 9332
• Gene Symbol: CD163
• Kit Contents: Capture Antibody: Pre-titrated, Purified antibody, Detection Antibody: Pre-titrated, Biotin-conjugated antibody, Standard: Recombinant protein for generating standard curve and calibrating samples, 10X Coating Buffer: Buffer for plating the Capture Antibody, 5X ELISA/ELISPOT Diluent: Buffer for blocking and diluting the Detection Antibody and Enzyme, Enzyme: Pre-titrated Streptavidin-HRP Concentrate (100X), Substrate: 1X TMB Solution, Certificate of Analysis: Lot-specific instructions for dilution of antibodies and standards, 96 Well Plate: High affinity 96-well plate (included with product Cat. Nos. ending in suffixes -22, -76, -86)
• Label or Dye: HRP
• Quantity: 2 x 96 Tests
• Regulatory Status: RUO
• Sample Volume: Plasma 50 μL, Serum 50 μL, Supernatant 50 μL
• Storage Requirements: 2°C to 8°C
• Target: CD163
• Test Time: 1 hrs 45 mins
• Total Assay Time: 24 hrs 30 mins