Human MCP 1 ELISA Kit is a sandwich Enzyme-Linked Immunosorbent Assay (sELISA) designed for the in vitro quantitative determination of human MCP 1 in serum, plasma, and cell culture supernatant.
Assay Type
Sandwich (quantitative)
Principle of Assay
Human MCP 1 ELISA Kit (A334837) employs the sandwich enzyme immunoassay technique for the quantitative measurement of human MCP 1 in serum, plasma, and cell culture supernatant. An antibody specific for MCP 1 has been pre-coated onto a 96-well microtiter plate. The standards and test samples are added into the wells and the MCP 1 present in each sample is bound to the wells by the immobilized antibody. Following incubation, the wells are washed and then incubated with Biotinylated Anti-MCP 1 Antibody, which binds the captured MCP 1 present in each well. Following incubation, unbound biotinylated detection antibody is removed by washing, and an HRP-Streptavidin conjugate is added to the wells and the microtiter plate is incubated. Following incubation and washing, TMB substrate solution is then used to visualize the HRP enzymatic reaction by catalysis to produce a blue-coloured product that changes to yellow after addition of acidic stop solution. The density of yellow is proportional to the amount of MCP 1 captured in each well. The concentration of MCP 1 can then be calculated by reading the O.D. absorbance at 450nm in a microplate reader and referring to the standard curve.
Components
This ELISA Kit contains 10X 600 µL Capture Antibodies, 10X 600 µL Detector Antibodies, 2 Lyophilized Recombinant Protein Vials, 40 mL Assay Diluent #11, 10X 20 mL Wash Buffer, 12 mL TMB Solution, 12 mL Stop Solution, Pre-Coated 96-Well H-Microplate (12 × 8 well strips), and 2 Plate Seals.
A representative standard curve for the Human MCP-1 ELISA showing optical density (O.D.) at 450 nm plotted against MCP-1 concentrations (pg/mL) in Assay Diluent #11 on a logarithmic scale. The curve demonstrates a strong linear relationship across the tested concentration range, confirming the assay’s accuracy and sensitivity for MCP-1 quantification.
Human MCP-1 concentrations were determined in undiluted, 2×, 4×, and 8× diluted human serum and plasma samples. Serum samples exhibited higher MCP-1 concentrations compared to plasma, with consistent detection across all dilutions, indicating reliable assay performance and minimal matrix interference.
Human MCP-1 levels were quantified in undiluted and serially diluted (2×, 4×, 8×, and 16×) PBMC stimulated supernatants. The results show consistent MCP-1 concentrations across dilutions, indicating strong assay linearity and minimal dilutional effects within the tested range.
Human MCP-1 levels were assessed in undiluted and serially diluted (2×, 4×, 8×, 16×, and 32×) PBMC unstimulated supernatants. MCP-1 concentrations remained relatively consistent across dilutions, suggesting reliable assay performance and minimal interference from matrix effects in unstimulated samples.
Percentage recovery of Human MCP-1 was evaluated in serum (red) and cell culture media (blue) samples at various dilutions (25%-1%). Both sample types demonstrated recoveries within acceptable assay performance ranges (approximately 95-115%), indicating minimal matrix interference and consistent assay accuracy across dilution factors.
Percentage recovery of Human MCP-1 was assessed in plasma at serial dilutions ranging from 10% to 0.3%. The recovery values remained consistent (approximately 95-105%) across all dilutions, indicating high assay accuracy and minimal matrix interference in plasma samples.
