Mouse monoclonal [61016.11] to Eotaxin 2

ELISA: Use at a concentration of 2 ?g/ml. Neut: Use at a concentration of 1 - 5 ?g/ml. WB: Use at a concentration of 1 - 2 ?g/ml. Predicted molecular weight: 10.6 kDa. The detection limit for human eotaxin 2 is approximately 1 ng/lane under non-reducing and reducing conditions. Eotaxin 2 has the ability to neutralizethe biological activity of recombinant human eotaxin 2. To measure this activity, recombinant human eotaxin 2 is incubated with various concentrations of the antibody for 30 minutes at room temperature in a 96 well microplate. Following this preincubation period, 35 ?l of the cytokine-antibody solution (containing recombinant human eotaxin 2 at a final concentration of 0.1 ?g/ml and antibody at concentrations from 0.01 to 100 ?g/ml) is transferred to the lower compartment of a 96 well chemotaxis chamber. The chemotaxis chamber is then assembled using a PVP-free polycarbonate filter (5 micron pore size) and 2 x 10^6 cells/ well (CCR-3 transfected Y3 cells) are added to the top chamber. After incubation for 3 hours at 37 °C in a 5 % CO2 humidified incubator, the chamber is disassembled and the cells that have migrated through to the lower chamber are transferred to a working plate and stained using MTT. Absorbance is read on a microplate reader. The exact concentration of antibody required to neutralize recombinant human eotaxin activity is dependent on the cytokine concentration, cell type, growth conditions, and the type of activity. The Neutralization Dose50 (ND50) for the antibody is defined as that concentration required to yield one-half maximal inhibition of the eotaxin 2 activity on a responsive cell line, when eotaxin 2 is present at a concentration just high enough to elicit a maximum response. The Neutralization Dose50 (ND50) for this antibody is approximately 1 to 5 ?g/ml in the presence of 100 ng/ml of recombinant human eotaxin 2, measuring the chemotaxis of rat myeloid Y3 cells transfected with the human CCR3 gene. Not tested in other applications. Optimal dilutions/concentrations should be determined by the end user.

Type: Primary
Antigen: MICAL1
Clonality: Monoclonal
Clone: 61016.11
Conjugation:
Epitope:
Host: Mouse
Isotype:
Reactivity: Human