| Product Name | Eotaxin 2 antibody [61016.11] |
|---|---|
| Antibody Type | Primary Antibodies |
| Product description | Mouse monoclonal [61016.11] to Eotaxin 2Eotaxin 2, also named MPIF2, Ckbeta6, and SCYA24, is a member of the CC chemokine family, based on the presence of the CC motif and homology with other known CC chemokines. Eotaxin 2 cDNA encodes a 119 amino acid residue precursor protein with a 26 amino acid residue signal peptide that is cleaved to generate a mature protein predicted to contain 93 amino acid residues with an N glycosylation site. Mature human eotaxin 2 has a predicted molecular mass of approximately 10.6 kDa. Compared to other CC chemokines, eotaxin 2 exhibits 40 %, 42 %, and 39 % amino acid identity to MCP3, MIP1 alpha, and eotaxin, respectively. Human CC chemokine eotaxin 2 maps to chromosome 7q11.23. Both eotaxin and eotaxin 2 activate and attract eosinophils and basophils. A receptor for human eotaxin has been identified and found to be the third numbered receptor in the C-C chemokine subfamily of receptors (CCR3. On eosinophils, the effects of eotaxin 2 is inhibited by an CCR3 antibody and cross-desensitized by eotaxin and MCP4, suggesting that all three CC chemokines act through CCR3. Eotaxin 2 mRNA is weakly expressed in activated monocytes and T lymphocytes. Recombinant eotaxin 2 induces chemotaxis of eosinophils, basophils, and resting T lymphocytes but not monocytes and activated T lymphocytes. Eotaxin 2 inhibits colony formation in myleloid progenitor cells. |
| Immunogen | Purified recombinant human eotaxin 2, expressed in E. coli as immunogen. |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG1 |
| Clone Number | 61016.11 |
| Host Species | Mouse |
| Tested Applications | ELISANeutWB |
| ELISA: Use at a concentration of 2 μg/ml. Neut: Use at a concentration of 1 - 5 μg/ml. WB: Use at a concentration of 1 - 2 μg/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 μl of the cytokine-antibody solution (containing recombinant human eotaxin 2 at a final concentration of 0.1 μg/ml and antibody at concentrations from 0.01 to 100 μg/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 μg/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. Optimal dilutions/concentrations should be determined by the end user. | |
| Species Reactivity | Human |
| Concentration | 0.5mg/ml |
| Purification | Protein G |
| Gene Synonyms | Ckb-6, CCL24, 6369, MPIF2, SCYA24, 602495, Eotaxin 2, Ckb6, Eotaxin2, MPIF-2, O00175, Ckb 6, MPIF 2 |
|---|---|
| Target | Eotaxin 2 |
| Tissue Specificity | When used as a capture antibody in human eotaxin 2 sandwich ELISAs, this antibody exhibits less than 0.03 % cross-reactivity with recombinant human eotaxin, MIP1 alpha, and MCP3. |
| Cellular Localization | Secreted |
| Application Notes | ELISA: Use at a concentration of 2 μg/ml. Neut: Use at a concentration of 1 - 5 μg/ml. WB: Use at a concentration of 1 - 2 μg/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 μl of the cytokine-antibody solution (containing recombinant human eotaxin 2 at a final concentration of 0.1 μg/ml and antibody at concentrations from 0.01 to 100 μg/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 μg/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. Optimal dilutions/concentrations should be determined by the end user. |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | For short-term storage, store at 4° C. For long-term storage, aliquot and store at -20ºC or below. Avoid multiple freeze-thaw cycles. |
| Storage Buffer | Preservative: None Constituents: PBS |
