Description
The MP1-22E9 monoclonal antibody specifically binds to mouse Granulocyte-Macrophage Colony Stimulating Factor (GM-CSF). The immunogen used to generate the MP1-22E9 hybridoma was yeast-expressed recombinant mouse GM-CSF. This is a neutralizing antibody.
Immunofluorescent Staining and Flow Cytometric Analysis: The MP1-22E9 antibody is useful for immunofluorescent staining and flow cytometric analysis to identify and enumerate GM-CSF producing cells within mixed cell populations. The PE-conjugated MP1-22E9 antibody (Cat. No. 554406) is especially suitable for these experiments (see Figure). For optimal immunofluorescent staining and flow cytometric analysis, this anti-cytokine antibody should be titrated (≤0.5 µg mAb/million cells). For specific methodology, please visit the protocols section or chapter on intracellular staining in the Immune Function Handbook, both of which are posted on our web site, www.bdbiosciences.com. A useful control for demonstrating specificity of staining is either of the following: 1) pre-block the PE-MP1-22E9 antibody with ligand (e.g., mouse GM-CSF, Cat. No. 554586) prior to staining, or 2) pre-block paraformaldehyde-fixed/saponin-permeabilized cells with unlabeled MP1-22E9 antibody (Cat. No. 554404) prior to staining. The intracellular staining technique and use of blocking controls are described in detail by C. Prussin and D. Metcalfe. A suitable rat IgG2a isotype control immunoglobulin for assessing the level of background staining on paraformaldehyde-fixed/saponin-permeabilized mouse cells is the PE-R35-95 antibody (Cat. No. 554689); use at comparable concentrations to antibody of interest ELISA Capture: The purified MP1-22E9 antibody (Cat. No. 554404) is useful as a capture antibody for a sandwich ELISA for measuring mouse GM-CSF protein levels. The purified MP1-22E9 antibody can be paired with the biotinylated MP1-31G6 antibody (Cat. No. 554407) as the detection antibody, with recombinant mouse GM-CSF (Cat. No. 554586) as the standard. Neutralization: The NA/LE™ MP1-22E9 antibody (Cat. No. 554403) is useful for neutralization of mouse GM-CSF bioactivity. A suitable NA/LE™ rat IgG2a isotype control to match the MP1-22E9 antibody is the R35-95 antibody, Cat. No. 554687.
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with R-PE under optimum conditions, and unconjugated antibody and free PE were removed.
1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
3. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
4. Use of these products to measure activation antigens expressed on mononuclear cell subsets for the purpose of monitoring immunoregulatory status can fall under one or more claims of the following patents: US Patent Nos. 5,445,939, 5,656,446, 5,843,689; European Patent No. 319,543; Canadian Patent No. 1,296,622; Australian Patent No. 615,880; and Japanese Patent No. 2,769,156.
5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
6. An isotype control should be used at the same concentration as the antibody of interest.
