BD Pharmingen™ PE Rat Anti-Human IL-13

Immunofluorescent Staining and Flow Cytometric Analysis: The PE Rat Anti-Human IL-13 (Cat. No. 554571) can be used for multicolor immunofluorescent staining and flow cytometric analysis to identify and enumerate IL-13 producing cells within mixed cell populations (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 our website, https://www.bdbiosciences.com/en-us/resources/protocols.

A useful control for demonstrating specificity of staining is either of the following: 1) pre-block the PE Rat Anti-Human IL-13 antibody with ligand (e.g., human IL-13) prior to staining, or 2) pre-block paraformaldehyde-fixed/saponin-permeabilized cells with unlabeled Rat Anti-Human IL-13 antibody (e.g., Cat. No. 554570) 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 IgG1 isotype control immunoglobulin for assessing the level of background staining on paraformaldehyde-fixed/saponin-permeabilized mouse or human cells is the PE Rat IgG1, κ Isotype Control (Cat. No. 554685); use at comparable concentrations to antibody of interest (e.g., ≤ 0.5 µg mAb/1 million cells).

BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation).  When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells.   However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls.  It is strongly recommended that when using a reagent for the first time, users compare the spillover on cell and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.

1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
2. An isotype control should be used at the same concentration as the antibody of interest.
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. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
5. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
6. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).