BD Pharmingen™ Purified Rat Anti-Human IL-13

ELISA Capture: The purified JES10-5A2 (Cat. No. 554570) is useful as a capture antibody for a sandwich ELISA for specifically measuring human IL-13 protein levels. Purified JES10-5A2 antibody can be paired with the biotinylated clone B69-2 mouse IgG1 anti-human IL-13 detection antibody (Cat. No. 555054), with recombinant human IL-13 as the standard. Purified JES10-5A2 antibody should be titrated 1.0 - 4.0 µg/ml to determine the optimal concentration for ELISA capture. To obtain linear standard curves, doubling dilutions of human IL-13 protein ranging from ~2,000 to 15 pg/ml are recommended for inclusion in each ELISA plate. For specific methodology, please visit our website, www.bdbiosciences.com , and go to the protocols section or the chapter on ELISA in the Immune Function Handbook.

Note 1: This ELISA pair shows no cross-reactivity with any of the cytokines tested (e.g., mouse IL-1β, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-9, IL-10, IL-12 p70, IL-15, GM-CSF, IFN-γ, MCP-1, TCA-3, TNF; human IL-1α, IL-1β, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12 p70, IL-12 p40, IL-15, G-CSF, GM-CSF, IFN-γ, lymphotactin, MCP-1, MCP-2, MIP-1α, MIP-1β, NT-3, PDGF-AA, sCD23, SCF, TNF, LT-α, VEGF; rat IL-2, IL-4, IL-6, IL-10, GM-CSF, IFN-γ, TNF).

Note 2: This ELISA pair is recommended primarily for measuring cytokine from experimental cell culture systems. These ELISA reagents are not recommended for assay of serum or plasma samples.

Western Blot: The JES10-5A2 antibody (Cat. No. 554570) has been reported to be useful for Western blotting. Please note that this application is not routinely tested at BD Biosciences Pharmingen.

Neutralization: The NA/LE™ JES10-5A2 antibody (Cat. No. 554568) is useful for neutralization of human IL-13 bioactivity. A suitable NA/LE™ isotype control to match the JES10-5A2 antibody is the R3-34 antibody, (Cat. No. 554682).

IF/Flow: The JES10-5A2 antibody is useful for immunofluorescent staining and flow cytometric analysis to identify and enumerate IL-13 producing cells within mixed cell populations. PE-conjugated JES10-5A2 antibody (Cat. No. 554571) is especially suitable for these studies. For specific methodology, please visit our web site, w ww.bdbiosciences.com , and go to the protocols section or the chapter on intracellular staining in the Immune Function Handbook.

Immunohistochemistry: The JES10-5A2 antibody is also reported to be useful for immunohistochemical staining studies. The purified JES10-5A2 antibody (Cat. No. 554570) may be used for the immunofluorescent staining of human IL-13, when used in conjunction with an appropriate FITC-conjugated polyclonal anti-rat Ig with no reactivity towards human Igs. For immunoenzymatic staining, the purified JES10-5A2 antibody should be detected with a biotin conjugated polyclonal anti-rat Ig, followed by an enzyme-conjugated avidin or streptavidin. The paraformaldehyde/saponin method described by J. Andersson et al. is recommended for this staining. A recommended titration range of the antibody for this staining is 2-5 µg/ml, with the optimal concentration dependent upon various factors including the type of tissue stained and the secondary reagents used. A suitable rat IgG1 isotype control for assessing the level of background staining on paraformaldehyde-fixed/saponin-permeabilized human cells or tissue is purified R3-34 antibody (Cat. No. 559072); use at comparable concentrations to antibody of interest.

Note: this antibody is not routinely tested in the IHC application.

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.