BD, 559332, BD Pharmingen™ PE Rat Anti-Human IL-5

Reactivity: Human (QC Testing)
Isotype: Rat IgG2a
Immunogen: Recombinant human IL-5
Application: Intracellular staining (flow cytometry) (Routinely Tested)
Vol. Per Test: 20 µl
RRID: AB_397229
Storage Buffer: Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
Regulatory Status: RUO
Preparation And Storage: Preparation And Storage 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. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
Recommended Assay Procedures: Recommended Assay Procedures Immunofluorescent Staining and Flow Cytometric Analysis: The JES1-39D10 antibody is useful for immunofluorescent staining and flow cytometric analysis to identify and enumerate human IL-5 producing cells within mixed cell populations. The PE-conjugated JES1-39D10 antibody is especially suitable for these experiments (see Figure). This 100 Test Size formulation of the PE-conjugated JES1-39D10 antibody has been pretitrated to assure effective intracellular detection of human IL-5 using 20 µl/1 x 10e6 cells. For specific methodology, please visit our web site, www.bdbiosciences.com, and go to the protocols section or the chapter on intracellular staining in the Immune Function Handbook. A useful control for demonstrating specificity of staining is either of the following: 1) pre-block the PE-JES1-39D10 antibody with excess ligand (e.g., human IL-5, Cat. No. 554606) prior to staining, or 2) pre-block paraformaldehyde-fixed/saponin-permeabilized cells with unlabeled JES1-39D10 antibody (e.g., Cat. No. 554488) 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 for assessing the level of background staining on paraformaldehyde-fixed/saponin-permeabilized mouse or human cells is also available in a 100 Test Size formulation PE-R35-95 (Cat. No. 559317). Important Note: This pretitered antibody solution does not contain a cell permeabilization agent. It is necessary to include a cell permeabilization agent when using the pre-titered antibody solution to stain fixed and permeabilized cells. Perm/Wash™ Buffer (Cat. No. 554723) contains the permeabilization agent saponin and is useful for this purpose as described in the Usage section. USAGE 1.   Resuspend 1 x 10e6 fixed and permeabilized cells in 20 µl of the pre-titered antibody solution and 30 µl of 1X Perm/Wash™ Buffer (Cat. No. 554723). 2.   Incubate the cell suspension for 15 minutes (at room temperature or 4°C). 3.   Wash twice in 100 µl of 1X Perm/Wash™ Buffer (Cat. No. 554723).
Product Notices: Product Notices This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test). Since applications vary, each investigator should titrate the reagent to obtain optimal results. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors. 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. Source of all serum proteins is from USDA inspected abattoirs located in the United States.