PE Rat Anti-Human IL-3
Clone BVD3-1F9 (RUO)
- Brand BD Pharmingen™
- Concentration 0.2 mg/ml
- Isotype Rat IgG1
- Reactivity Human (QC Testing)
Intracellular staining (flow cytometry) (Routinely Tested)
- Immunogen Recombinant Human IL-3
- Storage Buffer Aqueous buffered solution containing ≤0.09% sodium azide.
- Regulatory Status RUO
Regulatory Status Legend
The BVD3-1F9 antibody reacts with human interleukin-3 (IL-3). The immunogen used to generate the BVD3-1F9 hybridoma was recombinant human IL-3. This is a weakly neutralizing antibody.
- Format PE
- Excitation Source Blue 488 nm,Green 532 nm,Yellow/Green 561 nm
- Excitation Max 496 nm
- Emission Max 578 nm
R-phycoerythrin (PE) is an accessory photosynthetic pigment found in red algae. It exists in vitro as a 240-kDa protein with 23 phycoerythrobilin chromophores per molecule. This makes PE one of the brightest fluorochromes for flow cytometry applications, but its photobleaching properties make it unsuitable for fluorescence microscopy.
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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.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Please refer to www.bdbiosciences.com/pharmingen/protocols for technical protocols.
- 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.
Immunofluorescent Staining and Flow Cytometric Analysis: The BVD3-1F9 antibody is useful for immunofluorescent staining and flow cytometric analysis to identify and enumerate IL-3 producing cells within mixed cell populations. The PE-conjugated BVD3-1F9 antibody (Cat. No. 554676) is especially suitable for these studies. 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 suitable rat IgG1 isotype control for assessing the level of background staining on paraformaldehyde-fixed/saponin-permeabilized human cells is R3-34 (Cat. No. 554685); use at comparable concentrations to antibody of interest (e.g., ≤ 0.5 µg mAb/1 million cells). A useful control for demonstrating specificity of staining is either of the following: 1) pre-block the conjugated BVD3-1F9 antibody with ligand (e.g., recombinant human IL-3; Cat No. 554604) prior to staining, or 2) pre-block the fixed/permeabilized cells with unlabeled BVD3-1F9 antibody prior to staining. The staining technique and use of blocking controls are described in detail by C. Prussin and D. Metcalfe.
ELISA: The biotinylated BVD3-1F9 antibody (Cat. No. 554674) is useful as a detection antibody for a sandwich ELISA for measuring human IL-3 protein levels. Biotinylated BVD3-1F9 antibody can be paired with the purified BVD8-3G11 antibody (Cat. No. 554672) as the capture antibody, with recombinant human IL-3 protein (Cat. No. 554604) as the standard.