PE Rat Anti-Mouse IL-3
Clone MP2-8F8 (RUO)
- Brand BD Pharmingen™
- Concentration 0.2 mg/ml
- Isotype Rat IgG1
- Reactivity Mouse (QC Testing)
Intracellular staining (flow cytometry) (Routinely Tested)
- Immunogen Recombinant mouse IL-3
- Storage Buffer Aqueous buffered solution containing ≤0.09% sodium azide.
- Regulatory Status RUO
Regulatory Status Legend
The MP2-8F8 antibody reacts with mouse interleukin-3 (IL-3). The immunogen used to generate the MP2-8F8 hybridoma was COS-expressed recombinant mouse IL-3.
- 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.
- 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.
Immunofluorescent Staining and Flow Cytometric Analysis: The MP2-8F8 antibody is useful for immunofluorescent staining and flow cytometric analysis to identify and enumerate IL-3 producing cells within mixed cell populations. PE-conjugated MP2-8F8 antibody (Cat. No. 554383) is especially suitable for these studies (see Figure, left panel). For optimal immunofluorescent staining with flow cytometric analysis, this anti-cytokine antibody should be titrated (≤ 0.25 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 conjugated MP2-8F8 antibody with a molar excess of ligand (e.g., recombinant mouse IL-3; Cat No. 554579) prior to staining, or 2) pre-block the fixed/permeabilized cells with unlabeled MP2-8F8 antibody (Cat. No. 554380, 554381) prior to staining. The staining technique and use of blocking controls are described in detail by C. Prussin and D. Metcalfe. A suitable rat IgG1 isotype control for assessing the level of background staining on paraformaldehyde-fixed/saponin-permeabilized mouse or human cells is PE-R3-34 (Cat. No. 554685); use at comparable concentrations to antibody of interest (e.g., ≤ 0.25 µg mAb/1 million cells).
Neutralization: The NA/LE™ MP2-8F8 antibody (Cat. No. 554379) is useful for neutralization of mouse IL-3 bioactivity.
ELISA Capture: The purified MP2-8F8 antibody (Cat. No. 554380) is useful as a capture antibody for a sandwich ELISA for measuring mouse IL-3 protein levels. For specific methodology, please visit the protocols section or chapter on ELISA in the Immune Function Handbook, both of which are posted on our web site, www.bdbiosciences.com. For testing mouse IL-3 in complex biological fluids such as serum or plasma, our mouse IL-3 specific OptEIA™ sandwich ELISA set is recommended (Cat. No. 555228).