PE Rat Anti-Human IL-2
Clone MQ1-17H12 (RUO)
- Brand BD Pharmingen™
- Alternative Name IL2; Interleukin-2; T-cell growth factor; TCGF
- Vol. Per Test 20 µl
- Isotype Rat IgG2a, κ
- Reactivity Human (QC Testing) Rhesus, Cynomolgus, Baboon (Tested in Development)
Intracellular staining (flow cytometry) (Routinely Tested)
- Immunogen Human IL-2 Recombinant Protein
- Storage Buffer Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
- Regulatory Status RUO
Regulatory Status Legend
The MQ1-17H12 monoclonal antibody specifically binds to the multifunctional cytokine, human Interleukin-2 (IL-2). IL-2 is produced by activated T cells and has multiple functions that can affect the growth, proliferation, differentiation and survival of many different target cell types including T cells, B cells, NK cells, monocytes and macrophages. The immunogen used to generate the MQ1-17H12 hybridoma was purified recombinant human IL-2 protein. The MQ1-17H12 antibody reportedly neutralizes the biological activity of human IL-2.
- 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
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
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.
- 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).
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Species testing during development may have been performed with a different format of the same clone. Selected applications have been tested for cross-reactivity.
- Cy is a trademark of GE Healthcare.
- Please refer to www.bdbiosciences.com/pharmingen/protocols for technical protocols.
Immunofluorescent Staining and Flow Cytometric Analysis: The PE-conjugated MQ1-17H12 antibody (Cat. No. 559334/560902/554566) can be used for multicolor immunofluorescent staining and flow cytometric analysis to identify and enumerate IL-2-producing cells within mixed cell populations (see image). This 100 Test Size formulation of the PE-conjugated MQ1-17H12 antibody has been pre-titrated to assure effective intracellular detection of human IL-2 using 20 µl/1 × 10^6 cells.
A useful control for demonstrating specificity of staining is either of the following: 1) pre-block the conjugated MQ1-17H12 antibody with a molar excess of ligand (e.g., recombinant human IL-2; MQ1-17H12 antibody, Cat. No. 554563) prior to staining. A suitable rat IgG2a isotype control for assessing the level of background staining on paraformaldehyde-fixed/saponin-permeabilized human cells is also available in a 100 Test Size formulation PE-R35-95 (Cat. No. 559317). For specific methodology, please visit the protocols section on our web site, http://www.bdbiosciences.com/resources/index.jsp
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. BD Perm/Wash™ Buffer (Cat. No 554723) contains the
permeabilization agent saponin and is useful for this purpose as described in the USAGE section on the following page.
1. Resuspend 1 × 10^6 fixed and permeabilized cells in 20 µl of the pre-titered antibody solution and 30 µl of 1× BD Perm/Wash™ Buffer (Cat. No. 554723).
2. Incubate the cell suspension for 15 minutes (at RT or 4°C).
3. Wash twice in 100 µl of 1× BD Perm/Wash™ Buffer (Cat. No. 554723).