Mouse Naive/Memory T Cell Panel
- Brand BD Pharmingen™
- Reactivity Mouse (QC Testing)
Flow cytometry (Routinely Tested)
- Regulatory Status RUO
Regulatory Status Legend
Note: The panel is 50 test size. Extra material for each component for an additional 10 tests is provided for instrument set up purposes.
The antibodies are supplied in an aqueous buffered solution containing BSA, protein stabilizer, and ≤ 0.09% sodium azide.
Multicolor immunofluorescent staining followed by flow cytometric analysis and sorting has led to the phenotypic and functional characterization of multiple peripheral T cell subsets. Three major T cell subsets have been well characterized, i.e., naïve, memory and effector T cells. Naïve mouse T cells have a relatively homogenous cell surface phenotype. They express high levels of the peripheral lymph node homing receptor, CD62L (L-selectin), and low to intermediate levels of the adhesion molecule, CD44 (Pgp-1). Effector T cells coexpress variable to low levels of CD62L and high CD44 levels whereas memory T cells coexpress variable to high levels of CD62L and high levels of CD44. The Mouse Naive/Memory T Cell Panel contains fluorescent antibodies (each optimized at 5 μl per test) that are specific for the cell surface antigens: CD44, CD62L, CD4 and CD3. The panel was designed to standardize the multicolor staining and flow cytometric characterization of the three major CD4+ T cell subsets that arise as a consequence of development or clonal expansion and differentiation driven by antigenic stimulation (eg, in response to allergens, infectious disease or vaccination).
This fluorescent antibody panel was flexibly designed to permit the researcher's incorporation of FITC- or Alexa Fluor® 488-conjugated antibodies/probes or cells expressing green fluorescent proteins into flow cytometric analysis of mouse T cell subsets.
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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.
The antibody was conjugated with PerCP-Cy5.5 under optimum conditions, and unconjugated antibody and free PerCP-Cy5.5 were removed. Storage of PerCP-Cy5.5 conjugates in unoptimized diluent is not recommended and may result in loss of signal intensity.
The antibody was conjugated to APC under optimum conditions, and unconjugated antibody and free APC were removed.
The antibody was conjugated with APC-Cy7 under optimum conditions, and unconjugated antibody and free APC-Cy7 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).
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
- PerCP-Cy5.5–labelled antibodies can be used with FITC- and R-PE–labelled reagents in single-laser flow cytometers with no significant spectral overlap of PerCP-Cy5.5, FITC, and R-PE fluorescence.
- PerCP-Cy5.5 is optimized for use with a single argon ion laser emitting 488-nm light. Because of the broad absorption spectrum of the tandem fluorochrome, extra care must be taken when using dual-laser cytometers, which may directly excite both PerCP and Cy5.5™. We recommend the use of cross-beam compensation during data acquisition or software compensation during data analysis.
- This product is subject to proprietary rights of Amersham Biosciences Corp. and Carnegie Mellon University and made and sold under license from Amersham Biosciences Corp. This product is licensed for sale only for research. It is not licensed for any other use. If you require a commercial license to use this product and do not have one return this material, unopened to BD Biosciences, 10975 Torreyana Rd, San Diego, CA 92121 and any money paid for the material will be refunded.
- This APC-conjugated reagent can be used in any flow cytometer equipped with a dye, HeNe, or red diode laser.
- APC-Cy7 is a tandem fluorochrome composed of Allophycocyanin (APC), which is excited by laser lines between 595 and 647 nm and serves as an energy donor, coupled to the cyanine dye Cy7™, which acts as an energy acceptor and fluoresces at 780 nm. BD Biosciences Pharmingen has maximized the fluorochrome energy transfer in APC-Cy7, thus maximizing its fluorescence emission intensity, minimizing residual emission from APC, and minimizing required electronic compensation in multilaser-laser flow cytometry systems. Note: Although every effort is made to minimize the lot-to-lot variation in residual emission from APC, it is strongly recommended that every lot be tested for differences in the amount of compensation required and that individual compensation controls are run for each APC-Cy7 conjugate.
- APC-Cy7 tandem fluorochrome emission is collected in a detector for fluorescence wavelengths of 750 nm and higher.
- Warning: Some APC-Cy7 and PE-Cy7 conjugates show changes in their emission spectrum with prolonged exposure to formaldehyde. If you are unable to analyze fixed samples within four hours, we recommend that you use BD™ Stabilizing Fixative (Cat. No. 338036).
- Cy is a trademark of GE Healthcare.
- Please refer to www.bdbiosciences.com/pharmingen/protocols for technical protocols.