PerCP-Cy™5.5 Mouse Anti-Human CD4
Clone L200 (RUO)
- Brand BD Pharmingen™
- Alternative Name L3T4 ; T-cell surface antigen T4/Leu-3; W3/25 ; CD4 antigen (p55)
- Vol. Per Test 20 µl
- Isotype Mouse BALB/c IgG1, κ
- Reactivity Rhesus, Cynomolgus, Baboon (QC Testing) Human (Tested in Development)
Flow cytometry (Routinely Tested)
- Immunogen Human HPB-ALL Cell Line
- Storage Buffer Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
- Regulatory Status RUO
Regulatory Status Legend
The L200 monoclonal antibody specifically binds to the human form of the 56 kDa transmembrane glycoprotein, CD4, which is present on the T-helper/inducer subset of normal human donor peripheral blood lymphocytes. The L200 antibody also cross-reacts with a subset of CD3-positive peripheral blood lymphocytes, but not monocytes, of both Rhesus and Cynomolgus Macaque monkeys. Cross-reactivity on both lymphocytes and monocytes (weak) from baboons is also observed. CD4 distribution on lymphocytes is similar for both human and monkey cells, with the majority of CD4-positive lymphocytes being CD8-negative and lacking reactivity with antibodies to B- or NK-cell markers.
PerCP-Cy™5.5 is a tandem conjugate that combines PerCP with a cyanine dye. PerCP-Cy5.5 is not subject to photobeaching like PerCP and can be used with stream-in-air flow cytometers. It has less Fc receptor–mediated nonspecific staining than PE-Cy5. Additionally, the PerCP-Cy5.5 tandem conjugate is not as susceptible to fixative or light instability compared to APC-Cy7 and PE-Cy7.
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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 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.
- 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.
- 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.
- 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 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.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.