FITC Rat Anti-Mouse CD249 (Ly-51)
Clone 6C3 (RUO)
- Brand BD Pharmingen™
- Alternative Name Bp-1/6C3; Ly51; Ly-51; Enpep; Glutamyl aminopeptidase; APA; EAP
- Concentration 0.5 mg/ml
- Isotype Rat F344, also known as Fischer, CDF IgG2a, κ
- Reactivity Mouse (QC Testing)
Flow cytometry (Routinely Tested)
- Immunogen C57L mouse Pre-B lymphhoma cell line L1-2
- Storage Buffer Aqueous buffered solution containing ≤0.09% sodium azide.
- Regulatory Status RUO
Regulatory Status Legend
The 6C3 monoclonal antibody specifically recognizes CD249 which is also known as, 6C3/BP-1, Enpep, or Ly-51. CD249 is a glycoprotein cell-surface differentiation antigen, which was originally identified on pre-B cell lymphomas (spontaneous and chemical- or retrovirus-transformed, in vitro and in vivo). CD249 is a homodimeric cell-surface glycoprotein with 140-kDa subunits that has been identified to possess aminopeptidase A (APA) activity. The same antigen is expressed at high levels on bone marrow stromal cell lines which support in vitro B lymphopoiesis, on thymic dendritic cells and cortical epithelial cells, and on a wide variety of mouse and rat tissues known to possess APA activity. Subsets of normal bone marrow pre-B and B lymphocytes express low levels of CD249 which is rapidly up-regulated on the pre-B cells in the presence of IL-7. A role for the CD249 molecule in the IL-7-driven proliferation of B cell precursors has been postulated. However, B-cell abnormalities were not detected in CD249/Ly-51-deficient mice. Mature B lymphocytes, thymocytes, peripheral T lymphocytes, erythroid cells, and myeloid cells (with the exception of thymic dendritic cells) do not express CD249. The 6C3 antibody can be used to identify cortical epithelium in frozen sections of thymuses from normal, SCID, and TCR-transgenic mice. It is possible that the low level of CD249 antigen detected, by flow cytometry, on some thymocytes may be passively adsorbed from adjacent epithelial cells during preparation of the cell suspensions.
FITC, fluorescein isothiocyanate, is a fluorochrome with a molecular weight of 389 Da. FITC is sensitive to pH changes and photobleaching. Due to nearly identical excitation and emission properties but different spillover characteristics, FITC and Alexa Fluor® 488 cannot be used simultaneously. FITC is relatively dim and should be reserved for highly expressed markers whenever possible.
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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 FITC under optimum conditions, and unreacted FITC was removed.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to www.bdbiosciences.com/pharmingen/protocols for technical protocols.