V500 Mouse anti-Mouse CD45.2
Clone 104 (RUO)
- Brand BD Horizon™
- Alternative Name Ly-5.2; T200; LCA; Leukocyte common antigen; Ptprc
- Concentration 0.2 mg/ml
- Isotype Mouse SJL IgG2a, κ
- Reactivity Mouse (QC Testing)
Flow cytometry (Routinely Tested)
- Immunogen B10.S mouse thymocytes and splenocytes
- Entrez Gene ID 19264
- Storage Buffer Aqueous buffered solution containing protein stabilizer, glycerol and ≤0.09% sodium azide.
- Regulatory Status RUO
Regulatory Status Legend
The 104 monoclonal antibody recognizes the CD45 (Leukocyte Common Antigen) present on all leucocytes of most mouse strains (eg, A, AKR, BALB/c, CBA/Ca, CBA/J, C3H/He, C57BL, C57BR, C57L, C58, DBA/1, DBA/2, NZB, SWR, 129). This alloantigen was originally named Ly-5.1, and this was the designation at the time that the antibody was characterized. The designation was later changed from Ly-5.1 to Ly-5.2 to conform with the convention that the .2 alloantigen designations be assigned to the C57BL/6 strain. mAb 104 has been reported not to react with leucocytes of the mouse strains expressing the CD45.1 alloantigen (eg, RIII, SJL/J, STS/A, and DA). CD45 is a member of the Protein Tyrosine Phosphatase (PTP) family: its intracellular (COOH-terminal) region contains two PTP catalytic domains, and the extracellular region is highly variable due to alternative splicing of exons 4, 5, and 6 (designated A, B, and C, respectively), plus differing levels of glycosylation. The CD45 isoforms detected in the mouse are cell type-, maturation-, and activation state-specific. The CD45 isoforms play complex roles in T-cell and B-cell antigen receptor signal transduction. The 104 antibody has been reported to inhibit some responses of B cells, from mice expressing the CD45.2 alloantigen, to certain antigens and LPS. In addition, reduction of serum IgG levels and amelioration of autoimmune renal pathology were reported in mAb 104-treated systemic lupus erythematosus-prone mice.
The antibody is conjugated to BD Horizon™ V500, which has been developed for use in multicolor flow cytometry experiments and is
available exclusively from BD Biosciences. It is excited by the Violet laser with an Ex max of 415 nm and Em Max at 500 nm. BD Horizon V500 conjugates emit at a similar wavelength to Amcyan yet exhibit reduced spillover into the FITC channel. For more information on BD Horizon V500, visit bdbiosciences.com/colors.
When compensating dyes in this spectral range (such as Horizon™ V500 and AmCyan), the most accurate compensation can be obtained using single stained cellular controls. Due to spectral differences between cells and beads in this channel, using BD CompBeads can result in spillover errors for V500 and AmCyan reagents. Therefore, the use of BD CompBeads or BD CompBeads Plus to determine spillover values for these reagents is not recommended. Different V500 reagents (e.g. CD4 vs. CD45) can have slightly different fluorescence spillover therefore, it may also be necessary to use clone specific compensation controls when using these reagents.
BD Horizon™ V500 is a novel organic dye excited by the violet laser. This dye offers improved brightness over Pacific Orange™ and reduced spillover into the FITC channel when compared to AmCyan. BD Horizon V500 cannot be used simultaneously with AmCyan or Pacific Orange™.
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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 BD Horizon™ V500 under optimum conditions, and unreacted BD Horizon™ V500 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.
- BD Horizon V500 has a maximum absorption of 415 nm and maximum emission of 500 nm. Before staining with this reagent, please confirm that your flow cytometer is capable of exciting the fluorochrome and discriminating the resulting fluorescence.
- 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.
- Please refer to www.bdbiosciences.com/pharmingen/protocols for technical protocols.