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BD Horizon™ BV421 Mouse Anti-Human CD25
Clone M-A251 (RUO)
Flow cytometric analysis of CD25 expression on unstimulated and stimulated human peripheral blood lymphocytes.
Left Panel: Whole blood was stained with either BD Horizon™ BV421 Mouse IgG1, κ Isotype Control (Cat. No. 562438; dashed line histogram) or BD Horizon™ BV421 Mouse Anti-Human CD25 antibody (Cat. No. 562442/562443; solid line histogram). The erythrocytes were subsequently lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899).
Right Panel: Phytohemagglutinin-stimulated (3 days) human peripheral blood mononuclear cells were stained with either BD Horizon™ BV421 Mouse Anti-Human CD25 antibody (solid line histogram) or with BD Horizon™ BV421 Mouse IgG1, κ Isotype Control (dashed line histogram).
The fluorescence histograms showing CD25 expression (or Ig Isotype control staining) were derived from gated events with the forward and side light-scatter characteristics of viable lymphocytes (Left Panel) or lymphoblasts (Right Panel). Flow cytometric analysis was performed using a BD FACSCanto™ II Flow Cytometer System.
Flow cytometric analysis of CD25 expression on unstimulated and stimulated human peripheral blood lymphocytes.
Left Panel: Whole blood was stained with either BD Horizon™ BV421 Mouse IgG1, κ Isotype Control (Cat. No. 562438; dashed line histogram) or BD Horizon™ BV421 Mouse Anti-Human CD25 antibody (Cat. No. 562442/562443; solid line histogram). The erythrocytes were subsequently lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899).
Right Panel: Phytohemagglutinin-stimulated (3 days) human peripheral blood mononuclear cells were stained with either BD Horizon™ BV421 Mouse Anti-Human CD25 antibody (solid line histogram) or with BD Horizon™ BV421 Mouse IgG1, κ Isotype Control (dashed line histogram).
The fluorescence histograms showing CD25 expression (or Ig Isotype control staining) were derived from gated events with the forward and side light-scatter characteristics of viable lymphocytes (Left Panel) or lymphoblasts (Right Panel). Flow cytometric analysis was performed using a BD FACSCanto™ II Flow Cytometer System.
Flow cytometric analysis of CD25 expression on unstimulated and stimulated human peripheral blood lymphocytes.
Left Panel: Whole blood was stained with either BD Horizon™ BV421 Mouse IgG1, κ Isotype Control (Cat. No. 562438; dashed line histogram) or BD Horizon™ BV421 Mouse Anti-Human CD25 antibody (Cat. No. 562442/562443; solid line histogram). The erythrocytes were subsequently lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899).
Right Panel: Phytohemagglutinin-stimulated (3 days) human peripheral blood mononuclear cells were stained with either BD Horizon™ BV421 Mouse Anti-Human CD25 antibody (solid line histogram) or with BD Horizon™ BV421 Mouse IgG1, κ Isotype Control (dashed line histogram).
The fluorescence histograms showing CD25 expression (or Ig Isotype control staining) were derived from gated events with the forward and side light-scatter characteristics of viable lymphocytes (Left Panel) or lymphoblasts (Right Panel). Flow cytometric analysis was performed using a BD FACSCanto™ II Flow Cytometer System.
Flow cytometric analysis of CD25 expression on unstimulated and stimulated human peripheral blood lymphocytes.
Left Panel: Whole blood was stained with either BD Horizon™ BV421 Mouse IgG1, κ Isotype Control (Cat. No. 562438; dashed line histogram) or BD Horizon™ BV421 Mouse Anti-Human CD25 antibody (Cat. No. 562442/562443; solid line histogram). The erythrocytes were subsequently lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899).
Right Panel: Phytohemagglutinin-stimulated (3 days) human peripheral blood mononuclear cells were stained with either BD Horizon™ BV421 Mouse Anti-Human CD25 antibody (solid line histogram) or with BD Horizon™ BV421 Mouse IgG1, κ Isotype Control (dashed line histogram).
The fluorescence histograms showing CD25 expression (or Ig Isotype control staining) were derived from gated events with the forward and side light-scatter characteristics of viable lymphocytes (Left Panel) or lymphoblasts (Right Panel). Flow cytometric analysis was performed using a BD FACSCanto™ II Flow Cytometer System.
Product Details
BD Horizon™
IL-2R; IL2RA; IL-2Rα; TCGFR; TAC antigen; p55
Human (QC Testing), Rhesus, Cynomolgus, Baboon (Tested in Development)
Mouse BALB/c IgG1, κ
Phytohemagglutinin stimulated human lymphocytes
Flow cytometry (Routinely Tested)
5 µl
IV A053
AB_11154578
Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
RUO
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 BV421 under optimum conditions, and unconjugated antibody and free BD Horizon BV421 were removed.
Recommended Assay Procedures
BD™ CompBeads can be used as surrogates to assess fluorescence spillover (Compensation). When fluorochrome conjugated antibodies are bound to CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and CompBead to ensure that BD Comp beads are appropriate for your specific cellular application.
For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).
Product Notices
- 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.
- 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.
- Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
- BD Horizon Brilliant Violet 421 is covered by one or more of the following US patents: 8,158,444; 8,362,193; 8,575,303; 8,354,239.
- BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Stain Buffer (FBS) RUO
Size
500 ML
Cat No.
554656
BV421 Mouse Anti-Human CD25 RUO
Size
100 Tests
Cat No.
562442
BV421 Mouse IgG1, k Isotype Control RUO
Size
50 µg
Cat No.
562438
Lysing Buffer RUO
Size
100 ML
Cat No.
555899
562443 Rev. 4
Antibody Details
M-A251
The M-A251 monoclonal antibody specifically binds to the 55 kDa type I transmembrane glycoprotein known as low-affinity interleukin-2 receptor alpha chain subunit (IL-2Rα). CD25 is expressed on regulatory T cells, activated lymphocytes (T and B), and monocytes. It associates with the IL-2Rβ/CD122 and IL-2Rγ/CD132 receptor chains to form the high-affinity IL-2R complex. CD25 expression on T and B lymphocytes is upregulated by antigenic or mitogenic stimulation. Soluble CD25/IL-2Rα is produced as a consequence of lymphocyte stimulation and is found in biological fluids following inflammatory responses.
The antibody was conjugated to BD Horizon BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max near 407 nm and Em Max near 421 nm, BD Horizon BV421 can be excited by the violet laser (405 nm) and detected with a 450/50 nm filter. BD Horizon BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific BlueTM conjugates. Due to nearly identical excitation and emission properties but different spillover characteristics, BD Horizon BV421, Pacific Blue, and BD Horizon V450 cannot be used simultaneously.
562443 Rev. 4
Format Details
BV421
The BD Horizon Brilliant Violet™ 421 (BV421) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology based dye has an excitation maximum (Ex Max) of 407-nm and an emission maximum (Em Max) at 423-nm. Driven by BD innovation, BV421 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 420-nm (e.g., a 431/28-nm or 450/50-nm bandpass filter). BV421 is an ideal alternative for V450 as it is approximately ten times brighter with less spillover into the BV510/V500 detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
BV421
407 nm
423 nm
562443 Rev.4
Citations & References
View product citations for antibody "562443" on CiteAb
Development References (2)
-
Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:1-1182.
-
Schlossman SF. Stuart F. Schlossman .. et al., ed. Leucocyte typing V : white cell differentiation antigens : proceedings of the fifth international workshop and conference held in Boston, USA, 3-7 November, 1993. Oxford: Oxford University Press; 1995.
562443 Rev. 4
Please refer to Support Documents for Quality Certificates
Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
Please refer to Support Documents for Quality Certificates
Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.