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BD Horizon™ BV421 Mouse Anti-Human CD4
Clone RPA-T4 (RUO)
Flow cytometric analysis of CD4 expression on human peripheral blood lymphocytes. Human whole blood was stained at 4°C with the BD Horizon™ BV421 Mouse anti-Human CD4 antibody (Cat. No. 562424/562425; solid line histogram) or with a BD Horizon™ BV421 Mouse IgG1, κ Isotype Control (Cat. No. 562438; dashed line histogram). The erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). The fluorescence histograms were derived from events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometry was performed using a BD FACSCanto™ II Flow Cytometer System.
Immunohistofluorescent analysis of CD4 expression by cells within human tonsil. A human tonsil cryosection (5 µm) was fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655), blocked with 5% goat serum and 1% BSA diluted in 1x PBS, and stained with BD Pharmingen™ Purified Mouse Anti-Human CD8 antibody (Cat. No. 555364) followed by BD Horizon™ BV480 Goat Anti-Mouse Ig second step antibody (Cat. No. 564877, pseudo-colored green). Sections were thoroughly washed, then stained with BD Horizon™ BV421 Mouse Anti-Human CD4 antibody (Cat. No. 562424/562425, pseudo-colored red) and Alexa Fluor® 488 Mouse Anti-Human CD19 antibody (Cat. No. 557697, pseudo-colored blue). Images were captured on a standard epifluorescence microscope. Original magnification, 20x.
Immunohistofluorescent analysis of CD4 expression by cells within human tonsil. A human tonsil cryosection (5 µm) was fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655), blocked with 5% goat serum and 1% BSA diluted in 1x PBS, and stained with BD Pharmingen™ Purified Mouse Anti-Human CD8 antibody (Cat. No. 555364) followed by BD Horizon™ BV480 Goat Anti-Mouse Ig second step antibody (Cat. No. 564877, pseudo-colored green). Sections were thoroughly washed, then stained with BD Horizon™ BV421 Mouse Anti-Human CD4 antibody (Cat. No. 562424/562425, pseudo-colored red) and Alexa Fluor® 488 Mouse Anti-Human CD19 antibody (Cat. No. 557697, pseudo-colored blue). Images were captured on a standard epifluorescence microscope. Original magnification, 20x.
Flow cytometric analysis of CD4 expression on human peripheral blood lymphocytes. Human whole blood was stained at 4°C with the BD Horizon™ BV421 Mouse anti-Human CD4 antibody (Cat. No. 562424/562425; solid line histogram) or with a BD Horizon™ BV421 Mouse IgG1, κ Isotype Control (Cat. No. 562438; dashed line histogram). The erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). The fluorescence histograms were derived from events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometry was performed using a BD FACSCanto™ II Flow Cytometer System.
Immunohistofluorescent analysis of CD4 expression by cells within human tonsil. A human tonsil cryosection (5 µm) was fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655), blocked with 5% goat serum and 1% BSA diluted in 1x PBS, and stained with BD Pharmingen™ Purified Mouse Anti-Human CD8 antibody (Cat. No. 555364) followed by BD Horizon™ BV480 Goat Anti-Mouse Ig second step antibody (Cat. No. 564877, pseudo-colored green). Sections were thoroughly washed, then stained with BD Horizon™ BV421 Mouse Anti-Human CD4 antibody (Cat. No. 562424/562425, pseudo-colored red) and Alexa Fluor® 488 Mouse Anti-Human CD19 antibody (Cat. No. 557697, pseudo-colored blue). Images were captured on a standard epifluorescence microscope. Original magnification, 20x.
Immunohistofluorescent analysis of CD4 expression by cells within human tonsil. A human tonsil cryosection (5 µm) was fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655), blocked with 5% goat serum and 1% BSA diluted in 1x PBS, and stained with BD Pharmingen™ Purified Mouse Anti-Human CD8 antibody (Cat. No. 555364) followed by BD Horizon™ BV480 Goat Anti-Mouse Ig second step antibody (Cat. No. 564877, pseudo-colored green). Sections were thoroughly washed, then stained with BD Horizon™ BV421 Mouse Anti-Human CD4 antibody (Cat. No. 562424/562425, pseudo-colored red) and Alexa Fluor® 488 Mouse Anti-Human CD19 antibody (Cat. No. 557697, pseudo-colored blue). Images were captured on a standard epifluorescence microscope. Original magnification, 20x.
Product Details
BD Horizon™
L3T4; T-cell surface antigen T4/Leu-3; W3/25; CD4 antigen (p55)
Human (QC Testing)
Mouse IgG1, κ
PHA-stimulated Human Peripheral Blood Mononuclear Cells
Flow cytometry (Routinely Tested), Immunofluorescence (Reported)
5 µl
IV T114; VI 6T-079
AB_11154417
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
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).
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.
- Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
- 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.
- 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.
- 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.
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Stain Buffer (FBS) RUO
Size
500 ML
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 ML
Cat No.
554657
Brilliant Stain Buffer RUO
Size
100 Tests
Cat No.
563794
Lysing Buffer RUO
Size
100 ML
Cat No.
555899
Lysing Solution 10X Concentrate IVD
Size
100 ML
Cat No.
349202
Fixation Buffer RUO
Size
100 ML
Cat No.
554655
562424 Rev. 3
Antibody Details
RPA-T4
The RPA-T4 monoclonal antibody specifically binds to CD4, a 59 kDa single-chain transmembrane glycoprotein that is expressed on T-helper/inducer cell populations. CD4 is also expressed on thymocyte subsets and at lower levels on monocytes and macrophages. CD4 functions as a co-receptor in MHC class II-restricted antigen-induced T cell activation and as a receptor for human immunodeficiency viruses (HIV). This antibody binds to the D1 domain (CDR1 and CDR3 epitopes) of the CD4 antigen and reacts with approximately 80% of thymocytes and 45% of peripheral blood lymphocytes. RPA-T4 is capable of blocking HIV-1, gp120, and inhibits syncytium formation.
The antibody was conjugated to BD Horizon™ BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 407-nm and Em Max at 421-nm, BD Horizon™ BV421 can be excited by the violet laser and detected in the standard Pacific Blue™ filter set (eg, 450/50-nm filter). BD Horizon™ BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue™ conjugates.
562424 Rev. 3
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
562424 Rev.3
Citations & References
View product citations for antibody "562424" on CiteAb
Development References (3)
-
Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:1-1182.
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Piatier-Tonneau D. CD4 workshop panel report. In: Kishimoto T. Tadamitsu Kishimoto .. et al., ed. Leucocyte typing VI : white cell differentiation antigens : proceedings of the sixth international workshop and conference held in Kobe, Japan, 10-14 November 1996. New York: Garland Pub.; 1997:49-54.
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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.
562424 Rev. 3
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.