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BV421 Rat Anti-Mouse CD45RB
BV421 Rat Anti-Mouse CD45RB
Flow cytometric analysis of CD45RB expression on mouse thymocytes. Mouse thymocytes were stained with either BD Horizon™ BV421Rat Anti-Mouse CD45RB antibody (Cat. No. 562849, solid line histogram) or BD Horizon™ BV421 Rat IgG2a, κ Isotype Control (Cat. No. 562602, dashed line histogram). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of viable cells. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System. Immunohistofluorescent analysis of CD45RB expression by cells within C57BL/6 mouse spleen (right panel). A mouse spleen 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 Horizon™ BV421 Rat Anti-Mouse CD45RB antibody (Cat. No. 562849, pseudo-colored green) and Alexa Fluor® 647 Rat Anti-Mouse CD3 Molecular Complex antibody (Cat. No. 557869, pseudo-colored red). Images were captured on a standard epifluorescence microscope. Original magnification, 20x.
Flow cytometric analysis of CD45RB expression on mouse thymocytes. Mouse thymocytes were stained with either BD Horizon™ BV421Rat Anti-Mouse CD45RB antibody (Cat. No. 562849, solid line histogram) or BD Horizon™ BV421 Rat IgG2a, κ Isotype Control (Cat. No. 562602, dashed line histogram). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of viable cells. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System. Immunohistofluorescent analysis of CD45RB expression by cells within C57BL/6 mouse spleen (right panel). A mouse spleen 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 Horizon™ BV421 Rat Anti-Mouse CD45RB antibody (Cat. No. 562849, pseudo-colored green) and Alexa Fluor® 647 Rat Anti-Mouse CD3 Molecular Complex antibody (Cat. No. 557869, pseudo-colored red). Images were captured on a standard epifluorescence microscope. Original magnification, 20x.
Product Details
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BD Horizon™
Ptprc; CD45R; CD45; LCA; Leukocyte common antigen; Ly-5; Lyt-4
Mouse (QC Testing)
Rat IgG2a, κ
Cloned Mouse TH2 cell lines
Flow cytometry (Routinely Tested), Immunofluorescence (Tested During Development)
0.2 mg/ml
19264
AB_2737836
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 BD Horizon Brilliant™ dyes are used in a multicolor flow cytometry panel.  Fluorescent dye interactions may cause staining artifacts which may affect data interpretation.  The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. When BD Horizon Brilliant Stain Buffer is used in in the multicolor panel, it should also be used in the corresponding compensation controls for all dyes to achieve the most accurate compensation. For the most accurate compensation, compensation controls created with either cells or beads should be exposed to BD Horizon Brilliant Stain Buffer for the same length of time as the corresponding multicolor panel. 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

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  3. An isotype control should be used at the same concentration as the antibody of interest.
  4. 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.
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
  7. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  8. 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.
  9. 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.
  10. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
565993 Rev. 1
Antibody Details
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16A

The 16A monoclonal antibody specifically recognizes an exon B-dependent epitope of CD45 lycoprotein, which is found at high density on peripheral B cells, T cytotoxic/suppressor cells, a subset of T helper cells, and most thymocytes, and at low density on macrophages and dendritic cells. CD45RB expression appears to decrease as T lymphocytes progress from naive to memory cells. In addition, subpopulations of CD4+ T cells which express high and low levels of CD45RB have different cytokine secretion profiles and mediate distinct immunological functions. CD25+  D4+ regulatory T (Treg) lymphocytes which control intestinal inflammation and autoimmunity express low levels of  CD45RB.  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 (designated A, B, and C, respectively) as well as 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.

565993 Rev. 1
Format Details
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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.
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BV421
Violet 405 nm
407 nm
423 nm
565993 Rev.1
Citations & References
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Development References (3)

  1. Bottomly K, Luqman M, Greenbaum L. A monoclonal antibody to murine CD45R distinguishes CD4 T cell populations that produce different cytokines. Eur J Immunol. 1989; 19(4):617-623. (Immunogen: Cell separation, Flow cytometry, Fluorescence microscopy, Immunoprecipitation). View Reference
  2. Dianzani U, Luqman M, Rojo J. Molecular associations on the T cell surface correlate with immunological memory. Eur J Immunol. 1990; 20(10):2249-2257. (Clone-specific). View Reference
  3. Ernst DN, Weigle WO, Noonan DJ, McQuitty DN, Hobbs MV. The age-associated increase in IFN-γ synthesis by mouse CD8+ T cells correlates with shifts in the frequencies of cell subsets defined by membrane CD44, CD45RB, 3G11, and MEL-14 expression. J Immunol. 1993; 151(2):575-587. (Clone-specific: Flow cytometry). View Reference
565993 Rev. 1

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.