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BV421 Mouse Anti-Human CD52
BV421 Mouse Anti-Human CD52

Multiparameter flow cytometric analysis of CD52 expression on human peripheral leucocyte populations. Human whole blood was stained with either BD Horizon™ BV421 Mouse IgG3, κ Isotype Control (Cat. No. 563314; Left Plot) or BD Horizon™ BV421 Mouse Anti-Human CD52 (Cat. No. 567789/567790; Right Plot). Erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). Bivariate pseudocolor density plots showing CD52 expression (or Ig Isotype control staining) versus side-light scatter (SSC-A) signals were derived from gated events with the side and forward light-scattering characteristics of intact leucocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.

Multiparameter flow cytometric analysis of CD52 expression on human peripheral leucocyte populations. Human whole blood was stained with either BD Horizon™ BV421 Mouse IgG3, κ Isotype Control (Cat. No. 563314; Left Plot) or BD Horizon™ BV421 Mouse Anti-Human CD52 (Cat. No. 567789/567790; Right Plot). Erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). Bivariate pseudocolor density plots showing CD52 expression (or Ig Isotype control staining) versus side-light scatter (SSC-A) signals were derived from gated events with the side and forward light-scattering characteristics of intact leucocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.

製品詳細
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BD Horizon™
Cambridge pathology 1 Ag; CAMPATH-1; Epididymal secretory protein E5; HE5
Human (QC Testing)
Mouse BALB/c IgG3, κ
Human T Lymphocytes
Flow cytometry (Routinely Tested)
5 µl
Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
RUO


調製と保管

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 to the dye under optimum conditions and unconjugated antibody and free dye were removed.

推奨アッセイ手順

BD® CompBeads can be used as surrogates to assess fluorescence spillover (Compensation). When fluorochrome conjugated antibodies are bound to BD® 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 BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.

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).

製品通知

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  3. 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.
  4. 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).
  5. An isotype control should be used at the same concentration as the antibody of interest.
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. 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.
  8. 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.
  9. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  10. Pacific Blue™ is a trademark of Life Technologies Corporation.
567789 Rev. 1
抗体の詳細
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4C8

The 4C8 monoclonal antibody specifically binds to CD52 which is also known as Cambridge pathology 1 antigen (CAMPATH-1) or Human epididymis-specific protein 5 (HE5). CD52 is a highly N-glycosylated, 25-29 kDa protein whose C-terminus is glycophosphatidylinositol anchored in the membrane. It is highly expressed on the surface of thymocytes and mature lymphocytes but not on their stem cell precursors. It is also expressed on monocytes, dendritic cells, eosinophils and epithelial cells of the epididymis and seminal vesicles but not on neutrophils, plasma cells, platelets or erythrocytes. Although its functional role is not well characterized, the CD52 antigen serves as an exquisitely sensitive target antigen for antibody and complement-mediated lysis of CD52-positive cells. Anti-CD52 antibodies are being used clinically to remove lymphocytes from transplanted bone marrow cell preparations and in the treatment of some malignant diseases.

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 Blue™ 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.

567789 Rev. 1
フォーマットの詳細
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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
567789 Rev.1
引用&参考文献
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開発者向け参考資料 (2)

  1. Masuyama J, Yoshio T, Suzuki K, et al. Characterization of the 4C8 antigen involved in transendothelial migration of CD26(hi) T cells after tight adhesion to human umbilical vein endothelial cell monolayers.. J Exp Med. 1999; 189(6):979-990. (Immunogen: Blocking, Flow cytometry, Stimulation, Western blot). 参考文献を見る
  2. Zola H. Leukocyte and stromal cell molecules : the CD markers. Hoboken, N.J.: Wiley-Liss; 2007.
567789 Rev. 1

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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.