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APC-H7 Mouse Anti-Human CD52
APC-H7 Mouse Anti-Human CD52
Flow cytometric analysis of human CD52 expression on human peripheral blood lymphocytes. Whole blood was stained with either APC-H7 Mouse IgG3, κ Isotype Control (Cat. No. 563627; dashed line histogram) or APC-H7 Mouse Anti-Human CD52 antibody (Cat. No. 563611; solid line histogram). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
Flow cytometric analysis of human CD52 expression on human peripheral blood lymphocytes. Whole blood was stained with either APC-H7 Mouse IgG3, κ Isotype Control (Cat. No. 563627; dashed line histogram) or APC-H7 Mouse Anti-Human CD52 antibody (Cat. No. 563611; solid line histogram). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
Product Details
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BD Pharmingen™
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
AB_2738316
Aqueous buffered solution containing BSA, protein stabilizer, 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 APC-H7 under optimum conditions, and unconjugated antibody and APC-H7 were removed.

Product Notices

  1. 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).
  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. Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
  5. 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.
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. BD APC-H7 is a tandem conjugate and an analog of APC-Cy7 with the same spectral properties. It has decreased intensity but it is engineered for greater stability and less spillover in the APC channel and consequently offers better performance than APC-Cy7. It has an absorption maximum of approximately 650 nm. When excited by light from a red laser, the APC fluorochrome can transfer energy to the cyanine dye, which then emits at a longer wavelength. The resulting fluorescent emission maximum is approximately 767 nm. BD recommends that a 750-nm longpass filter be used along with a red-sensitive detector such as the Hamamatsu R3896 PMT. As with APC-Cy7 special filters are required when using APC-H7 in conjunction with APC. Note: Although our APC-H7 products demonstrate higher lot-to lot consistency than other APC tandem conjugate products, and every effort is made to minimize the lot-to-lot variation in residual emission from APC, it is strongly recommended that every lot be tested for differences in the amount of compensation required and that individual compensation controls are run for each APC-H7 conjugate.
  8. Although BD APC-H7 is engineered to minimize spillover to the APC channel and is more stable and less affected by light, temperature, and formaldehyde-based fixatives, compared to other APC-cyanine tandem dyes, it is still good practice to minimize as much as possible, any light, temperature and fixative exposure when working with all fluorescent conjugates.
  9. Cy is a trademark of GE Healthcare.
  10. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
563611 Rev. 2
Antibody Details
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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.

563611 Rev. 2
Format Details
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APC-H7
The BD Horizon™ APC-H7 dye is a part of the BD APC red family of dyes. This tandem fluorochrome is comprised of a Allophycocyanin (APC) donor that has excitation maxima (Ex Max) of 659 nm and an acceptor dye, H7, with an emission maximum (Em Max) at 782 nm. APC-H7, driven by BD innovation, is designed to be excited by the Red (627-640 nm) laser and detected using an optical filter centered near 780 nm (e.g., a 760/60 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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APC-H7
Red 627-640 nm
659 nm
782 nm
563611 Rev.2
Citations & References
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Development References (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). View Reference
  2. Zola H. Leukocyte and stromal cell molecules : the CD markers. Hoboken, N.J.: Wiley-Liss; 2007.
563611 Rev. 2

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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.