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PE-Cy™5 Rat Anti-Human CD49f
PE-Cy™5 Rat Anti-Human CD49f

Flow cytometric analysis of CD49f expression on human peripheral blood lymphocytes. Human whole blood was stained with either PE-Cy™5 Rat IgG2a, κ Isotype Control (Cat. No. 551066; dashed line histogram) or PE-Cy™5 Rat Anti-Human CD49f (Cat. No. 551129; solid line histogram). Erythrocytes were lysed with BD PharmLyse™ Lysing Buffer (Cat. No. 555899). Fluorescent histograms depicting CD49f (or Ig isotype) expression were derived from gated events with the side and forward light-scattering characteristics of viable lymphocytes.

Flow cytometric analysis of CD49f expression on human peripheral blood lymphocytes. Human whole blood was stained with either PE-Cy™5 Rat IgG2a, κ Isotype Control (Cat. No. 551066; dashed line histogram) or PE-Cy™5 Rat Anti-Human CD49f (Cat. No. 551129; solid line histogram). Erythrocytes were lysed with BD PharmLyse™ Lysing Buffer (Cat. No. 555899). Fluorescent histograms depicting CD49f (or Ig isotype) expression were derived from gated events with the side and forward light-scattering characteristics of viable lymphocytes.

Product Details
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BD Pharmingen™
ITGA6; IThe GoH3 monoclonal ntegrin alpha-6; Integrin α6 chain; VLA-6; ITA6
Human (QC Testing), Mouse, Pig, Dog (Tested in Development)
Rat SD, also known as Sprague-Dawley (outbred) IgG2a, κ
Mouse mammary tumor cells
Flow cytometry (Routinely Tested)
20 µl
IV P55
3655
AB_394062
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 PE-Cy5 (formerly known as BD Cy-Chrome™) under optimum conditions, and unconjugated antibody and free PE-Cy5 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 cell and CompBead to ensure that BD Comp beads are appropriate for your specific cellular application.

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. An isotype control should be used at the same concentration as the antibody of interest.
  3. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  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. 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.
  6. PE-Cy5 is optimized for use with a single argon ion laser emitting 488-nm light. Because of the broad absorption spectrum of the PE-Cy5 tandem fluorochrome, extra care must be taken when using dual-laser cytometers which may directly excite both PE and Cy5™.
  7. PE-Cy5 is a tandem fluorochrome composed of R-phycoerythrin (PE), which is excited by the 488 nm light of an Argon ion laser and serves as an energy donor, coupled to the cyanine dye Cy5, which acts as an energy acceptor and fluoresces at 670 nm. BD Biosciences Pharmingen has maximized the fluorochrome energy transfer in PE-Cy5, thus maximizing its fluorescence emission intensity, minimizing residual emission from PE, and minimizing lot-to-lot variation.
  8. PE-Cy5 tandem fluorochromes have been reported to bind some classes of human macrophages and granulocytes via Fc receptors, and PE has been reported to bind to mouse B lymphocytes via Fc receptors. Preincubation of mouse leukocytes with Mouse BD Fc Block™ purified anti-mouse CD16/CD32 mAb 2.4G2 can reduce the non-specific binding of PE-Cy5-conjugated reagents to mouse B cells. However, PE-Cy5 conjugated reagents should not be used to stain splenocytes of SJL, NOD, and MRL mice as B lymphocytes and/or other leukocytes have been reported to non-specifically stain regardless of the use of Mouse BD Fc Block™ (the CD72c complex has been implicated for PE-Cy5 binding in these strains). Reagents conjugated to PE, PerCP, PerCP-Cy5.5, APC, and APC-Cy7 tandem fluorochrome can be used on leukocytes from these mouse strains.
  9. Cy is a trademark of GE Healthcare.
  10. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  11. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  12. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
  13. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
551129 Rev. 8
Antibody Details
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GoH3

The GoH3 monoclonal antibody specifically binds to CD49f which is also known as integrin α6 chain. CD49f is a ~150 kDa type I transmembrane glycoprotein that belongs to the integrin alpha chain family of extracellular matrix and cell adhesion receptors. The integrin α6 subunit associates with the integrin β1 chain (CD29) to form VLA-6 and with the integrin β4 chain (CD104) to form the integrin α6β4 complex, also known as the laminin and kalinin receptor. CD49f is expressed mainly on T cells, monocytes, platelets, epithelial cells, endothelial cells, perineural cells, and trophoblasts of placenta. GoH3 recognizes an extracellular epitope of integrin α6 on human, mouse and bovine cells. GoH3 has been reported to block the binding of integrin α6 to laminin P1 and E8 fragments.

551129 Rev. 8
Format Details
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PE-Cy5
PE-Cy5 dye is a part of the BD PE family of dyes. This tandem fluorochrome is comprised of a R-Phycoerythrin (PE) donor that has excitation maxima (Ex Max) of 496 nm and 566 nm and an acceptor dye, Cy™5, with an emission maximum (Em Max) at 670-nm. PE designed to be excited by the Blue (488-nm), Green (532-nm) and yellow-green (561-nm) lasers and detected using an optical filter centered near 670-nm (e.g., a 670/20-nm bandpass filter). The donor dye can be excited by the Blue (488-nm), Green (532-nm) and yellow-green (561-nm) lasers and the acceptor dye can be excited by the Red (627-640-nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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PE-Cy5
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
670 nm
551129 Rev.8
Citations & References
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Development References (3)

  1. Aumailley M, Timpl R, Sonnenberg A. Antibody to integrin alpha 6 subunit specifically inhibits cell-binding to laminin fragment 8. Exp Cell Res. 1990; 188(1):55-60. (Biology). View Reference
  2. Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:1-1182.
  3. Sonnenberg A, Daams H, Van der Valk MA, Hilkens J, Hilgers J. Development of mouse mammary gland: identification of stages in differentiation of luminal and myoepithelial cells using monoclonal antibodies and polyvalent antiserum against keratin. J Histochem Cytochem. 1986; 34(8):1037-1046. (Immunogen). View Reference
551129 Rev. 8

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