BD Horizon™ RB780 Mouse Anti-Human CD103 (Integrin αE)
Clone Ber-ACT8 (RUO)
Multiparameter flow cytometric analysis of CD103 (Integrin αE) expression on Human peripheral blood leucocytes. Human whole blood was stained with either BD Horizon™ RB780 Mouse IgG1, κ Isotype Control (Cat. No. 568532; Left Plot) or with BD Horizon™ RB780 Mouse Anti-Human CD103 (Integrin αE) antibody (Cat. No. 569331/569332; Right Plot). DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. Erythrocytes were lysed with BD FACS Lysing Solution (Cat. No. 349202). A bivariate pseudocolor density plot showing the correlated expression of CD103 (Integrin αE) [or Ig Isotype control staining] versus side-light scatter (SSC-A) signals was derived from gated events with the forward and side-light scatter characteristics of intact leucocyte populations. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.
Flow cytometric analysis of CD103 (Integrin αE) expression on activated Human peripheral blood lymphocytes. Human peripheral blood mononuclear cells (PBMC) were stimulated with Phytohemagglutinin (PHA; 3 days). The cells were then stained with either BD Horizon™ RB780 Mouse IgG1, κ Isotype Control (dashed line histogram) or BD Horizon™ RB780 Mouse Anti-Human CD103 (Integrin αE) antibody (solid line histogram). The fluorescence histogram showing the expressed levels of CD103 (Integrin αE) [or Ig Isotype control staining] was derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) lymphocytes.
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Multiparameter flow cytometric analysis of CD103 (Integrin αE) expression on Human peripheral blood leucocytes. Human whole blood was stained with either BD Horizon™ RB780 Mouse IgG1, κ Isotype Control (Cat. No. 568532; Left Plot) or with BD Horizon™ RB780 Mouse Anti-Human CD103 (Integrin αE) antibody (Cat. No. 569331/569332; Right Plot). DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. Erythrocytes were lysed with BD FACS Lysing Solution (Cat. No. 349202). A bivariate pseudocolor density plot showing the correlated expression of CD103 (Integrin αE) [or Ig Isotype control staining] versus side-light scatter (SSC-A) signals was derived from gated events with the forward and side-light scatter characteristics of intact leucocyte populations. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.
Flow cytometric analysis of CD103 (Integrin αE) expression on activated Human peripheral blood lymphocytes. Human peripheral blood mononuclear cells (PBMC) were stimulated with Phytohemagglutinin (PHA; 3 days). The cells were then stained with either BD Horizon™ RB780 Mouse IgG1, κ Isotype Control (dashed line histogram) or BD Horizon™ RB780 Mouse Anti-Human CD103 (Integrin αE) antibody (solid line histogram). The fluorescence histogram showing the expressed levels of CD103 (Integrin αE) [or Ig Isotype control staining] was derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) lymphocytes.
Product Details
BD Horizon™
ITGAE; Integrin alpha-E; ITAE; HML-1 antigen; Mucosal lymphocyte 1; HUMINAE
Human (QC Testing)
Mouse BALB/c IgG1, κ
Human MAPS16 T Cell Line
Flow cytometry (Routinely Tested)
5 µl/test
V A067, BP047, S256
3682
Aqueous buffered solution containing ≤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 to the dye under optimum conditions and unreacted dye was removed.
Recommended Assay Procedures
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.
Product Notices
- When using high concentrations of antibody, background binding of this dye to erythroid fragments produced by ammonium chloride-based lysis, such as with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899), has been observed when the antibody conjugate was present during the lysis procedure. This may cause nonspecific staining of target cells, such as leukocytes, which have bound the resulting erythroid fragments. This background can be mitigated by any of the following: titrating the antibody conjugate to a lower concentration, fixing samples with formaldehyde, or removing erythrocytes before staining (eg, gradient centrifugation or pre-lysis with wash). This background has not been observed when cells were lysed with BD FACS™ Lysing Solution (Cat. No. 349202) after staining.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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.
- 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.
- 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.
- Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
- Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
RB780 Mouse IgG1, κ Isotype Control RUO
Size
50 µg
Cat No.
568532
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569331 Rev. 3
Antibody Details
Ber-ACT8
The Ber-ACT8 monoclonal antibody specifically binds to the human mucosal lymphocyte antigen 1 (HML-1). This is a 175 kDa type I transmembrane glycoprotein, also known as integrin αE (ITGAE, Integrin alpha-E) and CD103. It is found on >90% of intestinal intraepithelial lymphocytes (iIEL) associated with integrin β7. CD103 is also expressed on lamina propria T lymphocytes in the intestine and on phytohemagglutinin-stimulated peripheral blood lymphocytes. It is rarely expressed on resting peripheral blood lymphocytes. It has been suggested that CD103 may have an accessory function for activation of iIEL. CD103 has been reported as a useful tool in hairy cell leukemia research.
569331 Rev. 3
Format Details
RB780
The BD Horizon RealBlue™ 780 (RB780) Dye is part of the BD family of blue dyes. It is a tandem fluorochrome with an excitation maximum (Ex Max) at 498-nm and an emission maximum (Em Max) at 781-nm. Driven by BD innovation, RB780 can be used on both spectral and conventional cytometers and is designed to be excited by the Blue laser (488-nm) with minimal excitation by the 561-nm Yellow-Green laser. For conventional instruments equipped with a Blue laser (488-nm), RB780 can be used as an alternative to PE-Cy7 and we recommend using an optical filter centered near 780-nm (eg, a 780/60-nm bandpass filter). For spectral instruments equipped with a Blue laser (488-nm), it can be used in conjunction with PE-Cy7. RB780 is on average brighter than PE-Cy7 and has minimal spillover into Yellow-Green detectors.
RB780
Blue 488 nm
498 nm
781 nm
569331 Rev.3
Citations & References
View product citations for antibody "569331" on CiteAb
Development References (7)
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DiGiuseppe JA, Borowitz MJ. Clinical utility of flow cytometry in the chronic lymphoid leukemias. Semin Immunol. 1998; 25(1):6-10. (Biology). View Reference
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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.
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Kruschwitz M, Fritzsche G, Schwarting R, et al. Ber-ACT8: new monoclonal antibody to the mucosa lymphocyte antigen. J Clin Pathol. 1991; 44(8):636-645. (Immunogen: Blocking, Flow cytometry, Immunoaffinity chromatography, Immunocytochemistry (cytospins), Immunohistochemistry, Immunoprecipitation). View Reference
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Matutes E, Meeus P, McLennan K, Catovsky D. The significance of minimal residual disease in hairy cell leukaemia treated with deoxycoformycin: a long-term follow-up study. Br J Haematol. 1997; 98(2):375-383. (Biology). View Reference
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Micklem KJ, Dong Y, Willis A, et al. HML-1 antigen on mucosa-associated T cells, activated cells, and hairy leukemic cells is a new integrin containing the beta 7 subunit. Am J Pathol. 1991; 139(6):1297-1301. (Clone-specific: Immunoprecipitation). View Reference
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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.
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Zola H. Leukocyte and stromal cell molecules : the CD markers. Hoboken, N.J.: Wiley-Liss; 2007.
569331 Rev. 3
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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.