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APC Mouse Anti-Human CD177
APC Mouse Anti-Human CD177
Multiparameter flow cytometric analysis of CD177 expression on human peripheral blood granulocytes. Whole blood was stained with either APC Mouse IgG1, κ Isotype Control (Cat. No. 554681; Left Plot) or APC Mouse Anti-Human CD177 antibody (Cat. No. 567970/568022; Right Plot). Erythrocytes were lysed with BD FACS Lysing Solution (Cat. No. 349202). A bivariate pseudocolor density plot showing the correlated expression of CD177 (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™ Cell Analyzer System and FlowJo™ software.
Multiparameter flow cytometric analysis of CD177 expression on human peripheral blood granulocytes. Whole blood was stained with either APC Mouse IgG1, κ Isotype Control (Cat. No. 554681; Left Plot) or APC Mouse Anti-Human CD177 antibody (Cat. No. 567970/568022; Right Plot). Erythrocytes were lysed with BD FACS Lysing Solution (Cat. No. 349202). A bivariate pseudocolor density plot showing the correlated expression of CD177 (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™ Cell Analyzer System and FlowJo™ software.
Product Details
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BD Pharmingen™
Human neutrophil alloantigen 2a; HNA-2a; NB1; PRV-1; PRV1
Human (QC Testing)
Mouse IgG1, κ
Human Granulocytes
Flow cytometry (Routinely Tested)
5 µl
VI M-M17, BP-609; VII 70510
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 to the dye under optimum conditions and unconjugated antibody and free dye were 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

  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. This APC-conjugated reagent can be used in any flow cytometer equipped with a dye, HeNe, or red diode laser.
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. An isotype control should be used at the same concentration as the antibody of interest.
  8. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
567970 Rev. 1
Antibody Details
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MEM-166

The MEM-166 monoclonal antibody specifically binds to CD177 which is also known as NB1 (Neutrophil-specific antigen), HNA-2a (Human neutrophil alloantigen 2a), and PRV1 (Polycythemia rubra vera 1). CD177 is a glycophosphatidylinositol (GPI)-anchored plasma membrane glycoprotein (56-64 kDa). CD177 is expressed on subpopulations of neutrophils, neutrophillic metamyelocytes and myelocytes from 89 to 97% of healthy individuals. It is also expressed on secondary granules. Its expression is upregulated on granulocytes stimulated with the chemotactic peptide f-met-leu-phe (fMLP). Although the function of CD177 has not been fully established, some reports suggest its possible role as receptor molecule involved in leucocyte migration.

567970 Rev. 1
Format Details
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APC
Allophycocyanin (APC), is part of the BD family of phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 651 nm and an emission maximum (Em Max) at 660 nm. APC is designed to be excited by the Red (627-640 nm) laser and detected using an optical filter centered near 660 nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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APC
Red 627-640 nm
651 nm
660 nm
567970 Rev.1
Citations & References
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Development References (7)

  1. Clement LT, Lehmeyer JE, Gartland GL.. Identification of neutrophil subpopulations with monoclonal antibodies. Blood. 1983 February; 61(2):326-332. (Biology). View Reference
  2. Kissel K, Santoso S, Hofmann C, Stroncek D, Bux J. Molecular basis of the neutrophil glycoprotein NB1 (CD177) involved in the pathogenesis of immune neutropenias and transfusion reactions. Eur J Immunol. 2001; 31(5):1301-1309. (Clone-specific: Western blot). View Reference
  3. Matsuo K, Lin A, Procter JL, Clement L, Stroncek D. Variations in the expression of granulocyte antigen NB1. Transfusion. 2000 June; 40(6):654-662. (Biology). View Reference
  4. Sachs UJ, Andrei-Selmer CL, Maniar A, et al. The neutrophil-specific antigen CD177 is a counter-receptor for platelet endothelial cell adhesion molecule-1 (CD31). J Biol Chem. 2007; 282(32):23603-23612. (Biology). View Reference
  5. Stroncek D, Kissel K, Bux J. CD177: Neutrophil-specific antigen NB1. In: Mason D. David Mason .. et al., ed. Leucocyte typing VII : white cell differentiation antigens : proceedings of the Seventh International Workshop and Conference held in Harrogate, United Kingdom. Oxford: Oxford University Press; 2002:374-376.
  6. Stroncek DF, Shankar R, Litz C, Clement L. The expression of the NB1 antigen on myeloid precursors and neutrophils from children and umbilical cords. Transfus Med. 1998 June; 8(2):119-123. (Biology). View Reference
  7. Stroncek DF, Shankar RA, Noren PA, Herr GP, Clement LT. Analysis of the expression of NB1 antigen using two monoclonal antibodies. Transfusion. 1996 February; 36(2):168-174. (Biology). View Reference
View All (7) View Less
567970 Rev. 1

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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.