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      PE Mouse Anti-Human Wiskott-Aldrich Syndrome Protein
      PE Mouse Anti-Human Wiskott-Aldrich Syndrome Protein
      Flow cytometric analysis of WASP expression in Human peripheral blood lymphocytes. Human peripheral blood mononuclear cells (PBMC) were fixed using BD Cytofix™ Fixation Buffer (Cat. No. 554655), washed and then permeabilized BD Perm/Wash™ Buffer (Cat. No. 554723).The cells were stained in BD Perm/Wash™ Buffer with either PE Mouse IgG2a, κ Isotype Control (Cat. No. 565363; dashed line histogram) or PE Mouse Anti-Human WASP antibody (Cat. No. 571364/571365; solid line histogram) at 0.5 µg/test. The  fluorescence histogram showing the expression of WASP (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ Flow Cytometer System and FlowJo™ Software. Data shown on this Technical Data Sheet are not lot specific.
      PE Mouse Anti-Human Wiskott-Aldrich Syndrome Protein
      Flow cytometric analysis of WASP expression in Human peripheral blood lymphocytes. Human peripheral blood mononuclear cells (PBMC) were fixed using BD Cytofix™ Fixation Buffer (Cat. No. 554655), washed and then permeabilized BD Perm/Wash™ Buffer (Cat. No. 554723).The cells were stained in BD Perm/Wash™ Buffer with either PE Mouse IgG2a, κ Isotype Control (Cat. No. 565363; dashed line histogram) or PE Mouse Anti-Human WASP antibody (Cat. No. 571364/571365; solid line histogram) at 0.5 µg/test. The  fluorescence histogram showing the expression of WASP (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ Flow Cytometer System and FlowJo™ Software. Data shown on this Technical Data Sheet are not lot specific.
      Product Details
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      BD Pharmingen™
      WASP
      Human (QC Testing), Mouse (Predicted)
      Mouse BALB/c IgG2a, κ
      Human WASP Recombinant Protein
      Intracellular staining (flow cytometry) (Routinely Tested)
      0.2 mg/ml
      Aqueous buffered solution containing ≤0.09% sodium azide.
      RUO


      Preparation And Storage

      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. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
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      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.

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      Product Notices

      1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
      2. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
      3. An isotype control should be used at the same concentration as the antibody of interest.
      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. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      6. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
      7. For U.S. patents that may apply, see bd.com/patents.
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      571365 Rev. 1
      Antibody Details
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      5A5

      Wiskott-Aldrich syndrome (WAS) is an X-linked recessive immunodeficiency disease caused by mutations in the gene encoding WAS protein (WASP).  The disease is characterized by a spectrum of clinical signs, including thrombocytopenia, eczema, susceptibility to opportunistic and pyogenic infections, and B-cell lymphomas associated with Epstein-Barr virus.  Furthermore, patients' blood cells display morphological abnormalities that can be associated with an impaired cytoskeleton.  WASP is a member of a family of highly conserved proteins that link signaling pathways to the actin cytoskeleton.  These members include WASP, N-WASP (neuronal), and SCAR/WAVE isoforms (Suppressor of cAMP Receptor/WASP family Verprolin-homologous protein) that share two main regions of homology: a proline-rich domain and a carboxyl terminal domain that binds to the Arp2/3 complex.  The Arp2/3 complex initiates actin filament assembly in motile cells and formation of the immunological synapse between activated T lymphocytes and antigen-presenting cells.  WASP is a central regulator of the actin cytoskeleton in hematopoietic cells that is itself regulated by multiple signaling pathways.

      The 5A5 antibody recognizes human WASP; it does not cross react with N-WASP.  It has been reported to detect WASP in lysates of hematopoietic cells and cell lines, except for neutrophils, from normal donors, but not from a group of patients having mutations of the WAS gene.

      571365 Rev. 1
      Format Details
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      PE
      R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is 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 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
      altImg
      PE
      Yellow-Green 488 nm, 532 nm, 561 nm
      496 nm, 566 nm
      576 nm
      571365 Rev.1
      Citations & References
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      View product citations for antibody "571365" on CiteAb

      Development References (10)

      1. Burns S, Cory GO, Vainchenker W, Thrasher AJ. Mechanisms of WASp-mediated hematologic and immunologic disease. Blood. 2004; 104(12):3454-3462. (Biology).
      2. Caron E. Regulation of Wiskott-Aldrich syndrome protein and related molecules. Curr Opin Cell Biol. 2002; 14:82-87. (Biology). View Reference
      3. Chiang SCC, Vergamini SM, Husami A, et al. Screening for Wiskott-Aldrich syndrome by flow cytometry.. J Allergy Clin Immunol. 2018; 142(1):333-335.e8. (Clone-specific: Flow cytometry). View Reference
      4. Descatoire M, Fritzen R, Rotman S, et al. Critical role of WASp in germinal center tolerance through regulation of B cell apoptosis and diversification.. Cell Rep. 2022; 38(10):110474. (Biology). View Reference
      5. Kawai S, Minegishi M, Ohashi Y, et al. Flow cytometric determination of intracytoplasmic Wiskott-Aldrich syndrome protein in peripheral blood lymphocyte subpopulations. J Immunol Methods. 2002; 260:195-205. (Immunogen: Flow cytometry, Immunoprecipitation, Western blot). View Reference
      6. Nakajima M, Yamada M, Yamaguchi K, et al. Possible application of flow cytometry for evaluation of the structure and functional status of WASP in peripheral blood mononuclear cells.. Eur J Haematol. 2009; 82(3):223-30. (Biology). View Reference
      7. Orange JS, Ramesh N, Remold-O'Donnell, et al. Wiskott-Aldrich syndrome protein is required for NK cell cytotoxicity and colocalizes with actin to NK cell-activatting immunologic synapses. Proc Natl Acad Sci U S A. 2002; 99(17):11351-11356. (Clone-specific: Immunofluorescence, Western blot).
      8. Yamada M, Ohtsu M, Kobayashi I, et al. Flow cytometric analysis of Wiskott-Aldrich syndrome (WAS) protein in lymphocytes from WAS patients and their familial carriers.. Blood. 1999; 93(2):756-7. (Clone-specific: Flow cytometry). View Reference
      9. Zeng R, Cannon JL, Abraham RT, et al. SLP-76 coordinates Nck-dependent Wiskott-Aldrich syndrome protein recruitment with Vav-1/Cdc42-dependent Wiskott-Aldrich syndrome protein activation at the T cell-APC contact site. J Immunol. 2003; 171:1360-1368. (Biology).
      10. de la Fuente MA, Sasahara Y, Calamito M, et al. WIP is a chaperone for Wiskott-Aldrich syndrome protein (WASP).. Proc Natl Acad Sci U S A. 2007; 104(3):926-31. (Clone-specific: Flow cytometry). View Reference
      View All (10) View Less
      571365 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.