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Alexa Fluor™ 488 Mouse Anti-Human CD172a/b
Alexa Fluor™ 488 Mouse Anti-Human CD172a/b
Multiparameter flow cytometric analysis of CD172a/b expression on human peripheral blood leucocyte populations. Human whole blood was stained with either Alexa Fluor™ 488 Mouse IgG1, κ Isotype Control (Cat. No. 567121; Left Plot) or Alexa Fluor™ 488 Mouse Anti-Human CD172a/b antibody (Cat. No. 567973/568065; Right Plot). Erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). The bivariate pseudocolor density plot showing the correlated expression of CD172a/b [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 FACSymphony™ A5 Cell Analyzer System and FlowJo™ software.
Multiparameter flow cytometric analysis of CD172a/b expression on human peripheral blood leucocyte populations. Human whole blood was stained with either Alexa Fluor™ 488 Mouse IgG1, κ Isotype Control (Cat. No. 567121; Left Plot) or Alexa Fluor™ 488 Mouse Anti-Human CD172a/b antibody (Cat. No. 567973/568065; Right Plot). Erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). The bivariate pseudocolor density plot showing the correlated expression of CD172a/b [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 FACSymphony™ A5 Cell Analyzer System and FlowJo™ software.
Product Details
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BD Pharmingen™
SIRP alpha/beta1; SIRPα/SIRPβ1; Signal Regulatory Protein α/β1
Human (QC Testing)
Mouse BALB/c IgG1, κ
Human SIRP alpha extracellular domain Recombinant Protein
Flow cytometry (Routinely Tested)
5 µl
VII 70259
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 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. This will 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. Alexa Fluor® 488 fluorochrome emission is collected at the same instrument settings as for fluorescein isothiocyanate (FITC).
  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. 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).
  6. An isotype control should be used at the same concentration as the antibody of interest.
  7. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  8. This product is provided under an intellectual property license between Life Technologies Corporation and BD Businesses. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The buyer cannot sell or otherwise transfer (a) this product (b) its components or (c) materials made using this product or its components to a third party or otherwise use this product or its components or materials made using this product or its components for Commercial Purposes. Commercial Purposes means any activity by a party for consideration and may include, but is not limited to: (1) use of the product or its components in manufacturing; (2) use of the product or its components to provide a service, information, or data; (3) use of the product or its components for therapeutic, diagnostic or prophylactic purposes; or (4) resale of the product or its components, whether or not such product or its components are resold for use in research. For information on purchasing a license to this product for any other use, contact Life Technologies Corporation, Cell Analysis Business Unit Business Development, 29851 Willow Creek Road, Eugene, OR 97402, USA, Tel: (541) 465-8300. Fax: (541) 335-0504.
  9. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  10. Alexa Fluor™ is a trademark of Life Technologies Corporation.
568065 Rev. 1
Antibody Details
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SE5A5

The SE5A5 monoclonal antibody specifically binds to a common epitope on CD172a/SIRPα (90 kDa) and CD172b/SIRPβ1 (50 kDa). These transmembrane glycoproteins are members of the Signal Regulatory Protein (SIRP) family that, in turn, belongs to the Immunoglobulin superfamily. The SIRP family is comprised of two subgroups, SIRPα and SIRPβ that are distinguished by the presence (α) or absence (β) of a cytoplasmic domain containing two immunoreceptor tyrosine-based inhibition motifs (ITIM). CD172a/SIRPα is expressed on CD34+ stem/progenitor cells, cardiomyocytes, monocytes, macrophages, granulocytes, dendritic cells, and in the central nervous system. It binds to CD47 and is implicated in mediating inhibitory signals via the ITIM/SHP-2 association. CD172b/SIRPβ1 does not possess a cytoplasmic domain but instead the transmembrane domain contains a positively-charged residue that can interact with another transmembrane protein, DAP-12/KARAP. DAP-12 has two immunoreceptor tyrosine-based activation motifs (ITAM) within its cytoplasmic domain that are thought to link CD172b to cellular activation signaling. CD172b is expressed on myeloid cells, including peripheral blood monocytes and granulocytes. It is not expressed on CD34+ cells. CD172a and CD172b have complementary roles in signal regulation and may work together in tuning certain cellular responses to stimuli.

568065 Rev. 1
Format Details
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Alexa Fluor™ 488
Alexa Fluor™ 488 Dye is part of the BD blue family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 494-nm and an emission maximum (Em Max) at 517-nm. Alexa Fluor™ 488 is designed to be excited by the Blue laser (488 nm) and detected using an optical filter centered near 520-nm (e.g., a 530/30-nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 488
Blue 488 nm
494 nm
517 nm
568065 Rev.1
Citations & References
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View product citations for antibody "568065" on CiteAb

Development References (7)

  1. Bühring HJ, Simmons DL, Vernon-Wilson E. Review—CD172—SIRP; signal regulatory protein. 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; 2002:35.
  2. Dietrich J, Cella M, Seiffert M, Bühring HJ, Colonna M. Cutting edge: signal-regulatory protein beta 1 is a DAP12-associated activating receptor expressed in myeloid cells. J Immunol. 2000; 164(1):9-12. (Biology). View Reference
  3. Dubois NC, Craft AM, Sharma P, et al. SIRPA is a specific cell-surface marker for isolating cardiomyocytes derived from human pluripotent stem cells. Nat Biotechnol. 2011; 29:1011-1018. (Biology). View Reference
  4. Ghannadan M, Hauswirth AW, Schernthaner GH, et al. Detection of novel CD antigens on the surface of human mast cells and basophils. Int Arch Allergy Immunol. 2002; 127(4):299-307. (Biology: Flow cytometry). View Reference
  5. Seiffert M, Brossart P, Cant C, et al. Signal-regulatory protein alpha (SIRPalpha) but not SIRPbeta is involved in T-cell activation, binds to CD47 with high affinity, and is expressed on immature CD34(+)CD38(-) hematopoietic cells.. Blood. 2001; 97(9):2741-9. (Clone-specific: Immunoprecipitation, Inhibition). View Reference
  6. Seiffert M, Cant C, Chen Z, et al. Human signal-regulatory protein is expressed on normal, but not on subsets of leukemic myeloid cells and mediates cellular adhesion involving its counterreceptor CD47. Blood. 1999; 94(11):3633-3643. (Immunogen: Flow cytometry, Functional assay, Immunoprecipitation, Inhibition). View Reference
  7. Simmons DL, Vernon-Wilson E. Structure and function of the signal regulatory proteins (SIRPs). 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; 2002:35-38.
View All (7) View Less
568065 Rev. 1

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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.