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BB700 Mouse Anti-Human CD117
BB700 Mouse Anti-Human CD117

Flow cytometric analysis of CD117 expressed on TF-1 cells. Cells from the human TF-1 (Human erythroleukemia, ATCC Cat. No. CRL-2003) cell line were stained with either BD Horizon™ BB700 Mouse IgG1, κ Isotype Control (Cat. No. 566404; dashed line histogram) or the BD Horizon BB700 Mouse Anti-Human CD117 antibody (Cat. No. 566548/566549; solid line histogram). A fluorescence histogram showing CD117 expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable cells. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.

Flow cytometric analysis of CD117 expressed on TF-1 cells. Cells from the human TF-1 (Human erythroleukemia, ATCC Cat. No. CRL-2003) cell line were stained with either BD Horizon™ BB700 Mouse IgG1, κ Isotype Control (Cat. No. 566404; dashed line histogram) or the BD Horizon BB700 Mouse Anti-Human CD117 antibody (Cat. No. 566548/566549; solid line histogram). A fluorescence histogram showing CD117 expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable cells. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.

Product Details
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BD Horizon™
KIT; c-Kit; SCFR; PBT; Mast/stem cell growth factor receptor
Human (QC Testing)
Mouse IgG1, κ
Acute myelocytic leukemia blasts
Flow cytometry (Routinely Tested)
5 µl
V C009
3815
AB_2744268
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 with BD Horizon™ BB700 under optimum conditions, and unconjugated antibody and free BD Horizon BB700 were removed.

Recommended Assay Procedures

For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions.  More information can be found in the Technical Data Sheet for the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).

When setting up compensation, it is recommended to compare spillover values obtained from cells and BD™ CompBeads to ensure that beads will provide sufficiently accurate spillover values.

For optimal results, it is recommended to perform two washes after staining with antibodies. Cells may be prepared, stained with antibodies and washed twice with wash buffer per established protocols for immunofluorescent staining, prior to acquisition on a flow cytometer. Performing fewer than the recommended wash steps may lead to increased spread of the negative population.

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. 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. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  5. The manufacture, use, sale, offer for sale, or import of this product is subject to one or more patents or pending applications. This product, and only in the amount purchased by buyer, may be used solely for buyer’s own internal research, in a manner consistent with the accompanying product literature. No other right to use, sell or otherwise transfer (a) this product, or (b) its components is hereby granted expressly, by implication or by estoppel. Diagnostic uses require a separate license.
  6. BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
  7. BD Horizon Brilliant Blue 700 is covered by one or more of the following US patents: 8,455,613 and 8,575,303.
  8. Cy is a trademark of GE Healthcare.
  9. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
566548 Rev. 1
Antibody Details
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YB5.B8

The YB5.B8 monoclonal antibody specifically binds to CD117, which is also known as c-Kit, or Stem cell factor Receptor (SCFR). CD117 is a ~145 kDa type I transmembrane glycoprotein with tyrosine kinase activity. CD117 is present on hematopoietic progenitor cell subsets, thymocytes, mast cells, hepatocytes and histiocytes. CD117 serves as a cytokine receptor for steel factor (SLF), also known as stem cell factor (SCF), or mast cell growth factor (MGF). The interaction of CD117 and SLF is crucial to hematopoiesis, mast cell differentiation, melanogenesis, and germ cell development. The ability of the YB5.B8 antibody to block the binding of c-Kit ligand is still controversial.

The antibody was conjugated to BD Horizon BB700, which is part of the BD Horizon Brilliant™ Blue family of dyes.   It is a polymer-based tandem dye developed exclusively by BD Biosciences.  With an excitation max of 485 nm and an emission max of 693 nm, BD Horizon BB700 can be excited by the 488 nm laser and detected in a standard PerCP-Cy™5.5 set (eg, 695/40-nm filter). This dye provides a much brighter alternative to PerCP-Cy5.5 with less cross laser excitation off the 405 nm and 355 nm lasers.

566548 Rev. 1
Format Details
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BB700
The BD Horizon Brilliant™ Blue 700 (BB700) Dye is part of the BD Horizon Brilliant™ Blue family of dyes. This tandem fluorochrome is comprised of a polymer-technology dye donor with an excitation maximum (Ex Max) of 476-nm and an acceptor dye with an emission maximum (Em Max) at 695-nm. Driven by BD innovation, BB700 is designed to be excited by the blue laser (488-nm) and detected using an optical filter centered near 695-nm (e.g., a 695/20-nm bandpass filter). The donor dye can be excited by the Violet (405 nm) laser and the acceptor dye can be excited by the red (627–640 nm) laser resulting in cross-laser excitation and fluorescence spillover. BB700 Reagents are significantly brighter than equivalent PerCP or PerCP-Cy5.5 reagents and are less sensitive to photobleaching. In addition, BB700 shows much less excitation by the violet (407-nm) laser resulting in less spillover. BB700 has minimal yellow green (562-nm) excitation and is ideal for instruments with both blue (488-nm) and yellow green (562-nm) lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
altImg
BB700
Blue 488 nm
476 nm
695 nm
566548 Rev.1
Citations & References
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Development References (6)

  1. Ashman LK, Buhring HJ, Aylett GW, Broudy VC, Muller C. Epitope mapping and functional studies with three monoclonal antibodies to the c-kit receptor tyrosine kinase, YB5.B8, 17F11, and SR-1. J Cell Physiol. 1994; 158(3):545-554. (Biology). View Reference
  2. Ashman LK, Cambareri AC, To LB, Levinsky RJ, Juttner CA. Expression of the YB5.B8 antigen (c-kit proto-oncogene product) in normal human bone marrow.. Blood. 1991; 78(1):30-7. (Clone-specific: Cell separation, Flow cytometry, Immunofluorescence). View Reference
  3. Lerner NB, Nocka KH, Cole SR, et al. Monoclonal antibody YB5.B8 identifies the human c-kit protein product. Blood. 1991; 77(9):1876-1883. (Immunogen: Blocking, Flow cytometry, Functional assay, Inhibition, Radioimmunoassay). View Reference
  4. 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.
  5. Wypych J, Bennett LG, Schwartz MG, et al. Soluble kit receptor in human serum. Blood. 1995; 85(1):66-73. (Biology). View Reference
  6. Yarden Y, Kuang WJ, Yang-Feng T, et al. Human proto-oncogene c-kit: a new cell surface receptor tyrosine kinase for an unidentified ligand. EMBO J. 1987; 6(11):3341-3351. (Biology). View Reference
View All (6) View Less
566548 Rev. 1

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