Flow cytometric analysis of FLK-1 expression on bEnd.3 cells. bEnd.3 cells were stained either with an APC-Cy7 Rat IgG2a, κ Isotype Control (Cat. No. 552770; dashed line histogram) or with the APC-Cy7 Rat Anti-Mouse FLK-1 antibody (Cat. No. 561252; solid line histogram). The flow cytometric histograms were derived from events with the forward and side light-scatter characteristics of viable bEnd.3 cells. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.
Flow cytometric analysis of FLK-1 expression on bEnd.3 cells. bEnd.3 cells were stained either with an APC-Cy7 Rat IgG2a, κ Isotype Control (Cat. No. 552770; dashed line histogram) or with the APC-Cy7 Rat Anti-Mouse FLK-1 antibody (Cat. No. 561252; solid line histogram). The flow cytometric histograms were derived from events with the forward and side light-scatter characteristics of viable bEnd.3 cells. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.
Product Details
BD Pharmingen™
CD309; Fetal liver kinase 1; Kdr; VEGF Receptor-2; VEGFR-2; AVAS12;
Mouse (QC Testing)
Rat WI, also known as Wistar (outbred) IgG2a, κ
Mouse Flk-1 Recombinant Protein
Flow cytometry (Routinely Tested)
0.2 mg/ml
AB_10643607
Aqueous buffered solution containing protein stabilizer 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 with APC-Cy7 under optimum conditions, and unconjugated antibody and free APC-Cy7 were removed.
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- APC-Cy7 is a tandem fluorochrome composed of Allophycocyanin (APC), which is excited by laser lines between 595 and 647 nm and serves as an energy donor, coupled to the cyanine dye Cy7™, which acts as an energy acceptor and fluoresces at 780 nm. BD Biosciences Pharmingen has maximized the fluorochrome energy transfer in APC-Cy7, thus maximizing its fluorescence emission intensity, minimizing residual emission from APC, and minimizing required electronic compensation in multilaser-laser flow cytometry systems. Note: Although every effort is made to minimize the lot-to-lot variation in residual emission from APC, it is strongly recommended that every lot be tested for differences in the amount of compensation required and that individual compensation controls are run for each APC-Cy7 conjugate.
- APC-Cy7 tandem fluorochrome emission is collected in a detector for fluorescence wavelengths of 750 nm and higher.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Warning: Some APC-Cy7 and PE-Cy7 conjugates show changes in their emission spectrum with prolonged exposure to formaldehyde. If you are unable to analyze fixed samples within four hours, we recommend that you use BD™ Stabilizing Fixative (Cat. No. 338036).
- Cy is a trademark of GE Healthcare.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
APC-Cy™7 Rat IgG2a, κ Isotype Control RUO
Size
0.1 mg
Cat No.
552770
561252 Rev. 3
Antibody Details
Avas 12α1
The Avas 12α1 monoclonal antibody specifically binds to fetal liver kinase 1 (Flk-1) which is also known as CD309. Flk-1 is a receptor protein tyrosine kinase which is closely related to CD117 (c-kit) and CD140a (PDGF Receptor α chain) of the immunoglobulin superfamily. Flk-1, also known as VEGF Receptor-2 (VEGF-R2 or VEGFR2), is a receptor for vascular endothelial growth factor (VEGF). It is expressed, at the mRNA and protein levels, on distinct sets of mesoderm during gastrulation and on endothelial cells in embryonic and adult tissues. In vivo and in vitro studies indicate that Flk-1 is required for the embryonic development of vascular endothelial and hematopoietic cells.
561252 Rev. 3
Format Details
APC-Cy7
APC-Cy7 dye is a part of the BD APC red family of dyes. This tandem fluorochrome is comprised of a Allophycocyanin (APC) donor that has excitation maxima (Ex Max) of 651 nm and an acceptor dye, Cy™7, with an emission maximum (Em Max) at 779 nm. APC-Cy7 can be excited by the Red (627-640 nm) laser and detected using an optical filter centered near 780 nm (e.g., a 760/60 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
APC-Cy7
Red 627-640 nm
651 nm
779 nm
561252 Rev.3
Citations & References
Development References (8)
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Hanahan D. Signaling vascular morphogenesis and maintenance. Science. 1997; 277(5322):48-50. (Biology). View Reference
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Kataoka H, Takakura N, Nishikawa S, et al. Expressions of PDGF receptor alpha, c-Kit and Flk1 genes clustering in mouse chromosome 5 define distinct subsets of nascent mesodermal cells.. Dev Growth Differ. 1997; 39(6):729-40. (Immunogen). View Reference
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Millauer B, Wizigmann-Voos S, Schnurch H, et al. High affinity VEGF binding and developmental expression suggest Flk-1 as a major regulator of vasculogenesis and angiogenesis. Cell. 1993; 72(6):835-846. (Biology). View Reference
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Nishikawa SI, Nishikawa S, Hirashima M, Matsuyoshi N, Kodama H. Progressive lineage analysis by cell sorting and culture identifies FLK1+VE-cadherin+ cells at a diverging point of endothelial and hemopoietic lineages. Development. 1998; 125(9):1747-1757. (Biology). View Reference
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Nishikawa SI, Nishikawa S, Kawamoto H, et al. In vitro generation of lymphohematopoietic cells from endothelial cells purified from murine embryos. Immunity. 1998; 8(6):761-769. (Biology). View Reference
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Quinn TP, Peters KG, De Vries C, Ferrara N, Williams LT. Fetal liver kinase 1 is a receptor for vascular endothelial growth factor and is selectively expressed in vascular endothelium. Proc Natl Acad Sci U S A. 1993; 90(16):7533-7537. (Biology). View Reference
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Shalaby F, Ho J, Stanford WL, et al. A requirement for Flk1 in primitive and definitive hematopoiesis and vasculogenesis. Cell. 1997; 89(6):981-990. (Biology). View Reference
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Shalaby F, Rossant J, Yamaguchi TP, et al. Failure of blood-island formation and vasculogenesis in Flk-1-deficient mice. Nature. 1995; 376(6535):62-66. (Biology). View Reference
561252 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.
