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Alexa Fluor® 647 Mouse Anti-Chondroitin Sulfate
Alexa Fluor® 647 Mouse Anti-Chondroitin Sulfate
Flow cytometric analysis of Chondroitin Sulfate expressed on M21 human melanoma cells. Human M21 cells were stained with either Alexa Fluor® 647 Mouse Anti-Human Chondroitin Sulfate antibody (Cat. No. 562414, solid line histogram) or a Alexa Fluor® 647 mIgG2a, κ Isotype Control (Cat. No. 557715; dashed line histogram). Flow cytometric fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of viable cells. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.
Flow cytometric analysis of Chondroitin Sulfate expressed on M21 human melanoma cells. Human M21 cells were stained with either Alexa Fluor® 647 Mouse Anti-Human Chondroitin Sulfate antibody (Cat. No. 562414, solid line histogram) or a Alexa Fluor® 647 mIgG2a, κ Isotype Control (Cat. No. 557715; dashed line histogram). Flow cytometric fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of viable cells. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.
Product Details
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BD Pharmingen™
CSPG4; NG2; Chondroitin sulfate proteoglycan 4; MCSP; MCSPG; MELCSPG; MSK16
Human (QC Testing)
Mouse BALB/c IgG2a
Extracts of Human M21 Melanoma Cells
Flow cytometry (Routinely Tested)
5 µl
1464
AB_11154050
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 Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was removed.

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. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  4. The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
  5. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  6. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
  7. 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.
  8. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  9. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
562414 Rev. 1
Antibody Details
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9.2.27

The 9.2.27 monoclonal antibody specifically binds to CSPG4 (chondroitin sulfate proteoglycan 4). CSPG4 is also known as MCSP (melanoma chondroitin sulfate proteoglycan) and NG2 (neural/glial antigen 2). The hybridoma secreting the 9.2.27 antibody was generated using a human melanoma cell extract as the immunogen. Tumor cells display a variety of antigens which have been defined by antibodies. Antibodies that specifically react with tumor-associated antigens are useful in understanding the biology of particular tumor types. Proteoglycans including chondroitin sulfate, keratan sulfate, dermatan sulfate and heparan sulfate are major components of the extracellular matrices of many animal cell types. Clone 9.2.27 has been shown to react with human melanoma cells, glioma cells, and proliferating brain endothelial cells. It did not react with fetal melanocytes, neuroblastoma LA-N-1 cells or a variety of carcinoma, lymphoid, and fibroblastoid cell lines. Additionally, neither white nor gray matter from the medulla oblongata, cerebellum or spinal cord of a normal human adult reacted with 9.2.27. Clone 9.2.27 has also been used in functional studies to investigate the role of chondroitin sulfate proteoglycans in human tumor cell systems. It has been shown to block melanoma cell spreading, exhibit antiproliferative effects on glioma cells in vitro and suppress glioma tumor growth in athymic nude mice models.

  

562414 Rev. 1
Format Details
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Alexa Fluor™ 647
Alexa Fluor™ 647 Dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 653-nm and an emission maximum (Em Max) at 669-nm. Alexa Fluor 647 is designed to be excited by the Red laser (627-640 nm) and detected using an optical filter centered near 520-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.
Alexa Fluor™ 647
Red 627-640 nm
653 nm
669 nm
562414 Rev.1
Citations & References
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Development References (7)

  1. Bumol TF, Reisfeld RA. Unique glycoprotein-proteoglycan complex defined by monoclonal antibody on human melanoma cells. Proc Natl Acad Sci U S A. 1982; 79(4):1245-1249. (Biology). View Reference
  2. Bumol TF, Walker LE, Reisfeld RA. Biosynthetic studies of proteoglycans in human melanoma cells with a monoclonal antibody to a core glycoprotein of chondroitin sulfate proteoglycans. J Biol Chem. 1984; 259(20):12733-12741. (Clone-specific: Electron microscopy, Immunofluorescence, Immunoprecipitation). View Reference
  3. Cheresh DA, Honsik CJ, Staffileno LK, Jung G, Reisfeld RA. Disialoganglioside GD3 on human melanoma serves as a relevant target antigen for monoclonal antibody-mediated tumor cytolysis. Proc Natl Acad Sci U S A. 1985; 82(15):5155-5159. (Biology). View Reference
  4. Morgan AC, Galloway DR, Reisfeld RA. Production and characterization of monoclonal antibody to a melanoma specific glycoprotein. Hybridoma. 1981; 1(1):27-36. (Immunogen: Immunoprecipitation). View Reference
  5. Pluschke G, Vanek M, Evans A, et al. Molecular cloning of a human melanoma-associated chondroitin sulfate proteoglycan. Proc Natl Acad Sci U S A. 1996; 93(18):9710-9715. (Clone-specific: Immunoprecipitation). View Reference
  6. Schrappe M, Bumol TF, Apelgren LD. Long-term growth suppression of human glioma xenografts by chemoimmunoconjugates of 4-desacetylvinblastine-3-carboxyhydrazide and monoclonal antibody 9.2.27. Cancer Res. 1992; 52(14):3838-3844. (Clone-specific: Immunofluorescence). View Reference
  7. Spiro RC, Casteel HE, Laufer DM, Reisfeld RA, Harper JR. Post-translational addition of chondroitin sulfate glycosaminoglycans. Role of N-linked oligosaccharide addition, trimming, and processing. J Biol Chem. 1989; 264(3):1779-1786. (Biology). View Reference
View All (7) View Less
562414 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.