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BV421 Mouse Anti-Human Chondroitin Sulfate (NG2)
BV421 Mouse Anti-Human Chondroitin Sulfate (NG2)

Flow cytometric analysis of Chondroitin Sulfate (NG2) expression on Human M21 cells.  Cells from the Human M21 melanoma cell line were preincubated with Human BD Fc Block™ (Cat. No. 564219/564220) and stained with either BD Horizon™ BV421 Mouse IgG1, κ Isotype Control (Cat. No. 562438; dashed line histogram) or BD Horizon™ BV421 Mouse Anti-Human Chondroitin Sulfate (NG2) antibody (Cat. No. 570318/570396; solid line histogram). BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. The fluorescence histogram showing Chondroitin Sulfate (NG2) expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable (7-AAD-negative) cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ Software.

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570318
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BD Horizon™
CSPG4; HMW-MAA; MCSP; MCSPG; NG2
Human (QC Testing)
Mouse IgG1, κ
Human BM stromal cell line established by infection with SV-40
Flow cytometry (Routinely Tested)
5 µl/test
1464
AB_3685661
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.

推奨アッセイ手順

   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.

   For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime BD Horizon Brilliant dyes are used in a multicolor flow cytometry panel.  Fluorescent dye interactions may cause staining artifacts which may affect data interpretation.  The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. When BD Horizon Brilliant Stain Buffer is used in in the multicolor panel, it should also be used in the corresponding compensation controls for all dyes to achieve the most accurate compensation. For the most accurate compensation, compensation controls created with either cells or beads should be exposed to BD Horizon Brilliant Stain Buffer for the same length of time as the corresponding multicolor panel. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).

Product Notices

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. 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).
  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. Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
  7. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  8. For U.S. patents that may apply, see bd.com/patents.

データシート

570318 Rev. 1
抗体の詳細
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7.1

The 7.1 monoclonal antibody specifically recognizes Neural/Glial Antigen 2 (NG2) which is also known as Melanoma chondroitin sulfate proteoglycan (MCSP) and High molecular weight-melanoma associated antigen (HMW-MAA). This ~251 kDa transmembrane proteoglycan is encoded by CSPG4 (Chondroitin sulfate proteoglycan 4). Chondroitin Sulfate (NG2) is expressed by glial cells but not detected on normal marrow hematopoietic and peripheral blood cells. It is highly expressed by cells in some cancers including melanomas, leukemias, sarcomas and carcinomas. Chondroitin Sulfate (NG2) plays important roles in cellular proliferation, migration, and in oncogenic pathways involved in malignant transformation and metastases.  

570318 Rev. 1
フォーマットの詳細
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BV421
The BD Horizon Brilliant Violet™ 421 (BV421) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology based dye has an excitation maximum (Ex Max) of 407-nm and an emission maximum (Em Max) at 423-nm. Driven by BD innovation, BV421 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 420-nm (e.g., a 431/28-nm or 450/50-nm bandpass filter). BV421 is an ideal alternative for V450 as it is approximately ten times brighter with less spillover into the BV510/V500 detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
altImg
BV421
Violet 405 nm
407 nm
423 nm
570318 Rev.1
引用&参考文献
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Development References (7)

  1. Behm FG, Smith FO, Raimondi SC, Pui CH, Bernstein ID. Human homologue of the rat chondroitin sulfate proteoglycan, NG2, detected by monoclonal antibody 7.1, identifies childhood acute lymphoblastic leukemias with t(4;11)(q21;q23) or t(11;19)(q23;p13) and MLL gene rearrangements.. Blood. 1996; 87(3):1134-9. (Immunogen: Flow cytometry). View Reference
  2. Bueno C, Montes R, Martín L, et al. NG2 antigen is expressed in CD34+ HPCs and plasmacytoid dendritic cell precursors: is NG2 expression in leukemia dependent on the target cell where leukemogenesis is triggered?. Leukemia. 2008; 22(8):1475-8. (Clone-specific: Flow cytometry). View Reference
  3. Luo W, Wang X, Kageshita T, Wakasugi S, Karpf AR, Ferrone S. Regulation of high molecular weight-melanoma associated antigen (HMW-MAA) gene expression by promoter DNA methylation in human melanoma cells.. Oncogene. 2006; 25(20):2873-84. (Biology). View Reference
  4. Nakano M, Tamura Y, Yamato M, et al. NG2 glial cells regulate neuroimmunological responses to maintain neuronal function and survival.. Sci Rep. 2017; 7:42041. (Biology). 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. (Biology). View Reference
  6. Tamburini E, Dallatomasina A, Quartararo J, et al. Structural deciphering of the NG2/CSPG4 proteoglycan multifunctionality.. FASEB J. 2019; 33(3):3112-3128. (Biology). View Reference
  7. Wang X, Wang Y, Yu L, et al. CSPG4 in cancer: multiple roles.. Curr Mol Med. 2010; 10(4):419-29. (Biology). View Reference
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570318 Rev. 1

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