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BV421 Mouse Anti-Human CD276
BV421 Mouse Anti-Human CD276
CD276 expression on human immature dendritic cells. Human PBMC were isolated using Ficoll-Paque density gradient medium and cultured for three hours. The non-adherent fraction was removed, and the remaining adherent cells were cultured with Recombinant Human IL-4 (Cat. No. 554605) and Recombinant Human GM-CSF (Cat. No. 550068) for five days. The cells were harvested and stained with either BD Horizon™ BV421 Mouse Anti-Human CD276 (Cat. No. 565829, solid-line histogram) or BD Horizon™ BV421 Mouse IgG1, κ Isotype Control (Cat. No. 562438, dashed-line histogram). Samples were washed  with BD Pharmingen™ Stain Buffer (FBS) and data was acquired. The histograms were derived from gated events with the forward and side light-scatter characteristics of viable dendritic cells. Flow cytometry was performed on a BD LSRFortessa™ Flow Cytometer System. Overlay plots were generated using FlowJo™ Software.
BV421 Mouse Anti-Human CD276
Immunofluorescent staining of CD276 on human placenta (Left Panel) and tonsil (Right Panel). The acetone-fixed frozen sections were blocked using an Avidin/Biotin Blocking Kit (Vector Laboratories, Cat. No. SP-2001) as recommended by the manufacturer. The sections were then stained with BD Horizon™ BV421 Mouse Anti-Human CD276 (Cat. No. 565829, pseudocolored green, both panels) and Alexa Fluor®647 Mouse Anti-Human CD3 (Cat. No. 557706, pseudocolored red, right panel only). LEFT: CD276 expression on trophoblast and Hofbauer cells can be observed in the placenta. RIGHT: CD276 expression (green) can be observed on some B lymphocytes, dendritic cells and macrophages surrounded by paracortical CD3-positive cells (red). The photographs were performed on a standard  epifluorescence microscope. Original magnification, 20×.
CD276 expression on human immature dendritic cells. Human PBMC were isolated using Ficoll-Paque density gradient medium and cultured for three hours. The non-adherent fraction was removed, and the remaining adherent cells were cultured with Recombinant Human IL-4 (Cat. No. 554605) and Recombinant Human GM-CSF (Cat. No. 550068) for five days. The cells were harvested and stained with either BD Horizon™ BV421 Mouse Anti-Human CD276 (Cat. No. 565829, solid-line histogram) or BD Horizon™ BV421 Mouse IgG1, κ Isotype Control (Cat. No. 562438, dashed-line histogram). Samples were washed  with BD Pharmingen™ Stain Buffer (FBS) and data was acquired. The histograms were derived from gated events with the forward and side light-scatter characteristics of viable dendritic cells. Flow cytometry was performed on a BD LSRFortessa™ Flow Cytometer System. Overlay plots were generated using FlowJo™ Software.
Immunofluorescent staining of CD276 on human placenta (Left Panel) and tonsil (Right Panel). The acetone-fixed frozen sections were blocked using an Avidin/Biotin Blocking Kit (Vector Laboratories, Cat. No. SP-2001) as recommended by the manufacturer. The sections were then stained with BD Horizon™ BV421 Mouse Anti-Human CD276 (Cat. No. 565829, pseudocolored green, both panels) and Alexa Fluor®647 Mouse Anti-Human CD3 (Cat. No. 557706, pseudocolored red, right panel only). LEFT: CD276 expression on trophoblast and Hofbauer cells can be observed in the placenta. RIGHT: CD276 expression (green) can be observed on some B lymphocytes, dendritic cells and macrophages surrounded by paracortical CD3-positive cells (red). The photographs were performed on a standard  epifluorescence microscope. Original magnification, 20×.
Product Details
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BD Horizon™
B7-H3, B7H3, 4Ig-B7-H3, B7RP-2
Human (QC Testing)
Mouse BALB/c IgG1, κ
Human Monocyte-Derived Dendritic Cells
Flow cytometry (Routinely Tested), Immunofluorescence (Tested During Development)
0.2 mg/ml
VIII 80650
80381
AB_2739369
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 with BD Horizon BV421 under optimum conditions, and unconjugated antibody and free BD Horizon BV421 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 of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794).

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  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. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
  6. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  7. Ficoll-Paque is a trademark of Amersham Biosciences Limited.
  8. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  9. FlowJo is a trademark of Tree Star Inc.
  10. An isotype control should be used at the same concentration as the antibody of interest.
565829 Rev. 1
Antibody Details
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7-517

The 7-517 monoclonal antibody specifically binds to CD276, also known as B7-H3 (B7 homolog 3). CD276 is a type I transmembrane glycoprotein and member of the B7 family of immunoregulatory proteins. Although the CD276 gene transcript is expressed in nearly all normal tissues, the protein is preferentially expressed in tumors, tumor cell lines, sinonasal epithelial cells, and extravillous trophoblast and Hofbauer cells of the placenta. CD276 protein expression can be induced by activation of T and B lymphocytes, natural killer (NK) cells and antigen-presenting cells. The roles of CD276 in the regulation of immune responses, especially to cancer, are under investigation.  

The antibody was conjugated to BD Horizon BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 407-nm and Em Max at 421-nm, BD Horizon BV421 can be excited by the violet laser and detected in the standard Pacific Blue™ filter set (eg, 450/50-nm filter). BD Horizon BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue conjugates.

565829 Rev. 1
Format Details
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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.
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BV421
Violet 405 nm
407 nm
423 nm
565829 Rev.1
Citations & References
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Development References (6)

  1. Sallusto F, Lanzavecchia A. Efficient presentation of soluble antigen by cultured human dendritic cells is maintained by granulocyte/macrophage colony-stimulating factor plus interleukin 4 and downregulated by tumor necrosis factor alpha.. J Exp Med. 1994; 179(4):1109-18. (Methodology). View Reference
  2. Sharpe AH, Freeman GJ. The B7-CD28 superfamily.. Nat Rev Immunol. 2002; 2(2):116-26. (Biology). View Reference
  3. Steinberger P. B7-H3 ameliorates GVHD.. Blood. 2015; 125(21):3219-21. (Biology). View Reference
  4. Steinberger P1, Majdic O, Derdak SV, et al. Molecular characterization of human 4Ig-B7-H3, a member of the B7 family with four Ig-like domains.. J Immunol. 2004; 172(4):2352-2359. (Immunogen: Flow cytometry, Western blot). View Reference
  5. Sun J, Liu C, Gao L, et al. Correlation between B7-H3 expression and rheumatoid arthritis: A new polymorphism haplotype is associated with increased disease risk.. Clin Immunol. 2015; 159(1):23-32. (Biology). View Reference
  6. Sun M, Richards S, Prasad DV, Mai XM, Rudensky A, Dong C. Characterization of mouse and human B7-H3 genes.. J Immunol. 2002; 168(12):6294-7. (Biology). View Reference
View All (6) View Less
565829 Rev. 1

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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.