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BV786 Rat Anti-Mouse CD274
BV786 Rat Anti-Mouse CD274

Multiparameter flow cytometric analysis using BD OptiBuild™ BV786 Rat Anti-Mouse CD274 antibody (Cat. No. 741014; right plot)) on live mouse  splenocytes stimulated by plate-bound NA/LE Hamster Anti-Mouse CD3e antibody for 3 days, with FMO control (left plot).  Flow cytometry was performed using a BD LSRFortessa™ X-20  Flow Cytometer System.

Multiparameter flow cytometric analysis using BD OptiBuild™ BV786 Rat Anti-Mouse CD274 antibody (Cat. No. 741014; right plot)) on live mouse  splenocytes stimulated by plate-bound NA/LE Hamster Anti-Mouse CD3e antibody for 3 days, with FMO control (left plot).  Flow cytometry was performed using a BD LSRFortessa™ X-20  Flow Cytometer System.

Product Details
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BD OptiBuild™
B7-H1, PD-L1; PD1L1; Programmed death ligand 1
Mouse (Tested in Development)
Rat SD, also known as Sprague-Dawley (outbred) IgG2a, λ
DBA/2 mouse lymphoma L5178Y transfected with Pdcd1lg1 cDNA
Flow cytometry (Qualified)
0.2 mg/ml
60533
AB_2740636
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 BV786 under optimal conditions that minimize unconjugated dye and antibody.

Recommended Assay Procedures

For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes (including BD OptiBuild Brilliant reagents) 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. This antibody was developed for use in flow cytometry.
  2. The production process underwent stringent testing and validation to assure that it generates a high-quality conjugate with consistent performance and specific binding activity. However, verification testing has not been performed on all conjugate lots.
  3. Researchers should determine the optimal concentration of this reagent for their individual applications.
  4. An isotype control should be used at the same concentration as the antibody of interest.
  5. 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.
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  8. 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.
  9. BD Horizon Brilliant Violet 786 is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,227,187; 8,455,613; 8,575,303; 8,354,239.
  10. Cy is a trademark of Amersham Biosciences Limited.
741014 Rev. 2
Antibody Details
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MIH5

The MIH5 monoclonal antibody specifically binds to CD274, also known as B7-H1 or PDL1, a 43-kDa glycoprotein encoded by the Pdcd1lg1 gene of the B7 family of the Ig superfamily. Pdcd1lg1 mRNA is expressed in more tissues than other members of the B7 family; transcripts are found in lymphoid tissues and many, but not all, non-lymphoid tissues. The protein has been detected at low levels on resting peripheral T and B lymphocytes, macrophages, and dendritic cells. B7-H1 mRNA and protein expression are upregulated upon activation of T and B cells, macrophages, dendritic cells, and epidermal keratinocytes by a variety of stimulatory factors. B7-H1's receptor, PD-1, contains an ITIM (Immunoreceptor Tyrosine-based Inhibitory Motif) on its intracytoplasmic region and is expressed on activated B and T lymphocytes, suggesting that B7-H1-PD-1 interaction may be involved in the negative regulation of immune responses. The second PD-1 ligand, B7-DC (PD-L2), is also a member of the B7 family of the Ig superfamily. Furthermore, B7-H1 may participate in positive immunoregulation, or costimulation of T cells, through an additional receptor, which is not PD-1 and distinct from the alternate receptor for B7-DC. The MIH5 antibody blocks the binding of PD-1-Ig to B7-H1 transfectants.

The antibody was conjugated to BD Horizon™ BV786 which is part of the BD Horizon Brilliant™ Violet family of dyes. This dye is a tandem fluorochrome of BD Horizon BV421 with an Ex Max of 405-nm and an acceptor dye with an Em Max at 786-nm.  BD Horizon BV786 can be excited by the violet laser and detected in a filter used to detect Cy™7-like dyes (eg, 780/60-nm filter).

741014 Rev. 2
Format Details
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BV786
The BD Horizon Brilliant Violet™ 786 (BV786) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This tandem fluorochrome is comprised of a BV421 donor with an Ex Max of 407-nm and an acceptor dye with an Em Max at 786-nm.  BV786, driven by BD innovation, is designed to be excited by the violet laser and detected using a filter, centered near 785 nm (e.g. 780/60 nm bandpass filter).  Please ensure that your instrument’s configurations (lasers and filters) are appropriate for this dye.
altImg
BV786
Violet 405 nm
407 nm
786 nm
741014 Rev.2
Citations & References
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Development References (9)

  1. Ansari MJ, Salama AD, Chitnis T, et al. The programmed death-1 (PD-1) pathway regulates autoimmune diabetes in nonobese diabetic (NOD) mice. J Exp Med. 2003 July; 198(1):63-69. (Biology). View Reference
  2. Carreno BM, Collins M. The B7 family of ligands and its receptors: New pathways for costimulation and inhibition of immune responses. Annu Rev Immunol. 2002; 20:29-53. (Biology). View Reference
  3. Dong H, Chen L. B7-H1 pathway and its role in the evasion of tumor immunity. J Mol Med. 2003 May; 81(5):281-287. (Biology). View Reference
  4. Hessel EM, Chu M, Lizcano JO, et al. Immunostimulatory oligonucleotides block allergic airway inflammation by inhibiting Th2 cell activation and IgE-mediated cytokine induction. J Exp Med. 2005; 202(11):1563-1573. (Clone-specific: Flow cytometry). View Reference
  5. Liu X, Gao JX, Wen J, et al. B7DC/PDL2 promotes tumor immunity by a PD-1-independent mechanism. J Exp Med. 2003; 197:1721-1730. (Biology). View Reference
  6. Tamura H, Dong H, Zhu G, et al. B7-H1 costimulation preferentially enhances CD28-independent T-helper cell function. Blood. 2001; 97(6):1809-1816. (Biology). View Reference
  7. Tsushima F, Iwai H, Otsuki N, et al. Preferential contribution of B7-H1 to programmed death-1-mediated regulation of hapten-specific allergic inflammatory responses. Eur J Immunol. 2003; 33(10):2773-2782. (Immunogen: Blocking, Enhancement, Functional assay, Immunofluorescence, Immunohistochemistry, Inhibition, In vivo exacerbation). View Reference
  8. Wang S, Bajorath J, Flies DB, Dong H, Honjo T, Chen L. Molecular modeling and functional mapping of B7-H1 and B7-DC uncouple costimulatory function from PD-1 interaction. J Exp Med. 2003; 197:1083-1091. (Biology). View Reference
  9. Yamazaki T, Akiba H, Iwai H, et al. Expression of programmed death 1 ligands by murine T cells and APC. J Immunol. 2002; 169(10):5538-5545. (Biology). View Reference
View All (9) View Less
741014 Rev. 2

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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.

Refer to manufacturer's instructions for use and related User Manuals and Technical Data Sheets before using this product 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.