BUV737 Rat Anti-Mouse CD274

This product is the replacement for 741877.

BD Horizon™ BUV737 Rat Anti-Mouse CD274

Name: BUV737 Rat Anti-Mouse CD274
Brand: BD Horizon™
SKU: 568361

Clone MIH5

(RUO)

Multicolor flow cytometric analysis of CD274 expression on nonactivated and activated Mouse splenocytes. Mouse splenic leucocytes were either not activated (Upper Plots) or activated (Bottom Plots) by culture with plate-bound Purified NA/LE Hamster Anti-Mouse CD3 antibody (Cat. No. 553057) for 3 days (37°C). The cells were harvested, washed and preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) [Cat. No. 553141/553142]. The cells were then stained with PE Rat Anti-Mouse CD4 antibody (Cat. No. 553730) and with either BD Horizon™ BUV737 Rat IgG2a, κ Isotype Control (Cat. No. 612760; Left Plots) or BD Horizon™ BUV737 Rat Anti-Mouse CD274 antibody (Cat. No. 568361; Right Plots) at 0.25 µg/test. BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. The bivariate pseudocolor density plots showing the correlated expression of CD274 (or Ig Isotype control staining) versus CD4 were derived from gated events with the forward and side light- scatter characteristics of viable (7-AAD-negative) leucocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.

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Product Details

Brand: BD Horizon™

Alternative Name: B7-H1, PD-L1; PD1L1; Programmed death ligand 1

Reactivity: Mouse (QC Testing)

Isotype: Rat SD, also known as Sprague-Dawley (outbred) IgG2a, λ

Immunogen: DBA/2 mouse lymphoma L5178Y transfected with Pdcd1lg1 cDNA

Application: Flow cytometry (Routinely Tested)

Concentration: 0.2 mg/ml

Entrez Gene ID: 60533

Storage Buffer: Aqueous buffered solution containing ≤0.09% sodium azide.

Regulatory Status: 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 the dye under optimum conditions and unconjugated antibody and free dye were removed.

Recommended Assay Procedures

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).

Note: When using high concentrations of antibody, background binding of this dye to erythroid cell subsets (mature erythrocytes and precursors) has been observed. For researchers studying these cell populations, or in cases where light scatter gating does not adequately exclude these cells from the analysis, this background may be an important factor to consider when selecting reagents for panel(s).

Product Notices

Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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.
Since applications vary, each investigator should titrate the reagent to obtain optimal results.
For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
An isotype control should be used at the same concentration as the antibody of interest.
BD Horizon Brilliant Ultraviolet 737 is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,227,187; 8,575,303; 8,354,239.
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.
Please refer to http://regdocs.bd.com to access safety data sheets (SDS).

Companion Products

Stain Buffer (FBS) RUO

Size 500 mL Cat No. 554656

Stain Buffer (BSA) RUO

Size 500 mL Cat No. 554657

Brilliant Stain Buffer RUO

Size 1000 Tests Cat No. 566349

Brilliant Stain Buffer Plus RUO

Size 1000 Tests Cat No. 566385

BUV737 Rat IgG2a, κ Isotype Control RUO

Size 50 µg Cat No. 612760

Cell Viability Solution RUO

Size 500 Tests Cat No. 555815

View Details

568361 Rev. 1

Antibody Details

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.

568361 Rev. 1

Format Details

BUV737

The BD Horizon Brilliant™ Ultraviolet 737 (BUV737) Dye is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This tandem fluorochrome is comprised of a BUV395 donor with an excitation maximum (Ex Max) of 350-nm and an acceptor dye with an emission maximum (Em Max) at 735-nm. BUV737, driven by BD innovation, is designed to be excited by the ultraviolet laser (355-nm) and detected using an optical filter centered near 740-nm (e.g., 740/35 bandpass filter). The acceptor dye can be excited by the Red (628–640nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.

Format: BUV737

Excitation Source: Ultraviolet 355 nm

Excitation Max: 350 nm

Emission Max: 735 nm

568361 Rev.1

Citations & References

View product citations for antibody "568361" on CiteAb

Development References (9)

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
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
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
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
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
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
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
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
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)

568361 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.