BV480 Rat Anti-Mouse CD8a

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BD OptiBuild™ BV480 Rat Anti-Mouse CD8a

Name: BV480 Rat Anti-Mouse CD8a
Brand: BD OptiBuild™
SKU: 752641

Clone 5H10-1

(RUO)

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Product Details

Brand: BD OptiBuild™

Alternative Name: Cd8a; CD8 antigen, alpha chain; CD8a; CD8α; Ly-2; Lyt2; Lyt-2

Reactivity: Mouse (Tested in Development)

Isotype: Rat SD, also known as Sprague-Dawley (outbred) IgG2b, λ

Immunogen: Concanavalin A-stimulated BALB/c Splenic T Cell Blasts

Application: Flow cytometry (Qualified)

Concentration: 0.2 mg/ml

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 that minimize unconjugated dye and antibody.

Recommended Assay Procedures

BD™ CompBeads can be used as surrogates to assess fluorescence spillover (Compensation). When fluorochrome conjugated antibodies are bound to 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 CompBead to ensure that BD Comp beads are appropriate for your specific cellular application.

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. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).

For Immunofluorescence Applications:

The use of a mounting reagent (eg, ProLong® Gold) is highly recommended to maximize the photostability of BV480. For confocal microscopy systems, a 440 nm laser is the optimal excitation source and the recommended emission filter is a 485/20 nm bandpass filter.

For epifluorescence microscopes with broad spectrum excitation sources, the recommended excitation and emission filters are 445/20 nm and 485/20 nm bandpass filters, respectively. For specific multicolor imaging applications, the exact filter configurations should be optimized by the end user. For additional instrument/filter configuration information, please visit http://www.bdbiosciences.com/research/cellularimaging.

Product Notices

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.
Researchers should determine the optimal concentration of this reagent for their individual applications.
An isotype control should be used at the same concentration as the antibody of interest.
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.
For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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).
BD Horizon Brilliant Violet 480 is covered by one or more of the following US patents: 8,575,303; 8,354,239.
ProLong® is a registered trademark of Thermo Fisher Scientific, Inc. Waltham, MA.
BD Horizon Brilliant Violet 480 is covered by one or more of the following US patents: 8,575,303; 8,354,239.

Companion Products

Brilliant Stain Buffer RUO

Size 1000 Tests Cat No. 566349

Stain Buffer (FBS) RUO

Size 500 mL Cat No. 554656

Brilliant Stain Buffer RUO

Size 100 Tests Cat No. 563794

Stain Buffer (BSA) RUO

Size 500 mL Cat No. 554657

Brilliant Stain Buffer Plus RUO

Size 1000 Tests Cat No. 566385

Lysing Buffer RUO

Size 100 mL Cat No. 555899

View Details

752641 Rev. 1

Antibody Details

5H10-1

The 5H10-1 monoclonal antibody specifically recognizes CD8a which is also known as CD8 alpha (CD8α), Ly-2 or Lyt-2. CD8a is an ~34 kDa single pass type I transmembrane glycoprotein that is encoded by Cd8a (CD8 antigen, alpha chain) which belongs to the immunoglobulin superfamily (IgSF). CD8a is comprised of an extracellular region that contains an IgV-like domain followed by a transmembrane region and cytoplasmic tail with a Lck tyrosine kinase binding motif. CD8a is expressed on the cell surface as either a disulfide bond-linked homodimer (CD8αα) or a heterodimer (CD8αβ) when disulfide bonded to CD8b (also known as, CD8 beta/CDβ, Ly-3, or Lyt-3), another type I transmembrane glycoprotein. CD8aβ is expressed on most thymocytes and a subpopulation of mature peripheral TCR αβ T cells including some intraepithelial lymphocytes (IELs). CD8αα is expressed on IELs which include either TCR αβ or TCR γδ T cells as well as on other leucocytes, such as, subsets of NK cells and dendritic cells (DCs). CD8αβ serves as a co-receptor for antigen-stimulated CD8+ T cells by binding to the same peptide:MHC class I complex as the T cell receptor and by helping to trigger the subsequent signaling cascade through Lck activation upon antigen recognition. Through high-affinity binding to nonclassical MHC class-Ib molecules, CD8αα can reportedly function less efficiently as a co-receptor and even repress antigen-stimulated CD8+ T cell responses.

The antibody was conjugated to BD Horizon BV480 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 436-nm and Em Max at 478-nm, BD Horizon BV480 can be excited by the violet laser and detected in the BD Horizon BV510 (525/40-nm) filter set. BV480 has less spillover into the BV605 detector and, in general, is brighter than BV510.

752641 Rev. 1

Format Details

BV480

The BD Horizon Brilliant Violet™ 480 (BV480) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology fluorochrome has an excitation maximum (Ex Max) of 440-nm and an emission maximum (Em Max) of 479-nm. Driven by BD innovation, BV480 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 480-nm (e.g., a 525/50 bandpass filter). The increased fluorescence intensity of BV480 and narrower emission spectra, make it a good alternative for BV510 or V500. Due to its excitation profile, BV480 will also has less cross-laser excitation with the UV laser, resulting in less spillover into UV channels compared to BV510. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.

Format: BV480

Excitation Source: Violet 405 nm

Excitation Max: 440 nm

Emission Max: 479 nm

752641 Rev.1

Citations & References

Development References (7)

Barclay NA, Brown MH, Birkeland ML, et al, ed. The Leukocyte Antigen FactsBook. San Diego, CA: Academic Press; 1997.
LeFrancois L. Extrathymic differentiation of intraepithelial lymphocytes: generation of a separate and unequal T-cell repertoire. Immunol Today. 1991; 12(12):436-438. (Biology). View Reference
Probst HC, McCoy K, Okazaki T, Honjo T, van den Broek M. Resting dendritic cells induce peripheral CD8+ T cell tolerance through PD-1 and CTLA-4.. Nat Immunol. 2005; 6(3):280-6. (Clone-specific: Flow cytometry). View Reference
Ren H, Ferguson BJ, Maluquer de Motes C, Sumner RP, Harman LE, Smith GL. Enhancement of CD8(+) T-cell memory by removal of a vaccinia virus nuclear factor-κB inhibitor.. Immunology. 2015; 145(1):34-49. (Clone-specific: Flow cytometry). View Reference
Sydora BC, Brossay L, Hagenbaugh A, Kronenberg M, Cheroutre H. TAP-independent selection of CD8+ intestinal intraepithelial lymphocytes. J Immunol. 1996; 156(11):4209-4216. (Biology). View Reference
Takahashi K, Nakata M, Tanaka T, et al. CD4 and CD8 regulate interleukin 2 responses of T cells. Proc Natl Acad Sci U S A. 1992; 89(12):5557-5561. (Immunogen: Functional assay, Immunofluorescence, Inhibition). View Reference
Vremec D, Zorbas M, Scollay R, et al. The surface phenotype of dendritic cells purified from mouse thymus and spleen: investigation of the CD8 expression by a subpopulation of dendritic cells. J Exp Med. 1992; 176(1):47-58. (Biology). View Reference

View All (7)

752641 Rev. 1

Please refer to Support Documents for Quality Certificates

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.