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BV421 Mouse Anti-Rat CD28
BV421 Mouse Anti-Rat CD28

Multiparameter flow cytometric analysis using BD OptiBuild™ BV421 Mouse Anti-Rat CD28 antibody (Cat. No. 742582) on live Lewis rat thymocytes. Flow cytometry was performed using a BD LSRFortessa™ X-20  Flow Cytometer System.

Multiparameter flow cytometric analysis using BD OptiBuild™ BV421 Mouse Anti-Rat CD28 antibody (Cat. No. 742582) on live Lewis rat thymocytes. Flow cytometry was performed using a BD LSRFortessa™ X-20  Flow Cytometer System.

Product Details
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BD OptiBuild™
Cd28; CD28RNA
Rat (Tested in Development)
Mouse BALB/c IgG1, κ
Rat CD28-transfected cell line
Flow cytometry (Qualified)
0.2 mg/ml
AB_2740883
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 BV421 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 421 is covered by one or more of the following US patents: 8,158,444; 8,362,193; 8,575,303; 8,354,239.
  10. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
742582 Rev. 2
Antibody Details
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JJ319

The JJ319 monoclonal antibody specifically recognizes CD28, which is expressed on virtually all T lymphocytes bearing αβ T-cell receptors (TCR), on most γδ TCR-bearing T cells, and on a subset of NK cells. In the thymus, CD28 expression is developmentally regulated during the maturation of αβ TCR-bearing T cells. CD28 is a costimulatory receptor required for activation of T cells; its ligands include CD80 (B7-1) and CD86 (B7-2). Soluble JJ319 mAb costimulates the proliferative responses and IL-2 production of CD4+ and CD8+ T cells activated by anti-αβ-TCR mAb R73 (Cat. no. 554910). The alternate anti-rat CD28 mAb JJ316 (Cat. No. 554992) is capable of directly stimulating T cells in vitro and in vivo.

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.

742582 Rev. 2
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
742582 Rev.2
Citations & References
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Development References (4)

  1. Bluestone JA. New perspectives of CD28-B7-mediated T cell costimulation. Immunity. 1995; 2(6):555-559. (Biology). View Reference
  2. Mitnacht R, Tacke M, Hunig T. Expression of cell interaction molecules by immature rat thymocytes during passage through the CD4+8+ compartment: developmental regulation and induction by T cell receptor engagement of CD2, CD5, CD28, CD11a, CD44 and CD53. Eur J Immunol. 1995; 25(2):328-332. (Biology). View Reference
  3. Tacke M, Clark GJ, Dallman MJ, Hunig T. Cellular distribution and costimulatory function of rat CD28. Regulated expression during thymocyte maturation and induction of cyclosporin A sensitivity of costimulated T cell responses by phorbol ester. J Immunol. 1995; 154(10):5121-5127. (Immunogen). View Reference
  4. Tacke M, Hanke G, Hanke T, Hunig T. CD28-mediated induction of proliferation in resting T cells in vitro and in vivo without engagement of the T cell receptor: evidence for functionally distinct forms of CD28. Eur J Immunol. 1997; 27(1):239-247. (Clone-specific). View Reference
View All (4) View Less
742582 Rev. 2

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.