RY586 Mouse Anti-Mouse H-2Kb/SIINFEKL

BD Horizon™ RY586 Mouse Anti-Mouse H-2Kb/SIINFEKL

Name: RY586 Mouse Anti-Mouse H-2Kb/SIINFEKL
Brand: RY586 Mouse Anti-Mouse H-2Kb/SIINFEKL
SKU: 570099

Clone 25-D1.16.rMAb (also known as 25-D1.16) (RUO)

Flow cytometric analysis of H-2Kb/SIINFEKL expression on viable C57BL/6 Mouse splenic leukocytes with or without prior SIINFEKL peptide treatment. Mouse splenic leukocytes were either not pulsed (Left Panel) or pulsed (Right Panel) with SIINFEKL peptide for 2 hours and were then preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) [Cat. No. 553142]. The cells were stained with either BD Horizon™ RY586 Mouse IgG1, κ Isotype Control (Cat. No. 568097; dashed line histograms) or BD Horizon™ RY586 Mouse Anti-Mouse H-2Kb/SIINFEKL antibody (Cat. No. 570018/570099; solid line histograms) at 0.5 µg/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. The fluorescence histograms showing H-2Kb/SIINFEKL expression (or Ig Isotype control expression) were derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) leukocytes. 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: H-2Kb/SIINFEKL; H-2Kb:SIINFEKL; SIINFEKL-bound H-2Kb

Reactivity: Mouse (QC Testing)

Isotype: Mouse IgG1, κ

Immunogen: SIINFEKL-pulsed RMA-S cells

Application: Flow cytometry (Routinely Tested)

Concentration: 0.2 mg/ml

Entrez Gene ID: 14972

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

Regulatory Status: RUO

Preparation And Storage

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 unreacted dye was removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.

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.

Product Notices

Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Since applications vary, each investigator should titrate the reagent to obtain optimal results.
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.
CF™ is a trademark of Biotium, Inc.
Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
For U.S. patents that may apply, see bd.com/patents.

Companion Products

Stain Buffer (FBS) RUO

Size 500 mL Cat No. 554656

Stain Buffer (BSA) RUO

Size 500 mL Cat No. 554657

Lysing Buffer RUO

Size 100 mL Cat No. 555899

RY586 Mouse IgG1, κ Isotype Control RUO

Size 50 µg Cat No. 568097

DAPI Solution RUO

Size 1 mg Cat No. 564907

570099 Rev. 1

Antibody Details

25-D1.16.rMAb

25-D1.16.rMAb is a recombinant monoclonal antibody derived from 25-D1.16 hybridoma cells. The 25-D1.16.rMAb specifically recognizes the ovalbumin (OVA)-derived peptide SIINFEKL (amino acid residues 257-264 of OVA) bound to the MHC class l antigen, H-2Kb. It does not bind to either unbound H-2Kb or H-2Kb bound to an irrelevant peptide. The 25-D1.16 antibody has been found useful in a variety of in vitro and in vivo experimental model systems to study the nature of antigen-presenting cells that can present the SIINFEKL peptide in an MHC class I-restricted fashion to T cells. The 25-D1.16 monoclonal antibody can reportedly inhibit the T cell response to H-2Kb-SIINFEKL and be used for immunofluorescent or immunohistochemical staining of H-2Kb-SIINFEKL-positive cells.

570099 Rev. 1

Format Details

RY586

The BD Horizon RealYellow™ 586 (RY586) Dye is part of the BD family of yellow-green dyes. It is a small organic fluorochrome with an excitation maximum (Ex Max) at 565-nm and an emission maximum (Em Max) at 586-nm. Driven by BD innovation, RY586 can be used on both spectral and conventional cytometers and is designed to be excited by the Yellow-Green laser (561-nm) with minimal excitation by the 488-nm Blue laser. For conventional instruments equipped with a Yellow-Green laser (561-nm), RY586 can be used as an alternative to PE and we recommend using an optical filter centered near 586-nm (eg, a 586/15-nm bandpass filter). For spectral instruments equipped with a Yellow-Green laser (561-nm), it can be used in conjunction with PE. Compared to PE, RY586 is similar in brightness, minimal spillover into Blue detectors, and increased spillover into the 610/20-nm (PE-CF594) detector. Please ensure that your instrument configuration (lasers and optical filters) is appropriate for this dye.

Format: RY586

Excitation Source: Yellow-Green 561 nm

Excitation Max: 564 nm

Emission Max: 586 nm

570099 Rev.1

Citations & References

View product citations for antibody "570099" on CiteAb

Development References (4)

Dolan BP, Li L, Takeda K, Bennink JR, Yewdell JW. Defective ribosomal products are the major source of antigenic peptides endogenously generated from influenza A virus neuraminidase. J Immunol. 2010; 184(3):1419-1424. (Clone-specific: Flow cytometry, Fluorescence microscopy, Immunofluorescence). View Reference
Hervé J, Dubreil L, Tardif V, et al. β2-Adrenoreceptor agonist inhibits antigen cross-presentation by dendritic cells.. J Immunol. 2013; 190(7):3163-71. (Clone-specific: Flow cytometry, Fluorescence microscopy, Immunofluorescence). View Reference
Mareeva T, Wanjalla C, Schnell MJ, Sykulev Y. A novel composite immunotoxin that suppresses rabies virus production by the infected cells.. J Immunol Methods. 2010; 353(1-2):78-86. (Clone-specific: Cytotoxicity, Flow cytometry). View Reference
Porgador A, Yewdell JW, Deng Y, Bennink JR, Germain RN. Localization, quantitation, and in situ detection of specific peptide-MHC class I complexes using a monoclonal antibody.. Immunity. 1997; 6(6):715-26. (Immunogen: Flow cytometry, Fluorescence microscopy, Immunofluorescence, Immunohistochemistry). View Reference

View All (4)

570099 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.