BUV395 Mouse Anti-Mouse H-2K[b]/SIINFEKL

BD Horizon™ BUV395 Mouse Anti-Mouse H-2K[b]/SIINFEKL

Name: BUV395 Mouse Anti-Mouse H-2K[b]/SIINFEKL
Brand: BD Horizon™
SKU: 570093

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

(RUO)

Flow cytometric analysis of H-2K[b]/SIINFEKL expression on viable C57BL/6 Mouse splenic leukocytes with or without prior SIINFEKL peptide treatment. Mouse splenic leukocytes were either not pulsed (Left Plot) or were pulsed (Right Plot) 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™ BUV395 Mouse IgG1, κ Isotype Control (Cat. No. 563547; dashed line histograms) or with BD Horizon™ BUV395 Mouse Anti-Mouse H-2K[b]/SIINFEKL antibody (Cat. No. 570012/570093; solid line histograms) at 0.5 µg/test. BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. The fluorescence histogram showing H-2K[b]/SIINFEKL expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable (7-AAD-negative) splenic 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.

Multiparameter flow cytometric analysis of H-2K[b]/SIINFEKL expression on viable C57BL/6 Mouse splenic leukocytes with prior SIINFEKL peptide treatment. Mouse splenic leukocytes were pulsed 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™ BUV395 Mouse IgG1, κ Isotype Control (Cat. No. 563547; Left Plot) or BD Horizon™ BUV395 Mouse Anti-Mouse H-2K[b]/SIINFEKL antibody (Cat. No. 570012/570093; Right Plot) at 0.5 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 plot showing the correlated expression of H-2K[b]/SIINFEKL (or Ig Isotype control staining) versus side-light scatter (SSC-A) signals was derived from gated events with the forward and side-light scatter characteristics of viable (7-AAD-negative) splenic leukocyte populations. 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 unconjugated antibody and free dye were 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.

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).

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.
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

Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) RUO

Size 0.5 mg Cat No. 553142

BUV395 Mouse IgG1, k Isotype Control RUO

Size 50 µg Cat No. 563547

Cell Viability Solution RUO

Size 500 Tests Cat No. 555815

570093 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.

570093 Rev. 1

Format Details

BUV395

The BD Horizon Brilliant™ Ultraviolet 395 (BUV395) Dye is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This base dye is a polymer fluorochrome with an excitation maximum (Ex Max) of 348-nm and an emission maximum (Em Max) at 395-nm. Driven by BD innovation, BUV395 is designed to be excited by the ultraviolet laser (355-nm) and detected using an optical filter centered near 380-nm (e.g., 379/28-nm bandpass filter). BUV395 is the ideal dye when using only one detector on the ultraviolet laser as it spills into no other detectors and no other fluors spill into it. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.

Format: BUV395

Excitation Source: Ultraviolet 355 nm

Excitation Max: 348 nm

Emission Max: 395 nm

570093 Rev.1

Citations & References

View product citations for antibody "570093" 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)

570093 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.