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Each lot of this antibody is quality control tested by immunofluorescent staining with flow cytometric analysis. For immunofluorescent staining, the suggested use of this reagent is ≤1.0 µg per million cells in 100 µl volume. It is recommended that the reagent be titrated for optimal performance for each application.
COA:
Application Notes:
The 25-D1.16 monoclonal antibody specifically reacts with ovalbumin-derived peptide SIINFEKL bound to H-2Kb of MHC class I, but not with unbound H-2Kb or H-2Kb bound with an irrelevant peptide. Additional reported applications (for relevant formats) include: Western Blotting1,3, immunofluorescence microscopy2,3, immunohistochemical staining of frozen tissue sections3 , and inhibition of T cell response to H-2Kb-SIINFEKL in vitro.
C57BL/6 mouse splenocytes were pulsed with SIINFELK for 2 hours and then stained with purified anti-mouse SIINFEKL bound H-2Kb (clone 25-D1.16) (filled histogram) or mouse IgG1, κ isotype control (open histogram), followed by anti-mouse IgG PE
This antibody has been proven to be very useful in tracking the quantity and localization of these specific antigen-presenting cells (APC) in vivo.
Other Names:
MHC class I molecule Kb bound to the peptide SIINFEKL (Kb-SIINFEKL) H-2Kb, OVA agonist peptide (OVAp), OVA-Kb, OVA257-264 (SIINFEKL)-H-2kb, OVA257-264-H-2kb
Structure:
OVA peptide257-264 and H-2Kb complex
Distribution:
SIINFEKL bond H-2Kb bearing cells
Antigen References:
1. Mareeva T, et al. 2010. J. Immunol. Methods 353:78. 2. Mareeva T, et al. 2008. J. Biol. Chem. 283:29053. 3. Deng Y, et al. 1998. J. Immunol. 161:1677.
Purified anti-mouse H-2Kb bound to SIINFEKL, 25-D1.16, Purified, MHC class I molecule Kb bound to the peptide SIINFEKL (Kb-SIINFEKL) H-2Kb, OVA agonist peptide (OVAp), OVA-Kb, OVA257-264 (SIINFEKL)-H-2kb, OVA257-264-H-2kb
*These products may be covered by one or more Limited Use Label Licenses (see the BioLegend Catalog or our website, www.biolegend.com/terms). BioLegend products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products, reverse engineer functionally similar materials, or to provide a service to third parties without written approval of BioLegend. By use of these products you accept the terms and conditions of all applicable Limited Use Label Licenses. Unless otherwise indicated, these products are for research use only and are not intended for human or animal diagnostic, therapeutic or commercial use.
This data display is provided for general comparisons between formats. Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors. If you need assistance with selecting the best format contact our expert technical support team.
PE anti-mouse H-2Kb bound to SIINFEKL
C57BL/6 mouse splenocytes were pulsed with (filled histogram) or without (cyan open histogram) SIINFELK for 2 hours and then stained with 25-D1.16 PE or mouse IgG1, κ PE isotype control (broken line open histogram).
Purified anti-mouse H-2Kb bound to SIINFEKL
C57BL/6 mouse splenocytes were pulsed with SIINFELK for 2 hours and then stained with purified anti-mouse SIINFEKL bound H-2Kb (clone 25-D1.16) (filled histogram) or mouse IgG1, κ isotype control (open histogram), followed by anti-mouse IgG PE
APC anti-mouse H-2Kb bound to SIINFEKL
C57BL/6 mouse splenocytes were pulsed with or without SIINFEKL for 2 hrs, and then stained with anti-mouse SIINFEKL bound H-2Kb (clone 25-D1.16) APC (purple histogram indicates pulsed cells and cyan open histogram indicates non-pulsed cells) or mouse IgG1, κ APC isotype control (black open histogram).
CHO cells expressing inducible single chain H-2Kb-SIINFEKL construct were induced by the addition of doxycycline overnight. The negative control is non-doxycycline treated cells, same staining procedure. Stained live cells in PBS+1%NGS+ APC-H-2Kb-SIINFEKL, clone 25-D1.16 (1:200 dilution, 10 µg/ml) for 1h RT, then fixed with PFA 4% RT 10 min and counterstained with DAPI 1 mg/ml RT 5 min. Image courtesy of Dr. Javier Casas, The Scripps Research Institute.
PE/Cy7 anti-mouse H-2Kb bound to SIINFEKL
C57BL/6 mouse splenocytes were pulsed with or without SIINFELK for 2 hours, and then stained with anti-mouse SIINFEKL bound H-2Kb (clone 25-D1.16) PE/Cy7 (purple filled histogram indicates the pulsed cells and cyan open histogram indicates non-pulsed cells) or mouse IgG1, κ PE/Cy7 isotype control (black open histogram).
PerCP/Cy5.5 anti-mouse H-2Kb bound to SIINFEKL
C57BL/6 mouse splenocytes were pulsed with or without SIINFELKfor 2 hours, and then stained with anti-mouse SIINFEKL bound H-2Kb (clone 25-D1.16) PerCP/Cy5.5 (purple filled histogram indicates the pulsed cells and cyan open histogram indicates non-pulsed cells) or mouse IgG1, κ PerCP/Cy5.5 isotype control (black open histogram).
