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Safety Data Sheet (SDS) APC Anti-mouse H-2Kb Bound To SIINFEKL Antibody     Product Data Sheet (PDF)    
APC Anti-mouse H-2Kb Bound To SIINFEKL Antibody
1308025 25 µg $90.00       
1308030 100 µg $290.00       
Clone: 25-D1.16
Isotype: Mouse IgG1, κ
Reactivity: Mouse
Immunogen: SIINFEKL pulsed RMA-S cells
Formulation: Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide.
Preparation: The antibody was purified by affinity chromatography and conjugated with APC under optimal conditions. The solution is free of unconjugated APC and unconjugated antibody.
Concentration: 0.2 mg/ml
Storage & Handling: The antibody solution should be stored undiluted between 2°C and 8°C, and protected from prolonged exposure to light. Do not freeze.
Application:

FC - Quality tested
IF - Validated

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.

Recommended Usage:

Each lot of this antibody is quality control tested by immunofluorescent staining with flow cytometric analysis. For flow cytometric 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.

Application References:

1. Mareeva T, et al. 2010. J. Immunol. Methods 353:78. (WB)
2. Dolan BP, et al. 2010. J. Immunol. 184:1419. (IF)
3. Porgador A, et al. 1997. Immunity 6:715. (WB, IF, IHC)

C57BL/6 mouse splenocytes were pulsed

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

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.



Description:

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.