Anti-Mouse OVA257-264 (SIINFEKL) peptide bound to H-2Kb PE
Also known as: H-2Kb-SIINFEKL, OVA-Kb
RUO: For Research Use Only
Staining of BALB/c (left) or C57BL/6 splenocytes (right) either unpulsed (open histogram) or pulsed with OVA257-264 (SIINFEKL) peptide (filled histogram) with 0.25 µg of Anti-Mouse OVA257-264 (SIINFEKL) peptide bound to H-2Kb PE. Total viable cells were used for analysis.
Contents: Anti-Mouse OVA257-264 (SIINFEKL) peptide bound to H-2Kb PE Catalog Number: 12-5743 Concentration: 0.2 mg/ml Formulation: Phosphate buffered saline, pH 7.2, less than or equal to 0.09% NaN3 Storage Conditions: Store at 2-8°C. Do not freeze. Light sensitive material. Clone: eBio25-D1.16 (25-D1.16) Host/Isotype: Mouse IgG1, κ
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12-5743-81
50 ug
12-5743-82
100 ug
Note: Several countries will continue to be supplied via distributors. Country specific prices may apply.
Flow Cytometry Product Notes: Test Sizes: To accommodate multicolor flow cytometry, eBioscience is in the process of reducing test size volumes from 20 µl to 5 µl. Please check your antibody vial for the recommended test size. Fluorochrome Replacements: eBioscience is in the process of replacing all Alexa Fluor® 647 conjugated products with eFluor® 660 conjugated products.
Description
The 25-D1.16 monoclonal antibody reacts with the 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. This antibody has proven to be very useful tracking the quantity and localization of these specific antigen-presenting cells (APC) in vivo.
Applications Reported
For research use only, not for diagnostic or therapeutic use. This eBio25-D1.16 (25-D1.16) antibody has been reported for use in flow cytometric analysis.
Applications Tested
This eBio25-D1.16 (25-D1.16) antibody has been tested by flow cytometric analysis of SIINFEKL-pulsed C57BL/6 splenocytes. This can be used at less than or equal to 0.5 μg per test. A test is defined as the amount (μg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 105 to 108 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.
Cells can be pulsed with the SIINFEKL peptide according to the following protocol:
1. With cells in flow staining buffer, add SIINFEKL peptide to a final concentration of 30 μM.
2. Incubate cells at 37°F for 2 hours.
3. Wash cells with flow staining buffer.
4. Proceed with cell surface staining as normal.
For additional information see the references listed below.
References
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 Jun;6(6):715-26. (25-D1.16, mAb development, PubMed)
Messaoudi I, LeMaoult J, Nikolic-Zugic J.The mode of ligand recognition by two peptide:MHC class I-specific monoclonal antibodies.J Immunol. 1999 Sep 15;163(6):3286-94.
Ackerman AL, Kyritsis C, Tampé R, Cresswell P.Access of soluble antigens to the endoplasmic reticulum can explain cross-presentation by dendritic cells.Nat Immunol. 2005 Jan;6(1):107-13.
Berwin B, Hart JP, Rice S, Gass C, Pizzo SV, Post SR, Nicchitta CV.Scavenger receptor-A mediates gp96/GRP94 and calreticulin internalization by antigen-presenting cells.EMBO J. 2003 Nov 17;22(22):6127-36.
