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Invitrogen™ OVA257-264 (SIINFEKL) peptide bound to H-2Kb Monoclonal Antibody (eBio25-D1.16 (25-D1.16)), PerCP-eFluor™ 710, eBioscience™
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: 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 peptide-pulsed mouse splenocytes. This can be used at less than or equal to 0.125 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 10e5 to 10e8 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest. PerCP-eFluor 710 can be used in place of PE-Cy5, PE-Cy5.5 or PerCP-Cy5.5. PerCP-eFluor 710 emits at 710 nm and is excited with the blue laser (488 nm). Please make sure that your instrument is capable of detecting this fluorochrome. For a filter configuration, we recommend using the 685 LP dichroic mirror and 710/40 band pass filter, however the 695/40 band pass filter is an acceptable alternative. Our testing indicates that PerCP-eFluor 710 conjugated antibodies are stable when stained samples are...
Specifications
Specifications
| Antigen | OVA257-264 (SIINFEKL) peptide bound to H-2Kb |
| Applications | Flow Cytometry |
| Classification | Monoclonal |
| Clone | eBio25-D1.16 (25-D1.16) |
| Concentration | 0.2 mg/mL |
| Conjugate | PerCP-eFluor 710 |
| Formulation | PBS with 0.09% sodium azide; pH 7.2 |
| Gene Alias | H-2Kb-SIINFEKL; OVA257-264; OVA257-264 H-2Kb; OVA-Kb |
| Host Species | Mouse |
| Purification Method | Affinity chromatography |
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Frequently Asked Questions (FAQs)
Our options will depend on the samples you are analyzing.
If cell viability is not critical, you can store your stained samples at 4 degrees C or on ice overnight in the dark and analyze the following day.
For samples stained with eFluor organic fluorochromes, we recommend that cells be suspended in 100 uL of Flow Cytometry Staining Buffer (Cat. No. 00-4222) and 100 uL of eBioscience IC Fixation Buffer (Cat. No. 00-8222); samples can be incubated for up to 3 days at 4 degrees C in the dark. Alternatively, the 1-step Fix/Lyse Solution (Cat. No. 00-5333) can be used. This is a great option when working with whole blood but also works for other cell types.
Yes, the eFluor Organic fluorochromes can be used for intracellular staining. The eFluor organic fluorochromes maintain bright signal and require minimal changes in compensation when fixed with eBioscience IC Fixation Buffer (Cat. No. 00-8222-49) and Permeabilization Buffer (Cat. No. 00-8333-56) or 1-step Fix/Lyse Solution (Cat. No. 00-5333-54, 00-5333-57) (as compared to live cells).
The eFluor Organic fluorochromes and eVolve QDots can be used with flow staining buffers containing PBS and protein.
The eFluor Organic Dyes (eFluor 450, APC-eFluor 780, PerCP-eFluor 710, eFluor 710) are conventional fluorochromes. In contrast, the eVolve line of products are Quantum dots.
As with other fluorochromes, we recommend minimal exposure to light to maintain optimal signal.
Safety and Handling
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