MHC Class II I-Ab Monoclonal Antibody (AF6-120.1), Super Bright™ 780, eBioscience™
Isotype: IgG2a, kappa
Format: Super Bright™ 780
Application: Flow Cytometry
Tested Dilution: 1.0 µg/test
Concentration: 0.2 mg/mL
Storage: 4° C, store in dark, DO NOT FREEZE!
Formulation: PBS, pH 7.2, containing 0.09% sodium azide
Purification: Affinity chromatography
Data Sheet: TDS
Description: This AF6-120.1.2 monoclonal antibody reacts with the mouse MHC Class II I-Ab alloantigen of H-2b bearing mouse strains, including C57Bl/6 and 129. This cell surface molecule is involved in antigen presentation to T cells expressing CD3/TCR and CD4.
The AF6-120.1 antibody crossreacts with H-2k and H-2u haplotypes. However, reactivity to other haplotypes (i.e., d, f, g7, p, q, r, and s) has not been observed.
Applications Reported: This AF6-120.1.2 antibody has been reported for use in flow cytometric analysis.
Applications Tested: This AF6-120.1.2 antibody has been tested by flow cytometric analysis of mouse splenocytes. This may be used at less than or equal to 1.0 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 10^5 to 10^8 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.
Super Bright 780 is a tandem dye that can be excited with the violet laser line (405 nm) and emits at 780 nm. We recommend using a 780/60 bandpass filter. Please make sure that your instrument is capable of detecting this fluorochrome.
In some experiments, we have observed that compensation values for Super Bright 780-conjugated antibodies are higher in the violet 450/50 channel when using UltraComp eBeads microspheres (Product # 01-2222-42) as compared to single-color stained cells. In such circumstances, we would recommend setting compensation with cells. We have also observed this in some experiments using AbC Total Antibody Compensation beads (Product # A10497).
When using two or more Super Bright dye-conjugated antibodies in a staining panel, it is recommended to use Super Bright Complete Staining Buffer (Product # SB-4401) to minimize any non-specific polymer interactions. Please refer to the datasheet for Super Bright Staining Buffer for more information.
Light sensitivity: This tandem dye is sensitive to photo-induced oxidation. Please protect this vial and stained samples from light.
Fixation: Samples can be stored in IC Fixation Buffer (Product # 00-8222-49) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333-57) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically.
Excitation: 405 nm; Emission: 780 nm; Laser: Violet Laser
Super Bright Polymer Dyes are sold under license from Becton, Dickinson and Company.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.