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Invitrogen™ Ly-6D Monoclonal Antibody (49-H4), eFluor™ 450, eBioscience™
Description
Description: This 49-H4 monoclonal antibody reacts with mouse Ly-6D, which is also known as ThB. This member of the Ly6 family of GPI-anchored cell surface glycoproteins is expressed on thymocytes and splenic B cells. Moreover, common lymphoid progenitor cells that express Ly-6D have been shown to commit to the B cell lineage. Expression of Ly-6D has also been associated with Rag-1 expression along with a reduced ability to differentiate into T or NK cells. Finally, reports indicate that the level of Ly-6D expression varies in different mouse strains. Applications Reported: This 49-H4 antibody has been reported for use in flow cytometric analysis. Applications Tested: This 49-H4 antibody has been tested by flow cytometric analysis of 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 10^5 to 10^8 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest. eFluor® 450 is an alternative to Pacific Blue®. eFluor® 450 emits at 445 nm and is excited with the Violet laser (405 nm). Please make sure that your instrument is capable of detecting this fluorochome. Excitation: 405 nm; Emission: 445 nm; Laser: Violet Laser. Filtration: 0.2 μm post-manufacturing filtered.
Specifications
Specifications
| Antigen | Ly-6D |
| Applications | Flow Cytometry |
| Classification | Monoclonal |
| Clone | 49-H4 |
| Concentration | 0.2 mg/mL |
| Conjugate | eFluor 450 |
| Formulation | PBS with 0.09% sodium azide; pH 7.2 |
| Gene | Ly6d |
| Gene Accession No. | P35459 |
| Gene Alias | E48; E48 antigen; Ly61; Ly-61; LY6D; Ly-6D; lymphocyte antigen 6 complex, locus D; lymphocyte antigen 6D; Thb; thymocyte B-cell antigen |
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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.
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