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Invitrogen™ SIGLEC H Monoclonal Antibody (eBio440c), PerCP-eFluor™ 710, eBioscience™
Description
The monoclonal antibody eBio440c recognizes Siglec-H, a protein exclusively found on pDC (plasmacytoid dendritic cells) or type I IFN-producing cells (IPC) in the naive mouse. mouse IPC are typically PDCA+, CD11c+, CD11b-, B220+, and Ly-6C+, and are quick to respond to viruses. Siglec-H is a transmembrane protein of the Ig superfamily that like CD33 have been shown to bind sialic acid but lacks the characteristic cytoplasmic ITIM domain (immunoreceptor tyrosine based inhibitory motif). To overcome the lack of a cytoplasmic domain, Siglec-H associates with DAP12 thereby allowing for signal transduction. The eBio440c antibody has been shown to inhibit pDC function (inhbits IFNalpha secretion in response to CpG). It has been observed that some mouse strains (such as SJL) have higher percentages of pDCs compared to C57BL/6. Applications Reported: This eBio440c antibody has been reported for use in flow cytometric analysis. Applications Tested: This eBio440c antibody has been tested by flow cytometric analysis of SJL spleen cells. 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. PerCP-eFluor 710 emits at 710 nm and is excited with the blue...
Specifications
Specifications
| Antigen | SIGLEC H |
| Applications | Flow Cytometry |
| Classification | Monoclonal |
| Clone | eBio440c |
| Concentration | 0.2 mg/mL |
| Conjugate | PerCP-eFluor 710 |
| Formulation | PBS with 0.09% sodium azide; pH 7.2 |
| Gene | Siglech |
| Gene Accession No. | 0 |
| Gene Alias | 6430529G09Rik; sialic acid binding Ig-like lectin H; Siglec; Siglech; Siglec-H; SIGLEC-like protein |
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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.
As with other fluorochromes, we recommend minimal exposure to light to maintain optimal signal.
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