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Invitrogen™ IgM Monoclonal Antibody (SA-DA4), PerCP-eFluor™ 710, eBioscience™, Invitrogen™
Description
Description: The SA-DA4 monoclonal antibody reacts with the mu heavy chain of human IgM. It does not react with other classes of human immunoglobulin including IgD, IgG, or IgA. IgM is expressed intracellularly, during early stages of B lymphopoiesis, and then on the surface of mature B cells. Applications Reported: This SA-DA4 antibody has been reported for use in flow cytometric analysis. Applications Tested: This SA-DA4 antibody has been pre-titrated and tested by flow cytometric analysis of normal human peripheral blood cells. This can be used at 5 μL (0.06 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. PerCP-eFluor™ 710 emits at 710 nm and is excited with the blue laser (488 nm); it can be used in place of PerCP-Cyanine5.5. We recommend using a 710/50 bandpass filter, however, the 695/40 bandpass filter is an acceptable alternative. Please make sure that your instrument is capable of detecting this fluorochrome. Fixation: Samples can be stored in IC Fixation Buffer (cat. 00-8222) (100 μL cell sample + 100 μL IC Fixation Buffer) or 1-step Fix/Lyse Solution (cat. 00-5333) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation.
Specifications
Specifications
| Antigen | Human IgM |
| Applications | Flow Cytometry |
| Classification | Monoclonal |
| Clone | SA-DA4 |
| Concentration | 5 μL/Test |
| Conjugate | PerCP-eFluor 710 |
| Formulation | PBS with BSA and 0.09% sodium azide; pH 7.2 |
| Gene | IGHM |
| Gene Accession No. | P01871 |
| Gene Alias | AGM1; IGHM; immunoglobulin heavy constant mu; MU; VH |
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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 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.
For Research Use Only.
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