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Invitrogen™ CD154 (CD40 Ligand) Monoclonal Antibody (MR1), eFluor™ 450, eBioscience™
Description
Description: The MR1 monoclonal antibody reacts with mouse CD154, a 39 kDa transmembrane glycoprotein also known as CD40 ligand (CD40L). CD154 is expressed transiently by activated T cells. Through its binding to CD40 on antigen presenting cells (APC) including B cells, monocytes/macrophages, and dendritic cells, it serves a crucial function in T cell-APC cognate interaction. CD154-interaction with CD40 transduces signals for T-dependent B cell activation and induces B cells to enter the cell cycle. For staining for flow cytometric analysis, it is important to stimulate enriched T cells or enriched CD4 cells (using depletion strategy) prior to staining with MR1. Applications Reported: This MR1 antibody has been reported for use in flow cytometric analysis. Applications Tested: This MR1 antibody has been tested by flow cytometric analysis of stimulated mouse splenocytes. This can be used at less than or equal to 0.5 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; Emissi...
Specifications
Specifications
| Antigen | CD154 (CD40 Ligand) |
| Applications | Flow Cytometry |
| Classification | Monoclonal |
| Clone | MR1 |
| Concentration | 0.2 mg/mL |
| Conjugate | eFluor 450 |
| Formulation | PBS with 0.09% sodium azide; pH 7.2 |
| Gene | CD40LG |
| Gene Accession No. | P27548 |
| Gene Alias | CD154; CD40 antigen ligand; CD40 ligand; CD40 ligand CD154; CD40 ligand, membrane form; CD40 ligand, soluble form; CD40L; CD40-L; Cd40lg; gp39; hCD40L; H-CD-40-L; HIGM1; IGM; IMD3; Ly62; Ly-62; M-CD-40-L; RP23-153G22.3; T-B cell-activating molecule; T-BAM; T-cell antigen Gp39; TNF-related activation protein; TNFSF5; TRAP; tumor necrosis factor (ligand) superfamily member 5; tumor necrosis factor (ligand) superfamily, member 5; tumor necrosis factor ligand superfamily member 5 |
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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.
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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