Product Information

Contents: Affinity Purified anti-mouse CD154 (CD40 Ligand, CD40L, gp39)
Catalog Number: 14-1541
Sizes: 50 ug, 100 ug, 500 ug
Formulation: Phosphate buffer pH 7.2,
150 mM NaCl, 0.09% NaN₃
Storage Conditions: Store at 4°C.
Avoid repeated freeze/thaw cycles.
Clone: MR1
Isotype: Armenian Hamster IgG

Surface staining of PMA + Ionomycin stimulated isolated T cells from mouse splenocytes with anti-mouse CD154 (MR1) PE. Appropriate isotype controls were used (open histogram). Total viable cells were used for analysis.

Available Formats of This Product
Cat. No.	Format	Excite (nm)	Emit (nm)	Reported Applications
14-1541	Affinity Purified anti-mouse CD154 (CD40 Ligand, CD40L, gp39)	N/A	N/A	FA FC IHC
16-1541	Functional Grade* Purified anti-mouse CD154 (CD40 Ligand, CD40L, gp39)	N/A	N/A	FA FC
13-1541	Biotin anti-mouse CD154 (CD40 Ligand, CD40L, gp39)	N/A	N/A	FC
12-1541	PE anti-mouse CD154 (CD40 Ligand, CD40L, gp39)	488	575	FC
17-1541	Allophycocyanin (APC) anti-mouse CD154 (CD40 Ligand, CD40L, gp39)	633	660	FC
*Functional Grade™ (FG™) Purified: Azide-free, sterile-filtered, and endotoxin < 0.001 ng/µg (unless otherwise noted). *Functional Grade™ (FG™) Biotin: Azide-free, sterile-filtered, and endotoxin < 0.05 ng/µg. Purified: Contains azide, not sterile-filtered, and not endotoxin tested.
Flow Cytometry Product Notes: Test Sizes: To accommodate multicolor flow cytometry, eBioscience is in the process of reducing test size volumes from 20 µl to 5 µl. Please check your antibody vial for the recommended test size. Fluorochrome Replacements: eBioscience is in the process of replacing all Pacific Blue® and APC-Alexa Fluor® 750 conjugated products with eFluor™ 450 and APC-eFluor™ 780 conjugated products, respectively.

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Description

The MR1 monoclonal antibody reacts with mouse CD154, a 39 kDa transmembrane glycoprotein also known as gp39 and CD40 ligand (CD40L). gp39 is expressed transiently by activated T cells and through its binding to CD40 on antigen presenting cells including B cells, monocytes/macrophages and dendritic cells, serves a crucial function in T-APC cognate interaction. gp39 interaction with CD40 transduces signals for T-dependent B cell activation and induces B cell cycle entry.

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

For research use only, not for diagnostic or therapeutic use. The MR1 antibody has been reported for use in flow cytometric analysis, and immunohistochemical staining. It has also been reported to inhibit binding of CD154 to CD40 and activation of B cells. (Please use Functional Grade purified MR1, cat. 16-1541, in functional assays.)

Applications Tested

The MR1 antibody has been tested by flow cytometric analysis of resting and 6-8 hour activated splenocyte suspensions. This can be used at less than or equal to 0.5 μg per million cells in a 100 μl total staining volume. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.

References

Lesley R, Kelly LM, Xu Y, Cyster JG. Naive CD4 T cells constitutively express CD40L and augment autoreactive B cell survival. Proc Natl Acad Sci U S A. 2006 Jul 11;103(28):10717-22.

Roy M, Aruffo A, Ledbetter J, Linsley P, Kehry M, Noelle R. Studies on the interdependence of gp39 and B7 expression and function during antigen-specific immune responses. Eur J Immunol. 1995 Feb;25(2):596-603.

Yellin MJ, Sippel K, Inghirami G, Covey LR, Lee JJ, Sinning J, Clark EA, Chess L, Lederman S. CD40 molecules induce down-modulation and endocytosis of T cell surface T cell-B cell activating molecule/CD40-L. Potential role in regulating helper effector function. J Immunol. 1994 Jan 15;152(2):598-608.

Van den Eertwegh AJ, Noelle RJ, Roy M, Shepherd DM, Aruffo A, Ledbetter JA, Boersma WJ, Claassen E. In vivo CD40-gp39 interactions are essential for thymus-dependent humoral immunity. I. In vivo expression of CD40 ligand, cytokines, and antibody production delineates sites of cognate T-B cell interactions. J Exp Med. 1993 Nov 1;178(5):1555-65.

Roy M, Waldschmidt T, Aruffo A, Ledbetter JA, Noelle RJ. The regulation of the expression of gp39, the CD40 ligand, on normal and cloned CD4+ T cells. J Immunol. 1993 Sep 1;151(5):2497-510.

Noelle RJ, Roy M, Shepherd DM, Stamenkovic I, Ledbetter JA, Aruffo A. A 39-kDa protein on activated helper T cells binds CD40 and transduces the signal for cognate activation of B cells. Proc Natl Acad Sci U S A. 1992 Jul 15;89(14):6550-4.

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