Anti-Mouse OX40L/CD252 (RM134L) ColorWheel^® Dye-Ready mAb

ColorWheel^® technology is a novel and proprietary method of creating your own antibody and dye combinations for use in flow cytometry. By incubating any ColorWheel^® antibody with any ColorWheel^® dye, researchers can quickly and simply produce primary conjugated antibodies for use in single color or multicolor flow cytometry analysis. Each ColorWheel^® antibody and dye is lyophilized for long-term storage, allowing for simplicity and flexibility without compromise. ColorWheel^® technology requires both ColorWheel^® antibody and ColorWheel^® dye for Flow Cytometry application.

Tumor necrosis factor ligand superfamily member 4 (UniProt: P43488; also known as OX40 Ligand, OX40L, CD252) is encoded by the Tnfsf4 (also known as Ox40l, Txgp1l) gene (Gene ID: 22164) in murine species. OX40L, a member of the tumor necrosis factor family, is a single-pass type II membrane protein that contains a cytoplasmic domain (aa 1-28), a transmembrane domain (aa 29-50), and an extracellular domain (aa 51-198). It is a co-stimulatory molecule that binds to OX40 (TNFDF4) expressed on T cells, NK cells, B cells, and NKT cells. OX40L is broadly expressed by antigen presenting cells, activated T cells, and endothelial cells. OX40-OX40L interaction plays a key role in inducing T helper cells 2 (Th2) polarization and inhibiting T regulatory cell development. This interaction is also involved in many aspects of physiological responses between T cells and lymphoid and non-lymphoid cells. OX40/OX40L interactions is reported to play a key role in the development of multiple inflammatory and autoimmune diseases, making them attractive candidates for drug development. The absence of OX40/OX40L interactions is reported to suppress immune disease development in animal models. This interaction also plays a crucial role in the pathogenesis of experimental allergic encephalomyelitis (EAE). Mice deficient in OX40L exhibit poorly developed EAE, and OX40L transgenic mice display enhanced EAE symptoms. Blockade of OX40-OX40L interaction in arthritic mice is shown to reduce inflammation and restore tissue integrity predominantly by inhibiting inflammatory cytokine production by OX40L-expressing macrophages. (Ref.: Findlay, EG., et al. (2014). Proc. Natl. Acad. Sci. USA. 111(6); 2289-2294; Croft, M., et al. (2009). Immunol. Rev. 229(1); 173-191; Baum, PR., et al. (1994). EMBO J. 13(17); 3992-4001).

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Rat NRK-52E cells transfected with mouse OX-40L/CD252.

Quality Control Testing

Evaluated by Flow Cytometry in CHOZN cells stably transfected with mouse OX40L/CD252.

Flow Cytometry Analysis: Staining of one million CHOZN cells stably transfected with mouse OX40L/CD252 was performed using 5 µL of a 1:1 mixture of Cat. No. CWA-1160, Anti-Mouse OX40L/CD252 (RM134L) ColorWheel^® Dye-Ready mAb and Cat. No. CWDS-PE ColorWheel^® Antibody-Ready Phycoerythrin (PE) Dye or an equivalent amount of Cat. No. CWA-1012 Rat IgG2b isotype control and Cat. No. CWDS-PE ColorWheel^® Antibody-Ready Phycoerythrin (PE) Dye.

Tested Applications

Flow Cytometry Analysis: Staining of one million CHOZN cells stably transfected with mouse OX40L/CD252 was performed using 5 µL of a 1:1 mixture of Cat. No. CWA-1160, Anti-Mouse OX40L/CD252 (RM134L) ColorWheel^® Dye-Ready mAb and Cat. No. CWDS-PE ColorWheel^® Antibody-Ready Phycoerythrin (PE) Dye or an equivalent amount of Cat. No. CWA-1012 Rat IgG2b isotype control and Cat. No. CWDS-PE ColorWheel^® Antibody-Ready Phycoerythrin (PE) Dye.

Flow Cytometry Analysis: Staining of one million L5178Y cells stably transfected with mouse OX40L/CD252 was performed using 5 µL of a 1:1 mixture of Cat. No. CWA-1160, Anti-Mouse OX40L/CD252 (RM134L) ColorWheel® Dye-Ready mAb and Cat. No. CWDS-APC ColorWheel^® Antibody-Ready Allophycocyanin (APC) Dye or an equivalent amount of Cat. No. CWA-1012 Rat IgG2b isotype control and Cat. No. CWDS-APC ColorWheel® Antibody-Ready Allophycocyanin (APC) Dye.

Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user.

Clone RM134L is a rat monoclonal antibody that detects OX40L/CD252.

Lyophilized from PBS containing D-Mannitol and Sucrose, normal appearance is a dried pellet. Reconstituted antibody solution is stable and functional as assessed by functional testing. Contains no biocide or preservatives, such as azide.

1.0 mg/mL after reconstitution at 25 µL in PBS. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type.

Recommend storage of lyophilized product at 2-8°C. Before reconstitution, micro-centrifuge vials briefly to spin down material to bottom of the vial. Reconstitute each vial by adding 25 µL PBS. Protect from light.

ColorWheel is a registered trademark of Merck KGaA, Darmstadt, Germany

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.