ANTI-MOUSE PD-L1/CD274 (10F.9G2) COLORWHEEL DYE-READY MAB

General description

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.

We are committed to bringing you greener alternative products, which adhere to one or more of The 12 Principles of Green Chemistry. This product is Preservative-free, lyophilized product for enhanced stability and allow for ambient shipping and thus aligns with "Waste Prevention", "Designing Safer Chemicals" and "Design for Energy Efficiency". Click here for more information.

Specificity

Clone 10F.9G2 is a rat monoclonal antibody that detects murine PD-L1 (CD274). It targets an epitope within the extracellular domain.

Immunogen

Chinese Hamster ovary (CHO) cells transfected with murine PD-L1/CD274.

Application

Quality Control Testing

Evaluated by Flow Cytometry in LPS-stimulated RAW264.7 cells.

Flow Cytometry Analysis: One million RAW264.7 cells either unstimulated or stimulated with lipopolysaccharide (LPS; 1 μg/mL 16 h) were stained with 5 μL of a 1:1 mixture of Cat. No. CWA-1131, Anti-Mouse PD-L1/CD274 (10F.9G2) ColorWheelColorWheel^® Dye-Ready mAb and Cat. No. CWDS-PE ColorWheelColorWheel^® Antibody-Ready Phycoerythrin (PE) Dye or an equivalent amount of PE-conjugated Rat IgG2b isotype control.

Tested Applications

Flow Cytometry Analysis: One million RAW264.7 cells either unstimulated or stimulated with Lipopolysaccharide (LPS; 1 μg/mL; 16 h) were stained with 5 μL of a 1:1 mixture of Cat. No. CWA-1131, Anti-Mouse PD-L1/CD274 (10F.9G2) ColorWheelColorWheel^® Dye-Ready mAb and Cat. No. CWDS-APC ColorWheelColorWheel^® Antibody-Ready Allophycocyanin (APC) Dye or an equivalent amount of APC-conjugated Rat IgG2b isotype control.

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

Evaluated by Flow Cytometry in LPS-stimulated RAW264.7 cells.

Flow Cytometry Analysis: One million RAW264.7 cells either unstimulated or stimulated with lipopolysaccharide (LPS; 1 μg/mL 16 h) were stained with 5 μL of a 1:1 mixture of Cat. No. CWA-1131, Anti-Mouse PD-L1/CD274 (10F.9G2) ColorWheelColorWheel^® Dye-Ready mAb and Cat. No. CWDS-PE ColorWheelColorWheel^® Antibody-Ready Phycoerythrin (PE) Dye or an equivalent amount of PE-conjugated Rat IgG2b isotype control.

Compatibility

Requires ColorWheel^® Dye (Sold Separately)

Target description

Programmed cell death 1 ligand 1 (UniProt: Q9EP73; also known as PD-L1, PDCD1 ligand 1, Programmed death ligand 1, B7 homolog 1, B7-H1, CD274) is encoded by the Cd274 (also known as B7h1, Pdcd1l1, Pdcd1lg1, Pdl1) gene (Gene ID: 50533) in murine species. PD-L1 is a single-pass type I membrane glycoprotein that is synthesized with a signal peptide (aa 1-18), which is subsequently cleaved off to produce the mature form that contains a large extracellular domain (aa 19-239) with an Ig-like V type domain (aa 19-127) and an Ig-like C2 -type domain (aa 133-224), followed by a transmembrane domain (aa 240-260), and a cytoplasmic tail (aa 261-290). It is highly expressed in the heart, thymus, skeletal muscle, and lung. Its expression is also reported on activated dendritic cells, B-cells, and macrophages. It serves as an immune checkpoint inhibitor that binds to its receptor PD-1 expressed by T cells and other immune cells to regulate immune responses. Many tumor cells exploit this mechanism by overexpressing PD-L1 which often correlates with poor prognosis. PD-L1 expression can be up-regulated by interferon- treatment in monocytes, B cells, and macrophages. PD-1 and its ligands, PD-L1 and PD-L2 play a key role in the maintenance of peripheral tolerance, a process by which the quiescence of autoreactive mature T cells is maintained. However, tumors and pathogens that cause chronic infections can exploit this pathway to escape T-cell mediated tumor-specific and pathogen-specific immunity. The interaction with PD-1/PD-L1 inhibits cytotoxic T lymphocytes effector function. The effector functions of T-cells expressing PD-1 can be downregulated by PD-L1 or PD-L2 expressed by the tumor cells. The blockage of the PD-1/PD-L1-mediated pathway results in the reversal of the exhausted T-cell phenotype and the normalization of the anti-tumor response, providing a rationale for cancer immunotherapy. (Ref.: Hudson, K., et al. (2020). Front. Immunol. 11; 568931; Iwai, Y., et al. (2002). Proc. Natl. Acad. Sci. USA. 99)19); 12293-12297).

Physical form

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.

Reconstitution

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.

Storage and Stability

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.

Legal Information

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

Disclaimer

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.