Anti-Human CXCR4/CD184 (12G5) 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.

C-X-C chemokine receptor type 4 (UniProt: P61073; also known as CXC-R4, CXCR-4, FB22, Fusin, HM89, LCR1, Leukocyte-derived seven transmembrane domain receptor, LESTR, Lipopolysaccharide-associated protein 3, LAP-3, LPS-associated protein 3, NPYRL, Stromal cell-derived factor 1 receptor, SDF-1 receptor, CD184) is encoded by the CXCR4 gene (Gene ID: 7852) in human. CXCR-4 is a multi-pass membrane protein with four extracellular domains, seven transmembrane domains, and four cytoplasmic domains. It serves as a receptor for the C-X-C chemokine CXCL12/SDF-1 and transduces signal by increasing intracellular calcium levels and enhancing MAPK1/MAPK3 activation. CXCR-4 is expressed in numerous tissues and two isoforms have been reported that are generated by alternative splicing. Isoform 1 is the predominant form in all tissues tested. CXCR-4 is rapidly phosphorylated on serine and threonine residues in the C-terminal region upon agonist stimulation. Phosphorylation at serine 324 and 325 leads to the recruitment of ITCH (E3 ubiquitin-protein ligase) that leads to its ubiquitination and degradation. Sulfation on tyrosine 21 is shown to be essential for efficient binding of CXCL12/SDF-1alpha and it promotes its dimerization. Tyrosine 7 and tyrosine 12 are sulfated in a sequential manner following complete sulfation of tyrosine 21. CXCR-4 acts as a co-receptor for human HIV-1 virus isolates (CD4 being the primary receptor) and as a primary receptor for HIV-2 isolates. CXCR-4 can also bind bacterial lipopolysaccharide (LPS) and mediate LPS-induced inflammatory response, including TNF-alpha secretion by monocytes. CXCR-4 is reported to be essential for metastatic spread to organs where CXCL12 is expressed, which allows tumor cells to access cellular niches that favor tumor-cell survival and growth. Mutations in CXCR4 gene are reported to cause WHIM syndrome that is characterized by neutropenia, hypogammaglobulinemia and extensive human papillomavirus (HPV) infection. (Ref.: Veldkamp, CT., et al. (2008). Sci. Signal. 1(37); ra4; Burger, JA., and Kipps, TJ (2006). Blood 107(5);1761-1767).

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Simian immunodeficiency virus (SIVmac) variant CP-MAC-infected Sup-T1 human T lymphoblasts.

Quality Control Testing

Evaluated by Flow Cytometry in Jurkat cells.

Flow Cytometry Analysis: Staining of one million Jurkat cells was performed using 5 μL of a 1:1 mixture of Cat. No. CWA-1125, Anti-Human CXCR4/CD184 (12G5) ColorWheelColorWheel^® Dye-Ready mAb and Cat. No. CWDS-PE ColorWheelColorWheel^® Antibody-Ready Phycoerythrin (PE) Dye or an equivalent amount of PE-conjugated Mouse IgG2a isotype control.

Tested Applications

Flow Cytometry Analysis: Staining of one million Jurkat cells was performed using 5 μL of a 1:1 mixture of Cat. No. CWA-1125, Anti-Human CXCR4/CD184 (12G5) ColorWheelColorWheel^® Dye-Ready mAb and Cat. No. CWDS-APC ColorWheelColorWheel^® Antibody-Ready Allophycocyanin (APC) Dye or an equivalent amount of APC-conjugated Mouse IgG2a 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 Jurkat cells.

Flow Cytometry Analysis: Staining of one million Jurkat cells was performed using 5 μL of a 1:1 mixture of Cat. No. CWA-1125, Anti-Human CXCR4/CD184 (12G5) ColorWheelColorWheel^® Dye-Ready mAb and Cat. No. CWDS-PE ColorWheelColorWheel^® Antibody-Ready Phycoerythrin (PE) Dye or an equivalent amount of PE-conjugated Mouse IgG2a isotype control.

Clone 12G5 is a mouse monoclonal antibody that detects human CXCR4 (CD184). It targets an epitope within the extracellular domain

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.