ANTI-MOUSE C-KIT/CD117 (ACK2) 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 ACK2 is a rat monoclonal antibody that detects murine c-Kit/CD117. It targets an epitope within the extracellular domain.

Immunogen

IL-3 dependent mast cells derived from mouse bone marrow.

Application

Quality Control Testing

Evaluated by Flow Cytometry in Mouse Bone Marrow Cells.

Flow Cytometry Analysis: Staining of one million C57BL/6 mouse bone marrow cells was performed with APC-conjugated anti-mouse CD45R and with 5 μL of a 1:1 mixture of Cat. No. CWA-1133, Anti-Mouse c-Kit/CD117 (ACK2) ColorWheelColorWheel^® Dye-Ready mAb and Cat. No. CWDS-PE ColorWheelColorWheel^® Antibody-Ready Phycoerythrin (PE) Dye (right figure) or an equivalent amount of PE-conjugated Rat IgG2b isotype control. More CD117 expressing cells were observed in CD45R- population than in CD45R+ population.

Tested Applications

Flow Cytometry Analysis: Staining of one million C57BL/6 mouse bone marrow cells was performed with PE-conjugated anti-mouse CD45R and with 5 μL of a 1:1 mixture of Cat. No. CWA-1133, Anti-Mouse c-Kit/CD117 (ACK2) ColorWheelColorWheel^® Dye-Ready mAb and Cat. No. CWDS-APC ColorWheelColorWheel^® Antibody-Ready Allophycocyanin (APC) Dye (right figure) or an equivalent amount of APC-conjugated Rat IgG2b isotype control. More c-Kit/CD117 expressing cells were observed in CD45R- population than in CD45R+ population.

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 Mouse Bone Marrow Cells.

Flow Cytometry Analysis: Staining of one million C57BL/6 mouse bone marrow cells was performed with APC-conjugated anti-mouse CD45R and with 5 μL of a 1:1 mixture of Cat. No. CWA-1133, Anti-Mouse c-Kit/CD117 (ACK2) ColorWheelColorWheel^® Dye-Ready mAb and Cat. No. CWDS-PE ColorWheelColorWheel^® Antibody-Ready Phycoerythrin (PE) Dye (right figure) or an equivalent amount of PE-conjugated Rat IgG2b isotype control. More CD117 expressing cells were observed in CD45R- population than in CD45R+ population.

Compatibility

Requires ColorWheel^® Dye (Sold Separately)

Target description

Mast/stem cell growth factor receptor Kit (UniProt: P05532; also known as EC:2.7.10.1, SCFR, Proto-oncogene c-Kit, Tyrosine-protein kinase Kit, CD117) is encoded by the Kit (also known as SI) gene (Gene ID: 16590) in murine species. c-Kit is a single-pass type I membrane glycoprotein that is synthesized with a signal peptide (aa 1-24), which is subsequently cleaved off to generate the mature form that contains an extracellular domain (aa 25-527), a transmembrane domain (aa 528-548), and a cytoplasmic domain (aa 549-979). c-Kit has tyrosine protein kinase activity and serves as a cell-surface receptor for the stem cell factor (SCF)/KITLG. It plays a significant role in the regulation of cell survival, cell proliferation, stem cell maintenance, gametogenesis and other functions. In response to SCF binding, c-Kit can activate several signaling pathways. In the absence of bound SCF, it exists as a monomer, however, it homodimerizes in the presence of bound SCF and forms a heterotetramer with two SCF molecules. c-Kit can be autophosphorylated on tyrosine residues and SCF binding enhances its autophosphorylation. Its kinase activity is reported to be down-regulated by phosphorylation on serine residues by protein kinase C family members. Phosphorylation at Tyr 571 is shown to be essential for its interaction with PTPN11/SHP-2 and phosphorylation of Tyr 573 is required for interaction with PTPN6/SHP-1. c-Kit can be ubiquitinated by SOCS6 and it is rapidly ubiquitinated after autophosphorylation induced by SCF binding, leading to its internalization and degradation. Lack of c-Kit expression has been reported in multiple cancer types and c-Kit expressing cells display a regression of typical morphological features of malignancy. (Ref.: Foster, BM., et al. (2018). Biomedicines. 6(1); 31; Yee, NS., et al. (1994). J. Biol. Chem. 269(50); 1991-1998).

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.