ANTI-MOUSE CD45.1 (A20) 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 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 A20 is a mouse monoclonal antibody that detects CD45.1 allotype in murine species.

Immunogen

SJL mouse thymocytes and splenocytes

Application

Quality Control Testing:

Evaluated by Flow Cytometry in Mouse splenocytes containing the 45.1 gene.

Staining of one million mouse splenocytes containing the 45.1 gene was performed using 5 μL of a 1:1 mixture of Cat. No. CWA-1058, Anti-Mouse CD45.1 (A20) ColorWheel^® Dye-Ready mAb and Cat. No. CWD-PE ColorWheel^® Antibody-Ready Phycoerythrin (PE) Dye or an equivalent amount of PE-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

Compatibility

Requires ColorWheel^® Dye (Sold Separately)

Target description

Receptor-type tyrosine-protein phosphatase C (UniProt: P06800; also known as EC:3.1.3.48, Leukocyte common antigen, L-CA, Lymphocyte antigen 5, Ly-5,T200, CD45) is encoded by the Ptprc (also known as Ly-5) gene (Gene ID: 19264) in murine species. CD45 is a single-pass type I membrane glycoprotein that is synthesized with a signal peptide (aa 1-25), which is subsequently cleaved off to generate the mature form that contains an extracellular domain (aa 26-566), a transmembrane domain (aa 567-588), and a cytoplasmic domain (aa 589-1293). It serves as a receptor-tyrosine phosphatase that is required for antigen-induced signaling in T-cells and B-cells. It contains two protein tyrosine phosphatase (PTPase) domains (aa 642-901 and 933-1216) and the first PTPase domain has enzymatic activity, while the second one seems to affect the substrate specificity of the first one. Its substrate binding site is localized to amino acids 842-848. CD45 functions as a signaling gatekeeper in T-cells that require CD45 to detect and respond to antigen because it dephosphorylates the C-terminal inhibitory tyrosines thereby activates Src family kinase Lck that phosphorylates the T cell antigen receptor (TCR). Upon T-cell activation, CD45 also it recruits and dephosphorylates SKAP1 and Fyn. Furthermore, it also dephosphorylated Lyn and modulates its activity. Clone A20 is typically used as a leukocyte marker in SJL/J, DA, STS/A and RIII mice. A single amino acid K302E determines if this will be CD45.1 or CD45.2. This antibody has been demonstrated to specific for CD45.1 and is not cross-reactive with CD45.2 bearing cells. (Ref.: Courtney, AH., et al. (2019). Sci. Sig. 12(604); eaaw8151; Mercier, FE., et al. (2016). Stem Cell Reports. 6(6); 985-992; Coughlin, S., et al. (2015). Proc. Natl. Acad. Sci. USA. 112(47); E6515-E6524).

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 of 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.