ANTI-HUMAN E-SELECTIN/CD62E (HAE-1F) COLORWHEEL DYE-READY MAB

General description

ColorWheelColorWheel^® technology is a novel and proprietary method of creating your own antibody and dye combinations for use in flow cytometry. By incubating any ColorWheelColorWheel^® antibody with any ColorWheelColorWheel^® dye, researchers can quickly and simply produce primary conjugated antibodies for use in single color or multicolor flow cytometry analysis. Each ColorWheelColorWheel^® antibody and dye is lyophilized for long-term storage, allowing for simplicity and flexibility without compromise. ColorWheelColorWheel^® technology requires both ColorWheelColorWheel^® antibody and ColorWheelColorWheel^® 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 HAE-1f is a mouse monoclonal antibody that detects human E-Selectin (CD62E).

Immunogen

TNF-a activated HUVEC and human CD40 cDNA-transfected cells.

Application

Quality Control Testing

Evaluated by Flow Cytometry in TNF-a stimulated HUVECs.

Flow Cytometry Analysis: One million unstimulated HUVECs or TNF-a (10 ng/mL for 6 h)-stimulated HUVECs were stained with 5 μL of a 1:1 mixture of Cat. No. CWA-1120, Anti-Human E-Selectin/CD62E (HAE-1f) ColorWheelColorWheel^® Dye-Ready mAb and Cat. No. CWDS-PE ColorWheelColorWheel^® Antibody-Ready Phycoerythrin (PE) Dye or an equivalent amount of Cat. No. CWA-1013 Mouse IgG1 isotype control and Cat. No. CWD-PE ColorWheelColorWheel^® Antibody-Ready Phycoerythrin (PE) Dye.

Tested Applications

Flow Cytometry Analysis: One million unstimulated HUVECs or TNF-a (10 ng/mL for 6 h)-stimulated HUVECs were stained with 5 μL of a 1:1 mixture of Cat. No. CWA-1120, Anti-Human E-Selectin/CD62E (HAE-1f) ColorWheelColorWheel^® Dye-Ready mAb and Cat. No. CWDS-APC ColorWheelColorWheel^® Antibody-Ready Allophycocyanin (APC) Dye or an equivalent amount of Cat. No. CWA-1013 Mouse IgG1 isotype control and Cat. No. CWDS-APC ColorWheelColorWheel^® 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

Compatibility

Requires ColorWheel^® Dye (Sold Separately)

Target description

E-selectin (UniProt: P16581; also known as CD62 antigen-like family member E, Endothelial leukocyte adhesion molecule 1, ELAM-1, Leukocyte-endothelial cell adhesion molecule 2, LECAM2, CD62E) is encoded by the SELE (also known as ELAM1) gene (Gene ID: 6401) in human. Selectins (CD62s) are single-chain transmembrane glycoproteins that function as cell adhesion molecules (CAMs) by interacting with a range of oligosaccharide structures including sialyl-Lewis X (sLeX) via their C-type lectin domain. All three known selectins (E-, L-, and P-selectin) share a similar overall domain structure. E-selectin and P-selectin are mainly expressed on endothelial cells at inflammatory sites, whereas L-selectin is exclusively expressed on leukocytes. E-selectin expression on endothelial cells is transcriptionally upregulated by various proinflammatory substances such as IL-1, TNF and lipopolysaccharide (LPS). After endothelial cell stimulation, newly synthesized E-selectin is detected with a maximal surface expression after 3 to 6 hours and it returns to basal levels within 24 hours. It binds to sialofucosylated glycan determinants displayed on protein and lipid scaffolds of blood cells. E-selectin binding determinants are typically displayed at the end of O-glycans, N-glycans, or glycolipid precursor structures, and require the coordinated and sequential action of specific glycosyltransferases localized within the lumen of the Golgi apparatus. E-Selectin is a single-pass type I membrane protein that is synthesized with a signal peptide (aa 1-21), which is subsequently cleaved off to produce the mature form that contains an extracellular domain (aa 22-556), a transmembrane domain (aa 557-578), and a cytoplasmic domain (aa 579-610). At its N-terminal it has a C-type lectin domain (aa 22-139) and an EGF-like domain (aa 140-175). It also has 6 Sushi domains. (Ref.: Silva, M., et al. (2018). Front. Immunol. 8; 1878).

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.