ANTI-HUMAN NESTIN (8B8) 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. This ColorWheel^® Dye-Ready monoclonal antibody was developed using our proprietary ZooMAb^® recombinant antibody technology. It offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals.

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 8B8 is a recombinant rabbit monoclonal antibody that detects Human Nestin. It targets an epitope within the C-terminal region.

Immunogen

GST-tagged recombinant fragment corresponding to 150 amino acid from the C-terminal region of human Nestin.

Application

Quality Control Testing

Evaluated by Flow Cytometry in Fixed and permeabilized SH-SY5Y cells.

Flow Cytometry Analysis: Intracellular staining of one million fixed and permeabilized SH-SY5Y cells was performed using 5 μL of a 1:1 mixture of Cat. No. CWA-1132, Anti-Human Nestin (8B8) ColorWheelColorWheel^® Dye-Ready mAb and Cat. No. CWDS-PE ColorWheelColorWheel^® Antibody-Ready Phycoerythrin (PE) Dye or an equivalent amount of PE-conjugated Rabbit IgG isotype control.

Tested Applications

Flow Cytometry Analysis: Intracellular staining of one million fixed and permeabilized SH-SY5Y cells was performed using 5 μL of a 1:1 mixture of Cat. No. CWA-1132, Anti-Human Nestin (8B8) ColorWheelColorWheel^® Dye-Ready mAb and Cat. No. CWDS-APC ColorWheelColorWheel^® Antibody-Ready Allophycocyanin (APC) Dye or an equivalent amount of APC-conjugated Rabbit IgG 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 Fixed and permeabilized SH-SY5Y cells.

Flow Cytometry Analysis: Intracellular staining of one million fixed and permeabilized SH-SY5Y cells was performed using 5 μL of a 1:1 mixture of Cat. No. CWA-1132, Anti-Human Nestin (8B8) ColorWheelColorWheel^® Dye-Ready mAb and Cat. No. CWDS-PE ColorWheelColorWheel^® Antibody-Ready Phycoerythrin (PE) Dye or an equivalent amount of PE-conjugated Rabbit IgG isotype control.

Compatibility

Requires ColorWheel^® Dye (Sold Separately)

Target description

Nestin (UniProt: P48681) is encoded by the NES (also known as Nbla00170) gene (Gene ID: 10763) in human. Nestin is a class VI intermediate filament protein that is required for survival, renewal, and mitogen-stimulated proliferation of neural progenitor cells. Nestin expression is upregulated in most mitotic cells and is downregulated upon differentiation. Upon terminal neural differentiation Nestin levels are shown to be down regulated and replaced by neurofilaments. Nestin is also expressed in kidney in a developmentally regulated pattern. Its expression is associated with tubulointerstitial injury and can serve as a marker for peritubular endothelial cell injury and tubulointerstitial fibrosis. Nestin is reported to be constitutively phosphorylated, and its phosphorylation state increases during mitosis when the cytoplasmic intermediate filament network is reorganized. Nestin has a short N-terminus and an unusually long C-terminus region that results in complex binding to an array of structural proteins. The short N-terminus leads to preferential intermediate filaments formation with heterodimers and heterotetramers and the long C-terminal portion functions as links or bridges between intermediate filaments and microtubules. Nestin can exist in homodimeric or homotetrameric form, however in mixtures with other intermediate filament proteins such as vimentin and alpha-internexin, it can form heterodimers, which can assemble to form intermediate filaments. Nestin is shown to promote the disassembly of phosphorylated vimentin intermediate filaments during mitosis and play a role in the trafficking and distribution of intermediate filament proteins to daughter cells during progenitor cell division. (Ref.: Xie, L., et al. (2015). Stem Cells Int. 2015:762098; Tomioka, M., et al. (2010). 15(5); 568-574).This ColorWheel^® Dye-Ready monoclonal antibody was developed using our ZooMAb^® recombinant monoclonal antibody (Cat. No. ZRB69). It offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals.

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

ZooMAb 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.