Anti-Human OCT4 (4N8) 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.

POU domain, class 5, transcription factor 1 (UniProt: Q01860 also known as Octamer-binding protein 3, Oct-3, Octamer-binding protein 4, Oct-4, Octamer-binding transcription factor 3, OTF-3) is encoded by the POU5F1 (also known as OCT3, OCT4, and OTF3) (Gene ID: 5460) in human. OCT-4 is a transcription factor that binds to the octamer motif (5′-ATTTGCAT-3′). It forms a trimeric complex with SOX2 on DNA and controls the expression of several genes involved in embryonic development such as YES1, FGF4, UTF1 and ZFP206. It is shown to be critical for early embryogenesis and for embryonic stem cell pluripotency. It is highly expressed in undifferentiated embryonic stem cells and its expression decreases gradually after embryoid body (EB) formation. In humans, two isoforms of OCT-4 have been described that are generated by alternative splicing. The isoforms, OCT4-IA (360 amino acids) and OCT4-IB (265 amino acids) have identical C-terminal (225 amino acids) but differ in sequence at their N- terminal region. It has been reported that OCT4-IA is essential to sustain stem cell self-renewal and OCT4-IB is not related to stemness. Oct-4 contains a POU-specific domain (aa 138-212) that mediates its interaction with pyruvate kinase isozyme type M2 (PKM2), which regulates its transcriptional activity in as positive manner. Human OCT-4 can undergo phosphorylation by protein kinase A and/or ERK at a highly conserved serine 236 within the POU DNA-binding homeodomain. Phosphorylation at this site sterically hinders both its DNA binding and homodimer assembly. OCT-4 can undergo sumoylation that enhances its stability, DNA binding and transactivation activity. WWP2, an E3 ubiquitin ligase is shown to specifically interact with OCT-4 through its tryptophan-based WW domains, which promotes its ubiquitination and proteasomal degradation. (Ref.: Zeineddine, D., et al. (2014). Am. J. Stem Cells. 3(2);74-82; Lee, J., et al. (2008). Int. J. Biochem. Cell Biol. 40(5); 1043-54).
This ColorWheel^® Dye-Ready monoclonal antibody was developed using our ZooMAb^® recombinant monoclonal antibody (Cat. No. ZRB1101). 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.

His-tagged recombinant fragment corresponding to the first 134 amino acids from human Octamer-binding protein 4 (OCT4)

Quality Control Testing

Evaluated by Flow Cytometry in Fixed and permeabilized NTERA2 cells.

Flow Cytometry Analysis: Intracellular staining of one million fixed and permeabilized NTERA2 cells was performed using 5 μL of a 1:1 mixture of Cat. No. CWA-1124, Anti-Human OCT4 (4N8) 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 NTERA2 cells was performed using 5 μL of a 1:1 mixture of Cat. No. CWA-1124, Anti-Human OCT4 (4N8) 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 NTERA2 cells.

Flow Cytometry Analysis: Intracellular staining of one million fixed and permeabilized NTERA2 cells was performed using 5 μL of a 1:1 mixture of Cat. No. CWA-1124, Anti-Human OCT4 (4N8) 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.

Clone 4N8 is a recombinant rabbit monoclonal antibody that detects OCT4. It targets an epitope within the N-terminal region.

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

ZooMAb 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.