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Live Cell Imaging

Live-cell imaging allows the visualization of cellular processes using time-lapse microscopy. Structural changes and physiological processes can be observed in real-time. Other common microscopy techniques such as immunofluorescence usually require cell fixation and permeabilization, which shows only a snapshot of the cells at a certain timepoint and might lead to artefacts. In contrast, during live-cell imaging, dynamic changes can be analyzed, particularly, when subcellular structures are being investigated.

Chromobodies

Chromobodies are fluorescent nanoprobes for real-time, live-cell imaging of endogenous proteins. These Nanobodies, fused to a fluorescent protein (GFP or RFP), are expressed inside the cell and can bind to target proteins, marking them with fluorescence. They are compatible with various eukaryotic cells, from plants to vertebrates. Chromobodies can be introduced via transient transfection or used in stable cell lines and transgenic organisms with constitutive or responsive expression.

  • Non-invasive: Intracellular expression
  • Compatible with many host species

Immuno-Oncology VHHs New

Our Immuno-Oncology VHHs are monovalent Nanobodies that are targeted against the most important immune oncology targets like PD-1, PD-L1, TIGIT etc.. They are optimal tools for IF, SRM, Flow, but due to their small size also live cell imaging. Their monovalent nature helps avoiding potential experimental bias through clustering effects. A minimal epitope-label displacement is another important benefit when working with Nanobodies, resulting in sharper images with less diffuse fluorescent signal.

  • Minimal epitope label displacement
  • No receptor crosslinking with monovalent binding

Target Conjugates Applications
CTLA4 CoraLite® Plus 555 CoraLite® Plus 647 FITC IF, Live Cell Imaging, FC
FLT3 CoraLite® Plus 555 CoraLite® Plus 647 FITC IF, Live Cell Imaging, FC
LAG3 CoraLite® Plus 555 CoraLite® Plus 647 FITC IF, Live Cell Imaging, FC
MSLN CoraLite® Plus 555 CoraLite® Plus 647 FITC IF, Live Cell Imaging, FC
PD-1 CoraLite® Plus 555 CoraLite® Plus 647 FITC FC
PD-L1 CoraLite® Plus 555 CoraLite® Plus 647 FITC IF, Live Cell Imaging, FC
TIM3 CoraLite® Plus 555 CoraLite® Plus 647 FITC IF, Live Cell Imaging, FC
TIGIT CoraLite® Plus 555 CoraLite® Plus 647 FITC IF, Live Cell Imaging, FC

Target Format
Actin TagGFP2 TagRFP Sold in plasmid format; MTA required
Cell Cycle Chromobody TagRFP Sold in plasmid format; MTA required
DNMT1 Chromobody TagGFP2 TagRFP Sold in plasmid format; MTA required
Histone Chromobody EGFP Sold in plasmid format; MTA required
Lamin Chromobody TagGFP2 Sold in plasmid format; MTA required
mNeonGreen Chromobody mCherry Sold in plasmid format; MTA required
Nuclear Actin TagGFP Sold in plasmid format; MTA required
PARP1 Chromobody TagGFP TagRFP Sold in plasmid format; MTA required
Vimentin Chromobody TagGFP Sold in plasmid format; MTA required

Our Nanobody based Live cell imaging reagent are reagents are 1/10 the size of a conventional primary antibodies or IgG. The small size considerably improves tissue penetration and minimizes the distance between epitope and label, which increases imaging resolution and can make signals less diffuse. Hence, these reagents are perfectly suited for any imaging application, especially super-resolution microscopy techniques. primary nanobody epitope label displacement

With the monovalent nature of our Nanobodies deliver results as close to the native state as possible. Especially for cell surface receptors, this feature helps to avoid experimental bias due to dimerization and activation of the target protein.

ChromoTek’s recombinant Nano-Secondary reagents are characterized by both DNA and protein sequence. Therefore, we generate identical Nano-Secondary reagents in each manufacturing batch with great purity, and virtually no batch-to-batch variation. The production process of our nanobodies is completely animal free. Through site specific conjugation, we can produce nanobodies with a defined degree of labeling. We validate each batch i.a. with UV/VIS spectroscopy, SDS-PAGE, IF, and mass-spectrometry.