Custom Services

Fluorophore Conjugation

Fluorophore conjugation refers to the process of attaching a fluorophore, a molecule that emits fluorescence upon excitation, to another molecule, typically a biomolecule such as proteins, antibodies, nucleic acids, or small molecules. This conjugation allows for the labeling and visualization of the target molecule using fluorescence-based techniques.

Design of Fluorophore Conjugation

• Choose a fluorophore based on your experimental needs. Consider factors such as emission and excitation spectra, photostability, and compatibility with your detection system.
• Employ suitable conjugation chemistry to label fluorophores to slected proteins or antibodies.

Applications of Fluorophore Conjugation

1. Immunofluorescence (IF) and Imaging

   Visualize the spatial distribution of specific proteins or antigens within cells or tissues using fluorophore-conjugated antibodies.

2. Flow Cytometry

   Quantify and analyze cell populations based on surface or intracellular marker expression using fluorophore-conjugated antibodies.

3. Western Blotting

   Detect and quantify specific proteins in a sample by using fluorophore-conjugated secondary antibodies for chemiluminescent or fluorescent detection.

4. Immunohistochemistry (IHC)

   Study protein localization and expression in tissue sections using fluorophore-conjugated antibodies.

5. ELISA and Multiplex Assays

   Implement fluorophore-conjugated antibodies in enzyme-linked immunosorbent assays (ELISA) or multiplex assays for high-throughput detection of multiple targets.

6. Live Cell Imaging

   Track dynamic cellular processes by using fluorophore-conjugated antibodies in live cell imaging experiments.

Custom Services of Fluorophore Conjugation

At InnoCyto Inc, we offer high quality, efficient and cost-saving antibody/protein fluorescent conjugation services to meet your specific requirements and experimental needs. We also provide functional test services from your needs.

Here are highlighted services:

• Labelling various fluorophores to antibodies or proteins.
• Dual labelling of fluorophores and oligonucleotides to antibodies or proteins.
• Small chemical fluorescents labeling, including FITC, Alexa Fluor, Cy dyes and many other dyes
• Fluorescent protein labeling, including green fluorescent protein (GFP, EGFP), red fluorescent protein (RFP), R-phycoerythrin (R-PE), B-phycoerythrin (B-PE), allophycocyanin (APC) and peridinin- chlorophyll-protein (PerCP) and derived tandems
• Labelling fluorophores to your specific biomolecules.

Oligonucleotide Conjugation

Oligonucleotide-Antibody conjugates have been used in numerous applications such as signal cell analysis and special genomic study from research diagnostics to therapeutic. we offer custom oligonucleotide conjugation services. The technology allows the conjugation of oligonucleotides (DNA and RNA) to the molecule of your choice.

Oligonucleotide conjugation refers to the covalent attachment of oligonucleotides (short sequences of nucleotides, typically DNA or RNA) to other molecules, such as proteins, peptides, or nanoparticles. This conjugation is achieved through various chemical methods, allowing the integration of oligonucleotides into diverse applications.

Design of Oligonucleotide Conjugation

• Choosing the appropriate oligonucleotide that is complementary to the target of interest.
• Employing specific conjugation methods to label oligonucleotide to your selected protein, antibodies or microbeads.
• For dual labelling, conjugating additional fluorophores or report molecules for visualization or detection purposes.

Applications of Oligonucleotide Conjugation

1. Enhanced Target Detection

   Oligonucleotide-conjugated antibodies or proteins can be used for enhanced signal detection in immunoassays. The oligonucleotides may carry multiple reporter molecules, amplifying the signal and improving sensitivity.

2. DNA/RNA Detection

   Oligonucleotide-conjugated proteins or antibodies can be employed for the detection of specific DNA or RNA sequences. This is valuable in molecular diagnostics, genotyping, and detecting infectious agents.

3. Cellular Imaging and Tracking

   Oligonucleotide-conjugated antibodies can be used to label and track specific proteins or cells in live cell imaging experiments. This is crucial for studying dynamic cellular processes.

4. Enzyme-Linked Oligonucleotide Assays (ELONA)

   Conjugating oligonucleotides to proteins can be used in ELONA for sensitive detection of specific analytes. This approach combines the specificity of antibodies with the amplification potential of oligonucleotides.

5. Multiplexing and Assay Development

   A set of unique oligonucleotide can be conjugated with distinct biomolecules, enabling multiplexed assays. This is valuable for simultaneously detecting multiple targets in a single sample.

6. Single Cell Sequencing

   Oligonucleotide conjugation is a crucial step in single-cell sequencing workflows, especially in methods like single-cell RNA sequencing (scRNA-seq). The conjugation process involves attaching specific oligonucleotide sequences to individual cells, enabling the identification and profiling of gene expression at the single-cell level.

Custom Services of Oligonucleotide Conjugation

At InnoCyto Inc, we offer oligonucleotide-antibody/protein conjugation by our expertise in the field with a successful track record. All antibody/protein oligonucleotide conjugates are analyzed in order to remove unconjugated antibody/protein. We also offer antibody/protein-fluorescent-oligonucleotide dual labelling conjugation to allow you to study both protein expression and genomics simultaneously.

Here are highlighted services:

• Labelling oligonucleotides to antibodies or proteins.
• Dual labelling of fluorophores and oligonucleotides to antibodies or proteins.
• Labelling oligonucleotides to microbeads for selection and isolation of target DNA sequences.
• Labelling oligonucleotides to your specific molecules.

Streptavidin & Biotin Conjugation

Streptavidin is a tetrameric protein that has a high affinity for biotin, a small molecule. The interaction between streptavidin and biotin is one of the strongest non-covalent interactions in nature.

Biotin can be conjugated to various molecules, including proteins, antibodies, and nucleic acids. This conjugation is often achieved through chemical methods, resulting in a covalent bond between the biotin molecule and the target molecule.

Design of Streptavidin and Biotin Conjugation

For biomarker detection, Labelling Biotin to proteins or antibodies. For solid phase or isolation, attaching Streptavidin to microbeads.

Applications of Streptavidin and Biotin Conjugation

1. Immunohistochemistry (IHC) and Immunofluorescence (IF)

   Biotinylated antibodies are often used in immunohistochemistry and immunofluorescence assays. Streptavidin is then used as a detection reagent by binding to the biotinylated antibodies, facilitating the visualization of specific proteins.

2. Western Blotting

   Biotinylated probes (e.g., antibodies or streptavidin itself) can be used in Western blotting to detect specific proteins. Streptavidin-horseradish peroxidase (streptavidin-HRP) or streptavidin-alkaline phosphatase (streptavidin-AP) is commonly employed for signal amplification.

3. Fluorescence In Situ Hybridization (FISH)

   Biotinylated nucleic acid probes can be used in FISH experiments. Streptavidin conjugated to fluorophores is then applied to detect the biotin-labeled probes, allowing visualization of specific DNA or RNA sequences.

4. Enzyme-Linked Immunosorbent Assay (ELISA)

   Biotinylated antibodies are frequently used in ELISA. Streptavidin-HRP or streptavidin-alkaline phosphatase is added to bind to the biotinylated antibodies, resulting in signal amplification for sensitive detection.

5. Cell Sorting and Flow Cytometry

   Biotinylated antibodies can be used for labeling specific cell surface markers. Streptavidin-conjugated fluorophores are then applied for detection and sorting of labeled cells in flow cytometry.

6. Purification of Biotinylated Molecules

   Streptavidin is commonly used in affinity purification to capture and isolate biotinylated proteins, peptides, or nucleic acids.

Custom Services of Streptavidin and Biotin Conjugation

The streptavidin-biotin interaction is versatile and widely utilized in various biological and biomedical applications due to its high specificity and stability. Researchers and companies often use streptavidin-biotin conjugation for designing custom assays and detection systems.

At InnoCyto Inc, we offer specialized custom services for streptavidin and biotin conjugation tailored to meet your unique research and experimental needs. Our team of experts is dedicated to providing high-quality conjugation solutions, ensuring precise and efficient binding of streptavidin or biotin to target proteins, antibodies or microbeads.

Here are highlighted services:

• Labelling biotin to proteins or antibodies.
• Labelling streptavdin to microbeads.
• Labelling streptavdin or biotin to your specific molecules.

Horseradish Peroxidase (HRP) Conjugation

HRP conjugation involves linking horseradish peroxidase, an enzyme, to another molecule, often a biomolecule or a probe. This conjugation is commonly employed in various assays and techniques for detection and visualization purposes.

Design of HRP Conjugation

• Choose the target molecule based on the application (e.g., antibodies for immunoassays, proteins, or nucleic acids).
• Activate HRP for conjugation by introducing suitable functional groups on its surface.
• Purify the HRP conjugate to remove any unconjugated molecules and ensure the desired specificity.

Applications of HRP Conjugation

Immunoassays

LHRP-conjugated antibodies are widely used in immunoassays like ELISA (Enzyme-Linked Immunosorbent Assay) for detecting the presence of specific antigens. Facilitates colorimetric, chemiluminescent, or fluorometric detection methods.

Immunohistochemistry (IHC) and Immunocytochemistry (ICC)

LUsed for visualizing the localization of specific proteins in tissues or cells. Enables the development of colored or fluorescent signals in microscopy.

Western Blotting

LHRP-conjugated secondary antibodies are employed to detect primary antibodies bound to target proteins. Facilitates chemiluminescent or colorimetric detection in Western blot analyses.

In Situ Hybridization (ISH)

LHRP-conjugated probes can be used to detect specific nucleic acid sequences in cells or tissues. Allows for the visualization of gene expression patterns.

Enzyme-Linked ImmunoSpot (ELISpot) Assays

LUsed for quantifying the number of cytokine-secreting cells. HRP-conjugated antibodies enable the visualization of spots corresponding to cytokine-producing cells.

Custom Services of Particle Bead Conjugation

InnoCyto offer specialized custom services for Horseradish Peroxidase (HRP) conjugation tailored to meet your unique research and experimental needs. Our team of experts is dedicated to providing high-quality conjugation solutions, ensuring precise and efficient binding of HRP to biomolecules. or nanoparticles.

Here are highlighted services:

• Labelling HRP to antibodies.

Particle Bead Conjugation

Particle bead conjugation involves attaching biomolecules such as antibodies, proteins or small molecules) to microbeads. The conjugation is typically achieved through covalent or non-covalent binding, allowing the beads to carry specific molecules for various applications.

Design of Particle Bead Conjugation

• Choose magnetic or polystyrene microbeads with a suitable size for your application.
• Attach your target proteins or small molecules to microbeads.
• Wash the beads throughly to remove any unbound target proteins or small molecules.

Applications of Particle Bead Conjugation

1. Immunoprecipitation (IP) and Co-Immunoprecipitation (Co-IP):

   Antibodies can be conjugated to beads to selectively pull down specific proteins from a complex mixture. This is used in immunoprecipitation and co-immunoprecipitation experiments for protein-protein interaction studies.

2. Cell Sorting and Flow Cytometry:

   Beads can be conjugated with antibodies or other ligands to label specific cell surface markers. This enables the isolation and sorting of specific cell populations in flow cytometry.

3. Magnetic Bead Separation:

   Magnetic beads can be conjugated with antibodies or other ligands to selectively capture and separate target molecules or cells using a magnetic field. This is commonly used in cell isolation and purification.

4. Solid Phase Support

   Solid Phase Support: Beads can serve as solid phase supports in Enzyme-Linked Immunosorbent Assay (ELISA). They can be conjugated with capture antibodies or antigens, providing a platform for sensitive and specific detection of target molecules.

5. Nucleic Acid Purification:

   DNA/RNA Isolation: Beads can be coated with substances that specifically bind nucleic acids. This is used in various nucleic acid purification methods, such as DNA/RNA extraction and isolation.

6. Multiplexing and Assay Development

   Different types of beads can be conjugated with distinct biomolecules, enabling multiplexed assays. This is valuable for simultaneously detecting multiple targets in a single sample.

   Conjugated beads are employed in various diagnostic assays for the detection of biomarkers, pathogens, or disease-related molecules in clinical samples. Bead conjugation provides a versatile platform for various bioanalytical and biomedical applications, offering flexibility in designing custom assays and detection systems.

Custom Services of Particle Bead Conjugation

InnoCyto offer specialized custom services for particle bead conjugation tailored to meet your unique research and experimental needs. Our team of experts is dedicated to providing high-quality conjugation solutions, ensuring precise and efficient binding of biomolecules to microbeads or nanoparticles.

Here are highlighted services:

• Labelling specific antibodies to microbeads.
• Labelling Streptavidin to microbeads.
• labelling oligonucleotides to microbeads.

Dual Labelling - Fluorophore plus Oligonucleotide

The process conjugates both a fluorophore and an oligonucleotide to an antibody or protein, is commonly known as "dual labeling" or double labeling." This dual labelling technique allows researchers to simultaneously detect the presence of the antibody and study the localization or expression of a specific target molecule.

Design of Dual Labelling Conjugation

• Antibody Conjugation with Fluorophore - attaching a fluorophore to the antibody.
• Oligonucleotide conjugated to Fluorophore-labelled Antibody - labelling an oligonucleotide to the fluorophore. Or alternatively labelling an oligonucleotide to the antibody per your request.
• Purification and Validation - purifying and validating the dual labelling antibody products.

Applications of Dual Labelling Antibodies

Dual labeling involving both an oligonucleotide (oligo) and a fluorophore on an antibody provides a versatile tool for various applications. The dual-labeled antibody is introduced to the cells, allowing it to bind both to the target protein and to the mRNA transcripts in the cells. Here are some common applications of dual-labeled oligo and fluorophore antibodies:

1. Fluorescence In Situ Hybridization (FISH) with Antibodies (Immuno-FISH)

   This technique combines the specificity of antibodies with the ability to visualize specific nucleic acid sequences using FISH. Dual-labeled antibodies enable the detection of both proteins and nucleic acids in the same sample, providing information about their colocalization.

2. Cellular Imaging and Localization

   Dual-labeled antibodies can be used to study the subcellular localization of specific proteins along with the visualization of corresponding mRNA. This provides insights into cellular processes and organization.

3. Single-Cell Analysis

   For single-cell studies, dual labeling allows the detection of both proteins and specific RNA sequences within individual cells. This is useful for understanding heterogeneity in gene expression at the single-cell level.

4. Multiplexed Assays

   Dual labeling facilitates multiplexed assays where researchers can detect multiple proteins and nucleic acid targets simultaneously. Different fluorophores and oligonucleotides enable the detection of a diverse set of targets in a single experiment.

5. Molecular Profiling of Tissues

   In studies involving tissues, dual labeling helps profile molecular markers by simultaneously visualizing protein expression and the presence of specific RNA sequences. This is valuable in cancer research and other disease studies.

6. In Situ PCR and Antibody Staining

   Dual-labeled antibodies can be used in combination with in situ PCR to detect specific DNA sequences along with the corresponding protein expression in situ. This approach allows for detailed molecular characterization of cells.

Custom Services of Dual Labelling Conjugation

With strong experise and advanced technology, we offer custom service of dual labeling conjugation to satisfy your specific requirements and applications. Here are some common dual labelling solutions in our custom services:

• Dual labelling conjugation of fluorophore and oligonucleotide to Antibody
• Dual labelling conjugation of fluorophore and oligonucleotide to Protein
• Fluorescence and oligonucleotide conjugation

This dual labelling solution allows you to simultaneously study both protein expression and genomics. Whether you require simultaneous detection of proteins and nucleic acids, multiplexed assays, or specialized imaging studies, our expertise in dual labeling enables us to deliver customized solutions for a wide range of applications. Explore the possibilities of our comprehensive conjugation services designed to enhance the specificity, sensitivity, and versatility of your experiments.