IF-Positive Antibody Discovery Service

Overview Principle Featured Services Key Features Published Data Precautions

Immunofluorescence (IF) is commonly used to evaluate the localization of proteins of interest as well as endogenous expression levels. It is worth noting that IF also plays an important role in the screening of hybridoma cells in antibody manufacturing. Leveraging extensive experience in antibody development, Creative Biolabs provides IF hybridoma screening services through our innovative hybridoma preparation and validation platform.

Introduction to IF

Fluorescently labeled antibodies are a critical factor in successful IF experiments. Similar to immunohistochemistry (IHC) and immunocytochemistry (ICC), IF is suitable for detecting and visualizing target proteins in fixed tissues or cells. It is worth noting that IF detection relies on fluorescent labels and lasers to generate signals.

How Does IF Work?

The principle of IF is based on the antigen-antibody response. Labeling antibodies or antigens with reactive fluorescent pigments is a crucial step. After the labeled antigen or antibody binds to the corresponding target, the characteristics and position of the target are determined under a fluorescent microscope. IF relies on specific primary antibodies and fluorescent dye-conjugated secondary antibodies to label cellular proteins. In the direct method, the conjugated primary antibody can also be used directly for labeling. At Creative Biolabs, our antibodies suitable for IF detection undergo rigorous testing. At Creative Biolabs, we are committed to screening hybridoma clones for IF applications, which is a critical step in the eventual generation of antibodies suitable for IF.

Fig.1 IF and H&E. (Lin, et al., 2023) Fig.1 Same-section IF and H&E staining.¹

Featured Services

Antibody performance is tested in appropriate tissues.
Cells with known target expression levels are used to ensure the specificity of the antibody.
The signal-to-noise ratio is measured to ensure excellent sensitivity.
Modified fixation and permeabilization conditions ensure antibody quality.
Sophisticated testing ensures batch-to-batch reproducibility.

Key Features

Highly specificity
High sensitivity
Rapid and concise process
Wide range of applications

Published Data

Images of IHC and uniplex IF. Fig.2 Microphotographs of representative examples of validation from uniplex IHC and uniplex IF.²

After conducting chromogenic IHC and uniplex IF staining to examine all the markers, Parra, Edwin R., et al. further validated these markers through multiplex IF staining in two distinct panels². To substantiate their findings, they presented microphotographs showcasing representative instances of validation derived from uniplex IHC, uniplex IF staining that highlighted tumor-associated immune cell expression, and intricate details of uniplex IF staining in tonsil tissue.

Precautions

The development of IF antibodies is challenging because the reagents used require high specificity for the target.
When different antibody conjugates are used in the same assay, it is particularly difficult to validate different antibody combinations to ensure easy interpretation of the data.
The quality of the antibody also plays a decisive role in IF imaging.

Our extensive experience in antibody development and optimized antibody functional verification strategies allow us to provide you with high-quality IF antibody products. Our functional validation ensures the specificity and affinity of the desired antibody. If you are in urgent need of cost-effective IF antibody development services, please feel free to contact us.

References

Lin, Jia-Ren, et al. "High-plex immunofluorescence imaging and traditional histology of the same tissue section for discovering image-based biomarkers." Nature Cancer 4.7 (2023): 1036-1052. Distributed under Open Access license CC BY 4.0. The image was modified by extracting and using only a and b sections of the original image.
Parra, Edwin R., et al. "Validation of multiplex immunofluorescence panels using multispectral microscopy for immune-profiling of formalin-fixed and paraffin-embedded human tumor tissues." Scientific reports 7.1 (2017): 13380. Distributed under Open Access license CC BY 4.0, without modification.

All listed services and products are For Research Use Only. Do Not use in any diagnostic or therapeutic applications.

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