Immunofluorescence
Empowering discovery with functionalized tools for fluorescence-based detection
What is Immunofluorescence (IF)?
Immunofluorescence (IF) is a powerful and versatile technique in biological research that enables scientists to visualize proteins and other antigens within cells and tissues. By combining the specificity of antibodies with the properties of fluorescent dyes, IF allows for the precise localization of target antigens making it an essential tool for studying biomarkers, protein expression, cellular architecture, disease mechanisms, and other biological processes.
Immunofluorescence is an indispensable technique for multiplexing, enabling the simultaneous detection of multiple targets within a single sample while preserving spatial context. It is widely used in cancer research, neuroscience, immunology, and infectious disease studies to examine cellular interactions and tissue organization, identify pathological changes and track disease progression, and analyze signaling pathways. Unlike immunohistochemistry (IHC), IF allows for the detection of multiple antigens within the same cell and even within specific cellular compartments, providing more precise insights into colocalization and molecular interactions. By offering high-resolution spatial and molecular data, IF supports a deeper understanding of complex biological systems in health and disease.
Vector Laboratories’
Immunofluorescence Reagents
Vector Laboratories provides comprehensive, end-to-end solutions to inable every step of the immunofluorescence process. Our portfolio supports each stage of the IF workflow, from sample preparation to imaging, ensuring high-quality, reproducible results in drug discovery, translational medicine, and biomarker validation.
At Vector Laboratories, we are committed to delivering high-quality reagents and unparalleled support to ensure the success of your IF workflows. Here are the key features that set us apart:
Scalable, Precision-Driven Manufacturing
Our optimized and scalable manufacturing processes deliver consistent, high-performing reagents at any scale, providing a reliable foundation for reproducible and impactful results.
Robust Quality
Control (QC)
Rigorous intended-use QC testing ensures batch-to-batch consistency, giving researchers confidence in every experiment and enabling accurate, reliable outcomes.
Proven
Stability
Our reagents offer exceptional stability, ensuring reliable performance throughout their shelf life and minimizing experimental variability.
Quality
Management System
Rigorous intended-use QC testing ensures batch-to-batch consistency, giving researchers confidence in every experiment and enabling accurate, reliable outcomes.
Benchmarking Performance
Vector Laboratories’ reagents deliver industry-leading performance, setting the gold standard in IF workflows and providing researchers with superior quality data.
Dedicated
Technical Support
Our expert support team is available to assist with assay optimization, troubleshooting, and technical guidance, ensuring your success at every step of the workflow.
Trusted and Cited
in Publications
Our products are widely trusted and cited in high-impact scientific journals, highlighting their credibility and reliability in advancing groundbreaking research.
Immunofluorescence Workflow
Slide Preparation
Sections should be mounted on charged slides to maximize tissue section retention and adherence on glass slides. Demarcate and isolate individual sections to reduce reagent use or for discrete treatment.
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Antigen Unmasking
Heat-induced epitope retrieval (HIER) using buffers like citrate, Tris, or EDTA is often required to unmask antigens that may be cross-linked during fixation, enhancing antibody accessibility.
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Blocking
Reduce or eliminate unwanted background (non-specific) staining on tissue sections and cell preparations using a blocking solution. Non-specific staining can result from endogenous enzyme or tissue elements, including endogenous enzyme activity, presence of Fc receptors, or interactions of detection reagents with tissue or cell proteins and other macromolecules. Choose a blocking solution based on the results of negative control sections.
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Primary Antibody / Lectin
Identify and localize the target protein antigen or glycoprotein moiety in cells or tissue preparations using specific validated markers. When choosing a primary antibody or lectin, consider the tissue species and tissue preparation method (e.g., fixation) to ensure specific binding of your target.
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Secondary Antibody
These affinity-purified, optimally conjugated, and highly specific antibodies are directed against the most commonly used primary antibody target species and are available with a choice of fluorophores
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Tertiary Reagent
If needed, increase the sensitivity of detection to visualize weakly or transiently expressed, upregulated, or unknown (e.g., gene knock-in studies) antigens. Remove antibodies and other non-covalently bound molecules for additional detection.
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Autofluorescence Quencher
Remove unwanted fluorescence in tissue sections due to aldehyde fixation, red blood cells, and structural elements such as collagen and elastin.
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Counterstain / Mount
Retain and preserve fluorescent signal intensity, with or without optional counterstain, to increase microscopy exposure times and archive for future imaging and reference requirements.
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Featured Product Categories
We develop products within the following product categories to enable researchers to provide comprehensive, end-to-end solutions to optimize every step of the immunofluorescence (IF) process. With reagents that streamline antigen retrieval, block unwanted fluorescence, enhance target detection, and provide antifade mounting of your precious tissues, our catalog supports the IF workflow from start to finish. Additionally, we provide functionalized solutions for challenges like species-on-species detection and multiplexing so researchers can achieve exceptional sensitivity, specificity, and reproducibility in even the most advanced IF applications.
Primary Antibodies
Successful immunofluorescence outcomes are highly dependent on the specificity and sensitivity of the primary antibody, and not all antibodies are equal to the task. The best antibodies for IF must be able to perform in IHC applications and demonstrate a strong signal with minimal background noise in a variety of tissue types.
Vector Laboratories provides a prolific product portfolio of validated and highly cited primary antibodies for use in immunofluorescence, including IHC-Plus™ and PathPlus™ antibodies from our LSBio brand that are validated in 20+ tissue types for optimal performance, high affinity goat polyclonal antibodies from our Everest Biotech brand,and recombinant engineered antibodies from our Absolute Antibody brand.
Recombinant Antibodies
Recombinant antibodies are manufactured in vitro by Absolute Antibody using synthetic genes with 100% biological definition to ensure the highest reproducibility and purity. Recombinant technology enables the engineering of antibodies into new species, isotypes, and formats to suit application requirements. Recombinant monoclonal antibodies can serve as highly specific primaries for immunofluorescence and can be customized for effective multiplexing.
Recombinant antibody technology ensures:
- Reproducibility
- Biological definition
- High purity reagents
- Multiplexing capabilities
Solutions for Advanced IF Workflows
Immunofluorescence workflows can be complex and technically demanding requiring specialized approaches to reduce autofluorescence background, eliminate interference from endogenous immunoglobulins, and enable high-resolution multiplexing. Vector Laboratories offers industry-leading solutions to overcome common IF challenges such as autofluorescence, low signal-to-noise, and complex multiplexing workflows to empower researchers to successfully execute their advanced IF experiments and achieve optimal results.
Mouse-on-Mouse Detection
Vector’s Mouse on Mouse (M.O.M.®) Kits are specifically designed for detecting and visualizing unconjugated mouse primary antibodies in mouse tissue specimens. These kits contain a proprietary reagent that blocks endogenous mouse Ig in specimens therefore preventing non-specific binding of subsequent anti-mouse IgG detection reagents. Explore our Fluorescein Kit (FMK-2201) and other species-on-species technologies.
Autofluorescence
Tissue autofluorescence often occurs with aldehyde fixation or from inherent native tissue components (collagen, elastin, and red blood cells). The extent and intensity of autofluorescence background frequently makes it difficult or impossible to distinguish specific signals in immunofluorescence applications. The TrueVIEW Autofluorescence Quenching Kit provides a novel way to diminish unwanted autofluorescence from non-lipofuscin sources and dramatically improve signal-to-noise ratio.
Multiplexing Solutions
Multiplex immunofluorescence (mIF) enables researchers to detect multiple biomarkers within a single tissue sample providing deeper insights into cellular signaling, immune responses, and tumor microenvironments. This technique is especially valuable in oncology, immunotherapy research, and spatial transcriptomics where understanding protein colocalization and tissue architecture is critical or when sample availability is limited. However, mIF presents challenges such as cross-reactivity, reagent compatibility, and potential tissue damage from repeated staining cycles. The VectaPlex™ Antibody Removal Kit simplifies multiplex workflows by efficiently removing antibodies and other non-covalently bound detection reagents facilitating assay design and optimization while preserving tissue antigenicity and sample integrity.
Immunofluorescence Research Areas
Immunofluorescence is a powerful tool that allows researchers to visualize, characterize, and quantify structures and pathways to enhance our understanding of areas like cancer, neuroscience, drug development, and more. By providing a functionalized portfolio of IF reagents and antibodies, we power research across diverse disciplines.
Cell Biology
Immunofluorescence visualizes protein distribution, cytoskeletal organization, and organelle dynamics, allowing for study of cellular processes.
Drug Discovery & Development
IF aids in the visualization and quantification of target proteins, cellular responses, and biomarker expression, helping to assess drug efficacy and mechanism of action.
Immunology
Mapping immune cell interactions, detecting specific antigens, and tracking signaling pathways with IF reveals disease mechanisms and immune function.
Infectious Disease Research
IF supports infectious disease research by detecting pathogens, visualizing host-pathogen interactions, and monitoring immune responses at the cellular level.
Developmental Biology
IF enables researchers to track protein expression, visualize cell differentiation, and analyze tissue organization over time.
Neuroscience Research
Immunofluorescence is used to label and visualize neural structures, track protein expression, and study cellular interactions in the nervous system.
Oncology Research
From identifying cancer biomarkers, analyzing tumor microenvironments, and assessing treatment responses at the cellular level, IF supports oncology research.
Immunofluorescence Research Areas
Immunofluorescence is a powerful tool that allows researchers to visualize, characterize, and quantify structures and pathways to enhance our understanding of areas like cancer, neuroscience, drug development, and more. By providing a functionalized portfolio of IF reagents and antibodies, we power research across diverse disciplines..
Resources
Immunofluorescence Resource Guide
Streamline the immunofluorescence process with this step-by-step guide to finding the optimal reagents for your project
Immunofluorescence Mounting Guide
Explore the broad spectral coverage, unique performance characteristics, and reliable performance of fluorophores from Vector Laboratories.
