IHC Staining: Targeted Protein and Biomarker Detection in Tissue Sections

Overview

IHC staining (Immunohistochemistry) is a powerful technique for detecting specific proteins, antigens, or biomarkers within formalin-fixed, paraffin-embedded (FFPE) tissue sections. By using antigen-specific antibodies and chromogenic or fluorescent detection, IHC enables precise spatial localization of cellular targets.

At iHisto, our IHC staining services combine validated protocols, automated platforms, and expert antibody optimization to deliver high-sensitivity, high-specificity results for academic, biotech, and preclinical clients.

Whether you're identifying disease markers, validating targets, or characterizing therapeutic effects, our IHC workflow ensures clarity and reproducibility.

How IHC Staining Works

IHC uses primary antibodies to bind specific protein targets in tissue. Detection is achieved using enzyme-conjugated secondary antibodies (e.g., HRP or AP) and a chromogenic substrate such as DAB, which creates a colored precipitate at the antigen site.

Color Results (Typical DAB protocol):

• Target protein: Brown (DAB signal)

• Nuclei: Blue (with hematoxylin counterstain)

• Cytoplasm / background: Clear or lightly stained

Other chromogens or fluorescent labels can be used depending on multiplexing or sensitivity needs.

Step-by-Step IHC Staining Process

1. Antigen Retrieval
   Heat- or enzyme-induced retrieval is performed to expose epitopes.

2. Blocking
   Reduces non-specific binding of antibodies and endogenous enzyme activity.

3. Primary Antibody Incubation
   Tissue is incubated with a validated primary antibody specific to the target protein.

4. Secondary Detection
   HRP- or AP-linked secondary antibodies bind the primary antibody.

5. Chromogenic Reaction
   Substrates like DAB or AEC are applied to visualize the antigen-antibody complex.

6. Counterstaining and Mounting
   Hematoxylin is added for nuclear contrast; slides are then dehydrated and coverslipped.

Our Optimized IHC Workflow

iHisto’s IHC staining services are supported by:

• Leica BOND RX or Thermo Autostainers – Automated, high-throughput platforms

• Antibody optimization – Titration, retrieval, and incubation condition testing

• Validated controls – Positive and negative controls included for each batch

• Digital slide scanning – High-resolution output for review, annotation, or AI analysis

We support both single-plex and multiplex chromogenic or immunofluorescent staining formats.

Applications of IHC Staining

IHC is essential in translational research and clinical development for spatially mapping protein expression.

Common use cases include:

• Oncology – Tumor subtype characterization, proliferation markers (e.g., Ki-67), immune markers (CD3, CD8, PD-L1)

• Neuroscience – Glial, neuronal, and synaptic markers (e.g., GFAP, NeuN)

• Toxicology – Tissue-specific damage and biomarker response

• Inflammation – Cytokines, macrophages, lymphocytes

• Biomarker validation – For preclinical therapeutic studies

Whether you're validating an antibody or profiling tumor response, IHC staining provides reliable, localized expression data.

Advantages of iHisto’s IHC Staining Services

• ✅ Wide panel of validated antibodies and detection systems

• ✅ Chromogenic and fluorescent IHC available

• ✅ Human, rodent, and non-human primate tissue supported

• ✅ High-quality counterstaining and signal-to-noise control

• ✅ Optional pathology review, quantification, and slide digitization

We offer flexible IHC services for discovery, GLP-compliant preclinical, and biomarker development programs.

FAQs

• What tissues can you perform IHC on?
  We work with FFPE sections from human, mouse, rat, NHP, and other models — including tumor, brain, liver, and immune organs.

• Do you help with antibody optimization?
  Yes. We test and validate new antibodies with multiple titrations and retrieval conditions for optimal staining.

• Can I request multiple markers on the same slide?
  Absolutely. We offer multiplex chromogenic and fluorescent IHC, including custom marker panels.

• Do you digitize slides for image analysis?
  Yes. We provide whole-slide imaging, along with optional AI-assisted quantification and cloud delivery.

Ready to Partner with an Expert IHC Lab?

At iHisto, we specialize in high-performance IHC staining for academic research, biotech, and drug development. Whether you’re screening antibodies or validating disease targets — we deliver clear, reproducible results you can trust.

📩 Contact us to request a quote or schedule a consultation.