Immunohistochemistry & Immunofluorescence – Biomarker Analysis on Protein level

Immunohistochemistry (IHC)

Our IHC standard services

Immunohistochemistry (IHC) is commonly used to analyse biomarkers in different type of tissue sections on protein level.

At TPL Path labs a team of highly experienced scientists and technicians is carefully considering every step of your IHC project. Integrating all available information of your samples and the antibodies of choice, understanding your particular demands, but also taking into account potential challenges to find the best approach to visualize your target of interest – right from the start!

Our expertise covers state of the art immunohistochemistry on in-vitro cell cultures, diverse animal models and human tissue samples. As a result of a multitude of successfully accomplished projects, as well as projects currently going on we rely on a constantly growing portfolio of validated protocols for a variety of targets to label tissues.

We are ready to get started

Utilising state-of-the-art Ventana Discovery Ultra and BenchMark Ultra platforms, we are ready to visualise the following well established diagnostic targets in your samples:

Oncology/proliferation/apoptosis

  • Caspase 3 – apoptosis
  • Ki67 – proliferation marker
  • TUNEL assay – apoptosis
  • TNFRSF10B/Killer/DR5 – apoptosis

Organelles and tissue types

  • Lamp 2 – Lysosomal and late endosome marker
  • CD31 – endothelial marker
  • Chromogranin A – tumor marker

Immunology/Immuno-phenotyping

  • CD3e – T-cells
  • CD4 – T-helper cells
  • CD8 – cytotoxic T-cells
  • CD14 – monocytes marker
  • CD20 – B-lymphocytes
  • CD21 – B-cells
  • CD23 – B-cells
  • CD45 – Leucocytes
  • CD68 – Macrophages
  • CD79A – B-cells

In addition to the ones listed above, we have experience with a whole range of antibodies against the targets listed below. Please do not hesitate to inquire for your specific project!

Immunology and Inflammation

  • 5 Lipoxygenase (5LO) – inflammatory processes
  • BCL6 – germinal center T and B cells/ primary immature dendritic cells
  • BD2 (Beta-defensin 4A) – Inflammation – antimicrobial activity
  • CDC27 – Plasmacells, Effector B-cells, T-cells
  • CD1A – Thymocytes, dentritic cells
  • IL-17 – Cytokine
  • IL-6 – Cytokine
  • IL-1b – Cytokine
  • IL-18 – Cytokine
  • CD99 – T-cells
  • CD74 – Macrophages/B-cells
  • CD10 – Immature B-cells, lymphoid precursor cells, Granulocytes
  • CD1A – Thymocytes, dentritic cells
  • CD68 – Macrophages
  • CD43 – Thymocytes; T-Lymphocytes
  • CD5 – T-/ B-cell subpopulations
  • CD56 – NK cells
  • MPO – Immunology/Lysosome
  • Kappa light chain – Immunoglobulin
  • Lambda light chain – Immunoglobulin

Organelles and tissue types

  • Actin (Clone HHF35) – muscle cells/Cytoskeleton
  • EpCAM (clone BER-EP4) – Epithelial cell adhesion molecule
  • CD144 (VE-Cadherin) – Intercellular Junction Marker
  • CD138 – Syndecan-1, Cell surface proteoglycan, links the cytoskeleton to the interstitial matrix
  • DES Desmin – Cytoskeleton Marker
  • Myosin9 – Cytoskelleton
  • Podoplanin (PDPN) – Cytoskelleton/ Cell migration
  • PLEC – Cytoskelleton/ Cell migration
  • STMN1 – Cytoskelleton
  • Tubulin (acetylated) – Cytoskelleton
  • Tubulin (β3) – Cytoskelleton
  • Tubulin Detyrosinates (Glu-Tu) – Cytoskelleton
  • VIM – Cytoskelleton/ Cell migration
  • α-SMA – Cytoskelleton/ Cell migration, blood vessels

Immunology/Immuno-phenotyping

  • ADK (Adenosine kinase) – Purine metabolism
  • BCL2 – Apoptosis regulator
  • BUB3 – Mitotic checkpoint protein
  • BUBR1- Mitotic checkpoint
  • CASP3 – Apoptosis
  • eIF4E Eukaryotic translation initiation factor 4E – Cell cycle/Transcription
  • p-eIF4E – Protein synthesis
  • EB1- modulator of APC activity
  • FTH1 Ferritin Heavy Chain – Iron metabolism
  • FTL Ferritin Light Chain – Iron metabolism
  • Iba1 – membrane ruffling and cell migration
  • MAP4 – Cytoskelleton (non-neuronal)
  • Mad2 – Cell cycle
  • VHL – von Hippel-Lindau

Neurology

  • CHRNA3 Neuronal acetylcholine receptor subunit alpha-3 – synapse, cell junction
  • CHRNA4 cholinergic receptor nicotinic alpha 4 subunit – synapse, cell junction
  • CHRNA7 Nicotinic Acetylcholine Receptor alpha 7 – synapse, cell junction
  • GFAP – Astrocyte marker
  • NSE – Neurology
  • S100 – Astrocyte marker

Cellular compartments and structures

  • CK18 Cytokeratin 18
  • CD34 – hematopoietic stem cells; endothelial cells
  • CD31 – endothelial marker
  • Lysozyme – Cell organelle
  • PMP70 – Peroxisome

Cancer/ Cancer related

  • Carcino Embryonic Antigen (CEA) – colon, pancreas, lung and breast
  • CHGA (Chromogranin A) – tumor marker
  • HER 2 (ErbB 2) – cancer
  • Melanoma [HMB45] – malignant melanoma marker
  • EMA (MUC1/tumor associated Mucin) – marker of breast cancer

Other

  • F13A Coagulation factor XIII A chain
  • GPX1 Glutathione Peroxidase 1
  • HHV8 (Human Herpes virus 8)
  • Melanin A – Pigment
  • MMP9 – Metalloprotease/extracellular matrix brakedown
  • Aquaporin 3 (AQP3) – epithelial cells of kidney (collecting ducts) and airways, in keratinocytes, immature dendritic cells and erythrocytes.
  • TTF1 – Transcription factor/Thyroid, lung

Our IHC custom tailored services

In addition to our standard IHC service we do offer custom tailored IHC services.

Our custom services include:

  • Antibody and target selection for your specific study
  • Companion diagnostic validation
  • Validation for translational and clinical research
  • Biomarker screening, using commercial and proprietary antibodies alike

Quality matters

Whether for our standard IHC services or for our custom tailored study – quality matters to us. To ensure the best possible validation of an antibody or a staining we do have the following controls available that can be included in your experiment if desired:

  • Isotype controls
  • Positive control tissues: We do have a large collection of various tissue controls derived from different species with complete organ sets that can be chosen and included as a positive control for your target of choice
  • Negative control tissue
  • Co-staining (double staining or consecutive sections) with an already established marker to evaluate the localisation of a so far not very well characterized target
  • Further validation of the expression of the target on mRNA level -> please click here for more information on our in situ-hybridization or DNA/RNA services

We never stop evolving …

We constantly keep an eye on novel developments in the field. Currently we are implementing a newly developed (Roche, Ventana Medical Systems) approach of multiplexing up to 5 antibodies, using chromogenic detection on our Ventana Discovery Ultra platforms.

Immunofluorescence (IF)

Fluorescence is a real alternative to chromogenic approaches to immunochemistry (IHC). Its High Dynamic Range makes it easier to visualise rare and highly abundant targets in protein localisation studies on the same slide. Moreover, it offers better visualisation of co-localisation of targets with multiple colour identification in multiplexing assays.

Due to the latest developments in fluorophore and microscope technology, the selection of colours used for both single- and multi-colour fluorescence microscopy are greater than ever.

We are highly experienced in direct or indirect detection methods on samples from cell culture, animal models and tissue. Our laboratory routinely uses fluorescent IHC detection to identify e.g. small amounts of biomarkers or to reveal precise subcellular localisation.