In vitro diagnostics (IVD) are chosen in recent years to detect and diagnose varieties of diseases, as well as determining the state of health, so as to mitigate, cure, and prevent diseases and their complications. Creative Biolabs is known as a famous expert in IVD development services and is good at manufacturing immunohistochemistry based kits to fit researchers’ project needs.

Immunohistochemistry

Immunohistochemistry (IHC) that utilizes monoclonal and polyclonal antibodies for the detection of specific antigens in tissue sections is an extremely powerful tool in the diagnostic and research laboratories. IHC requires the available biopsies which are processed into sections by a microtome and then incubated with an appropriate antibody. The antibody binding site is visualized under an ordinary or fluorescent microscope by a marker, for instance, fluorescent dye, radioactive element, enzyme, or colloidal gold, directly linked to the primary antibody or to a secondary antibody.

Immunohistochemistry staining. Fig.1 Immunohistochemistry staining. (Ward, J.M., 2014)

IHC plays an important role in the application of monoclonal and polyclonal antibodies to validate the tissue distribution of an antigen of interest in health and disease. It also plays a part in pathology, especially in the subspecialties of tumor pathology, neuropathology, and hematopathology. To date, IHC has widely used in the diagnosis of cancers since the expression of specific tumor antigens is up-regulated in some carcinomas. In autopsy subjects, the basic histologic examination by IHC is regarded as a useful and necessary component that may provide a greater insight.

The Workflow of IHC

The routinely experimental flow of IHC includes processes as follows:

  • Sample preparation
  • Immobilization
  • Tissue sectioning
  • Paraffin embedding
  • Blocking
  • Antigen retrieval
  • Detection
  • Counterstain
  • Mounting media

Currently, a new IHC assay named Opal mIHC is reported. Its protocol is very similar to the standard IHC, with the distinct exception being the iterative nature of the process enabled by a post-visualization microwave step in a citrate buffer. That offers a simplified multiplexing strategy, where the primary antibody is able to be chosen based on the performance, but not on species.

Opal multiplexed immunohistochemistry (mIHC) method.Fig.2 Opal multiplexed immunohistochemistry (mIHC) method. (Stack, E.C., 2014)

IHC-based Kits Development

IHC is a procedure that antibodies are used to detect proteins (antigens) of cells within tissue sections. The assay involves the staining of tissues, rather than cells, to determine the location of proteins of interest in a tissue sample. IHC staining requires very careful precision in slicing specimens, just allowing for one layer of cells on the slide. Afterwards, the resulting image by instruments demonstrates the concentration of proteins, showing the information regarding the biodistribution of compounds in a sample.

Creative Biolabs not only can offer machine-operated IHC staining services, but is skilled at manufacturing IHC-based kits for IVD study. We also provide optimization of experimental conditions for the primary antibody, like concentrations, incubation times, and interpretation of the staining results. There are cell line slides and tissues from different organs can be used for our customers if they do not have. The IHC kits of Creative Biolabs offer qualified reagents and validated tests to perform highly specific immunostaining for further biochemistry research. For more detailed information, please feel free to contact us.

References

  1. Ward, J.M., (2014). “Rodent immunohistochemistry: pitfalls and troubleshooting.” Vet Pathol, 51(1), 88-101.
  2. Stack, E.C., (2014). “Multiplexed immunohistochemistry, imaging, and quantitation: a review, with an assessment of Tyramide signal amplification, multispectral imaging and multiplex analysis.” Methods,70(1), 46-58


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