Immunohistochemistry (IHC) Experiment Service

Instruction manual

First Edition (Revised on April, 2016)

Available Services

1. Tissue embedding and slicing services.
2. Immunohistochemistry and immunofluorescence service for paraffin sections, frozen sections and adherent cells. Paraffin sections and frozen sections: Fixation or deparaffinization, antigen retrieval, blocking of endogenous peroxidase, incubation with primary antibody and secondary antibody, DAB coloration, counterstain with hematoxylin, sealing coverslip, observe and take photographs under microscope.
Adherent cells and immunofluorescence: Nuclear staining with DAPI, microscopic examination by inverted fluorescence microscope and image acquisition.

Samples for detection

1. Paraffin sections, frozen sections, and cells cultured in 6-well plates or cell culture dish.
2. Tissues. The embedding and slicing services can be provided.
3. In order to get good result in IHC test, please fix the embedded tissue or surgical specimens as soon as possible to prevent the tissues autolysis and antigen denaturation.

Service schedule

Service items	Sample types	Scheduled time
Common tissue embedding	Fixed tissue	2 workdays
Paraffin section	Paraffin embedded blocks	1 workday
Frozen section	Fixed tissue, fresh tissue	4 workdays
Immunohistochemistry	Slices, cells growing on glass coverslips	5 workdays
Immunofluorescence (single staining)	Slices, cells growing on glass coverslips	5 workdays
Immunofluorescence (double staining)	Slices, cells growing on glass coverslips	5 workdays

Materials provided by the customer

1. Paraffin sections, frozen sections and adherent cells (For cells, 6-wells plate or culture dish should be used).
2. Fresh tissue. The immunohistochemistry and immunofluorescence services are provided after the tissue embedding and sectioning.
3. Primary antibodies. Users can offer the primary antibodies by themselves, as well as purchasing from us. Cloud-Clone Corp. can provide more than 10,000 primary antibodies to meet customers' requirements.
4. For double staining, the two primary antibodies customers provided should be derived from different species.

The feedback to the customer

1. Paraffin embedded tissue blocks or sections.
2. Sections used for IHC detection and images.
3. The comprehensive protocol of the experiment.