Immunohistochemistry (IHC) refers to the method of localizing specific antigens in cells or tissue sections based on the binding of labeled antibody to antigens. The antigen-antibody interaction could be visualized by a marker such as enzyme, radiolabel, fluorophore or colloidal metal. IHC makes it possible to visualize the distribution and localization of specific cellular components within a cell or tissue.
There are two methods for the immunohistochemical detection of antigens in tissue, the direct and indirect methods. The direct method, it is one-step staining with a labeled antibody reacting directly to antigen in tissue sections. This method is simple and rapid but lack of sensitivity. The indirect method requires the use of two antibodies. One is unlabeled primary antibody which reacts with tissue antigen and the other is a labeled secondary antibody which react with this primary antibody. The indirect method is more sensitive due to several secondary antibody reactions with different antigenic sites on the primary antibody and allows for signal amplification. Abnova uses the indirect method to detect antigens in tissue.
10 tissue microarray slides for Alzheimer disease and cancer research.
Abnova offers a complete immunohistochemistry service for researchers. All studies are conducted under R&D conditions and the results are proprietary to the customer.
General Workflow of IHC Service
Customer completes a IHC custom service form (PDF, Excel File) and send to Abnova.
Abnova prepares slides from formalin-fixed and paraffin-embedded tissues.
Abnova performs IHC staining either on slides without pretreatment or with pretreatment, or both.
Abnova performs dilutions of antibody according customer's instruction or performs a serial dilution of 40, 20, 10, 5, 2.5,1.25, 0.675 ug/ml.
Abnova offers the following slide pretreatments. Customer may choose one or more protocols for the pretreatment:
Pretreatment in 1X citrate buffer (pH 6.0), microwave at 750W for 20 minutes, and subsequently cool the samples.
Pretreatment in 1X Tris/EDTA buffer (pH 9.0), microwave at 750W for 20 minutes, and subsequently cool the samples.
Pretreatment in HCl (2N) (pH 0.6~0.9) at room temperature for 10~20 minutes.
Pretreatment in 0.1% trypsin and shake for 25 minutes at 37C.
Immunohistochemistry is a method of detecting the presence of specific proteins in cells or tissues. It is widely used in basic research to understand the distribution and localization of biomarkers and differentially expressed proteins in different parts of a biological tissue.
Rotary microtome is an instrument used to cut the blocks of tissue embedded in paraffin into micro fine slices (known as sections). Microtome is an important device in microscopy preparation, allowing for the preparation of samples for observation under transmitted light or electron radiation.