 |
|
GFAP Antibody Staining Protocol for Immunohistochemistry
Description: Glial fibrillary acidic protein (GFAP) is an intermediate filament protein of 52kD found in glial cells such as astrocytes and ependymal cells. In the peripheral nervous system, GFAP has been demonstrated in Schwann cells, enteric glial cells and satellite cells of human sensory ganglia. In neoplastic tissues, this antibody is useful for the identification of astrocytomas and ependymomas.
Primary Antibody
|
Name: GFAP Antibody |
|
Clone: Rabbit anti-Cow |
|
Supplier: Dako |
|
Catalog Number: Z0334 |
|
Dilution: 1:500 using IHC-TekTM Antibody Diluent (Cat# IW-1000 or IW-1001) to reduce background and unspecific staining and serum blocking step is NOT needed. |
|
Incubation Time/Temp: 60 minutes/room temperature |
Antigen Retrieval
| Device: IHC-TekTM Epitope Retrieval Steamer Set (Cat# IW-1102) |
| Buffer/pH value: IHC-TekTM Epitope Retrieval Solution (Cat# IW-1100) |
| Heat/Cool Temperature: 95 ºC - 100 ºC/room temperature |
| Heat/Cool Time: 20 minutes/20 minutes |
Detection Methods
| Standard Method: ABC Method or LSAB Method |
| Enhanced Method: Polymeric Methods |
Chromogen Substrate
| Reagent: DAB |
| Incubation Time/Temperature: 1-3 minutes/room temperature |
Counterstain
| Reagent: Mayer's Hematoxylin |
| Staining Time: 30 seconds |
Results:
| Staining Pattern: Cytoplasmic (glial cells such as astrocytes and ependymal cells) |
| Images: Search image |
Additional Information:
| Tissue Type: Brain (cerebellum, striatum) |
| Fixation: Formalin fixed paraffin sections, or acetone fixed frozen sections |
| Positive Control: Brain (cerebellum, striatum) |
| Negative Control: Omit primary antibody, isotype control or absorption control |
| Blocking: 2-5% normal serum to reduce unspecific background staining; 0.5-3% H2O2 to block endogenous peroxidase activity; avidin/biotin to block endogenous biotin activity if necessary |
Notes:
No need to perform antigen retrieval on frozen sections |
References: