 |
|
CD31 Antibody Staining Protocol for Immunohistochemistry
Description: This antibody reacts with rat Pecam-1. Antibody gives positive reaction at high dilutions against recombinant rat PECAM-1. This antibody also cross reacts with Pig endothelial cells and platelets as determined by histochemistry on acetone fixed frozen sections and flow cytometry. Reactivity with other species not determined. Non reactive with human CD31.
Primary Antibody
|
Name: CD31 Antibody |
|
Clone: TLD-3A12, Mouse anti-Rat |
|
Supplier: Research Diagnostics |
|
Catalog Number: RDI-RTCD31-3A |
|
Dilution: 1:50 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: 60min/room temperature |
Antigen Retrieval
| Device: Incubator |
| Buffer/pH value: IHC-TekTM Proteinase K Solution (Cat# IW-1101) |
| Heat/Cool Temperature: 37 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 of endothelial cells and platelets |
| Images: Search image |
Additional Information:
| Tissue Type: Tonsil |
| Fixation: Zinc fixed paraffin sections, or acetone fixed frozen sections |
| Positive Control: Tonsil, cardiac tissues, skin, kidney |
| 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:
|
Not need to perform antigen retrieval for acetone fixed frozen sections |
References: