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CD41 (Platelet Glycoprotein IIb) Antibody Staining Protocol for Immunohistochemistry
Description: CD41 is the α subunit of the CD41/CD61 complex (GPllb-llla), a calcium-dependent, non-covalently associated heterodimer. CD41 is a 135 kDa molecule, consisting of 2 peptides: a 120 kDa α chain and a 23 kDa β chain. The α chain and the β chain are linked by a single disulphide bond. The α chain contains four calcium-binding sites and is entirely extracellular, while the β-chain has extracellular, transmembrane and cytoplasmic domains. The CD41/CD61 complex appears early in megakaryocyte maturation. The activated CD41/CD61 complex is a receptor for von Willebrand factor, soluble fibrinogen and fibronectin and plays a central role in platelet activation and aggregation. CD41 is expressed to a variable degree in megakaryoblastic /cytic leukaemias. It is absent from, or defective on the platelets of patients suffering from Glanzmann's thrombasthenia.
Primary Antibody
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Name: CD41, Platelet Glycoprotein IIb Antibody |
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Clone: 5B12, Mouse anti-Human |
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Supplier: Dako |
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Catalog Number: M7057 |
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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. |
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Incubation Time/Temp: 60 min/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: Cell membrane |
| Images: Search image |
Additional Information:
| Tissue Type: Spleen |
| Fixation: Formalin fixed paraffin sections, or acetone fixed frozen sections |
| Positive Control: Spleen |
| Negative Control: Omit primary antibody, isotype control, 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 on acetone fixed frozen sections |
References: