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

Name: CD41, Platelet Glycoprotein IIb Antibody

Clone: 5B12, Mouse anti-Human

Supplier: Dako

Catalog Number: M7057

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: 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

Reagent: Mayer's Hematoxylin
Staining Time: 30 seconds

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


Not need to perform antigen retrieval on acetone fixed frozen sections