CD14 Antibody Staining Protocol for Immunohistochemistry

 

Description: CD14 antigen is a GPI-linked glycoprotein with a molecular weight of 55kD. The CD14 antigen is expressed on cells of the myelomonocytic lineage including monocytes, macrophages and Langerhans cells. Low expression is observed on neutrophils and on human B cells. CD14 antigen is a receptor for bacterial lipopolysaccharide (LPS, endotoxin) and the lipopolysaccharide binding protein (LBP). LBP and CD14 antigen serves two physiological roles. These proteins act as opsonin and opsonic receptor, respectively, to promote the phagocytic uptake of bacteria or LPS-coated particles by macrophages.

 

Primary Antibody

Name: CD14 Antibody

Clone: 7, Mouse anti-CD14

Supplier: Novocastra Labs

Catalog Number: NCL-CD14-223

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: IHC-TekTM Epitope Retrieval Steamer Set (Cat# IW-1102)
Buffer/pH value: IHC-TekTM Epitope Retrieval Solution (Cat# IW-1100)
Heat/Cool Temperature: 95-100 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: Membrane/cytoplasmic
Images: Search image

Additional Information:
Tissue Type: Placenta
Fixation: Formalin fixed paraffin sections, not effective on frozen sections
Positive Control: Placenta
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

 

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