CNTFR-alpha Antibody Staining Protocol for Immunohistochemistry


Description: Ciliary neurotrophic factor receptor (CNTFR) shares functional and structural properties with members of the hematopoietic cytokine family. Signaling through the CNTFR involves the binding and activation of a multisubunit receptor complex composed of three components: a ligand-specific-α-receptor (CNTFR), signal-transducing β-subunits of gp130 and leukemia inhibitory factor receptor-β (LIFR).1 Binding of CNTF to CNTFR triggers the association of the receptor complex components resulting in activation of a signal transduction cascade mediated by intracellular protein tyrosine kinases. CNTFR can function in a membrane–bound or soluble form; the membrane-bound form is expressed in neuronal and muscle cells, while the soluble form, once processed, can serve as a cofactor to potentiate CNTF actions. This action may serve to enhance the survival, proliferation, and maturation of oligodendrocyte lineage cells.


Primary Antibody

Name: CNTFR alpha Antibody

Clone: AN-H6, Mouse monoclonal

Supplier: BD PharMingen

Catalog Number: 558783

Dilution: 1:100 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: 95-100 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

Reagent: Mayer's Hematoxylin
Staining Time: 30 seconds

Staining Pattern: Membrane
Images: Search image

Additional Information:
Tissue Type: Heart
Fixation: Formalin fixed paraffin sections
Positive Control: Heart
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


Not tested on frozen sections