ACTH (Adrenocorticotrophic Hormone) Antibody Staining Protocol for Immunohistochemistry


Description: Adrenocorticotrophic hormone (ACTH) is a peptide hormone produced by the anterior pituitary gland that stimulates the adrenal cortex. It stimulates the adrenal cortex to secrete glucocorticoid hormones, which help synthesise glucose, catabolise proteins, mobilise free fatty acids and inhibit inflammation in allergic responses. The hormone comprises a long chain of amino acids that share a high degree of homology between humans, cows, pigs and sheep.


Primary Antibody

Name: Adrenocorticotrophic Hormone

Clone: 56, Mouse Anti-Human

Supplier: Novocastra Labs

Catalog Number: NCL-ACTH

Dilution: 1:100 - 1:200 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: Antigen retrieval is not needed
Buffer/pH value: N/A
Heat/Cool Temperature: N/A
Heat/Cool Time: N/A

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: Cytoplasmic
Images: Search image

Additional Information:
Tissue Type: Pituitary gland
Fixation: Formalin fixed paraffin secitons or acetone fixed frozen sections
Positive Control: Pituitary gland
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