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

Counterstain

Reagent: Mayer's Hematoxylin

Staining Time: 30 seconds

Results:

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

 

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