Chromogranin A (CGA) Antibody Staining Protocol for Immunohistochemistry


Description: Chromogranin A is an acidic, soluble protein first identified in adrenal chromoffin granules. Immunohistochemical studies have demonstrated chromogranin A in most granule-containing endocrine cells, central and peripheral nerves, as well as in most neuroendoctrine tumors. This antibody reacts with human chromagranin A, but not with fragments located on the N-terminal half of the protein. Traces of antibodies to other human plasma proteins and human urine proteins have been removed by solid phase absorption. The antibody reacts with chromogranin A in neuroendocrine cells of the human stomach, colon, pancreas, adrenal gland, peripheral nerve tissue and carcinoid tumors.


Primary Antibody

Name: Chromogranin A Antibody

Clone: Polyclonal, Rabbit anti-Human

Supplier: Dako

Catalog Number: A0430

Dilution: 1:1500 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: Cytoplasmic
Images: Search image

Additional Information:
Tissue Type: Stomach, colon, pancreas, adrenal gland, peripheral nerve
Fixation: Formalin fixed paraffin sections
Positive Control: Stomach, colon, pancreas, adrenal gland, peripheral nerve
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


1. CGA (Clone LK2H10, Boehringer Mannhein, Cat# 1199021) and (Clone LK2H10+PHE5, Lab Vision, Cat MS-382-P) also works well at 1:5000 with Tris-EDTA pretreatment and 1:3000 with citrate buffer pretreatment, respectively.