HomeProductsDistributorsContact UsCareers

Back to Antibodies for Immunohistochemistry

PDF

ACTH

IHC of ACTH on an FFPE Pituitary Tissue

Description

Adrenocorticotropic Hormone (ACTH or corticotropin) is a polypeptide hormone synthesized from POMC, (Pro-opiomelanocortin) and secreted from corticotropes in the anterior lobe of the pituitary gland in response to Corticotropin-releasing Hormone (CRH) released by the hypothalamus. It consists of 39 amino acids.


ACTH is a useful marker in the classification of pituitary tumors and the study of pituitary disease. It reacts with ACTH-producing cells (corticotrophs), as well as other tumors (e.g.,some Small-Cell Carcinomas present in lung tissue) causing Paraneoplastic Syndromes by secreting ACTH.

Antibody Type
Rabbit Ployclonal
Clone
N/A
Isotype
N/A
Reactivity
Paraffn, Frozen
Localization
Cytoplasmic
Control
Normal Pituitary
Storage
Store at 2°-8°C
Stability
2 years

For long-term storage of the concentrated antibody, it is recommended that aliquots of the antibody be frozen at -20°C in glycerol 50% (frost-free freezers are not recommended). Repeated freezing and thawing must be avoided. Dilute using an antibody diluent such as ImmunoDetector Protein Block/Antibody Diluent (BSB 0040 and BSB 0041) or ImmunoDNA Background Blocker (BSB 0103-BSB 0107).

Presentation

ACTH is a purifified immunoglobulin fraction of rabbit antiserum that is filter sterilized and diluted in buffer pH 7.5, containing BSA and sodium azide as a preservative.

Availability
Catalog No.
Antibody Type
Dilution
Volume/QTY
BSB 5015
Prediluted
Ready-To-Use
3.0 ml
BSB 5016
Prediluted
Ready-To-Use
7.0 ml
BSB 5017
Prediluted
Ready-To-Use
15.0 ml
BSB 5018
Concentrated
1:500-1:2000
0.1 ml
BSB 5019
Concentrated
1:500-1:2000
0.5 ml
BSB 5020
Concentrated
1:500-1:2000
1.0 ml
BSB 5021
Control Slides
 
5
Note: For concentrated antibodies, please centrifuge prior to use to ensure recovery of all product.
References
  1. Hassoun J, et al. Arch Pathol Lab Med. 1982;106:25
  2. Mukai K, et al. 1980;42:42
  3. Kovacs K, Rosai J, New York: Masson Publ. 1980;3

Recommended Immunohistochemical Protocol

Pretreatment
  1. Cut and mount 3-4 micron formalin-fixed paraffin-embedded tissues on positive charged slides.
  2. Air dry for 2 hours at 58° C.
  3. Deparaffinize, dehydrate and rehydrate tissues.
  4. Subject tissues to heat epitope retrieval using a suitable retrieval solution such as ImmunoDNA Retriever with Citrate (BSB 0020-BSB 0023) or EDTA (BSB 0030-BSB 0033).
  5. Any of three heating methods may be used:
    • Electric Pressure Cooker
      Place standoff rack at base of pressure cooker. Add 1-2 inches of distilled water to the pressure cooker and turn heat to high, and incubate for 15 minutes. Open and immediately transfer slides to room temperature.
    • Water Bath Method
      Place tissues/slides in a pre-warmed staining dish or coplin jar containing the ImmunoDNA Retriever with Citrate or EDTA in a water bath set at 95°-99° C. Incubate for 30-60 minutes.
    • Conventional Steamer Method
      Place tissues/slides in a pre-warmed staining dish or coplin jar containing the ImmunoDNA Retriever with Citrate or EDTA in a Steamer, cover and steam for 30-60 minutes.
  6. After heat treatment, transfer slides in ImmunoDNA Retriever with Citrate or EDTA to room temperature and let stand for 15-20 minutes.
  7. Wash slides with IHC wash buffer or DI water.
  8. Continue IHC staining protocol.

Immunohistochemical Protocol

Step

ImmunoDetector
(AP or HRP)

PolyDetector
(AP or HRP)
Peroxidase/AP Block
5 minutes
5 minutes
Primary Antibody
30 minutes
45 minutes
Secondary Biotinylated Link
10 minutes
Not Applicable
AP or HRP Label
10 minutes
45 minutes
Substrate-Chromogen
5-10 minutes
10 minutes
Counterstaining
Time varies with counterstain
Time varies with counterstain