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CDX2

IHC of CDX2 on an FFPE Colon Adenocarcinoma Tissue

Description

CDX2 is a caudal-type homeobox gene that encodes an intestine-specific transcription factor expressed early in intestinal development and that may be involved in the regulation of proliferation and differentiation of intestinal epithelial cells. It is expressed in the nuclei of epithelial cells throughout the intestine, from duodenum to rectum.


The CDX2 protein is expressed in Primary and Metastatic Colorectal Carcinomas and has also been demonstrated in the intestinal metaplasia of the stomach and intestinal-type gastric cancer. It is not expressed in the normal gastric mucosa. Loss of CDX2 protein expression has been correlated with loss of differentiation in colorectal cancers. Anti-CDX2 antibody has been useful in distinguishing the gastrointestinal origin of Metastatic Adenocarcinomas and carcinoids. Studies have shown that CDX2 is a superior marker compared to CK20. A high percentage of Mucinous Carcinomas of the Ovary also stain positively with this antibody, as well as Carcinomas from the upper gastrointestinal tract.

Antibody Type
Rabbit Monoclonal
Clone
EPR2764Y
Isotype
IgG
Reactivity
Paraffin, Frozen
Localization
Nuclear
Control
Adenocarcinoma of Colon, Normal Colon
Storage
Store at 2°-8°C
Stability
Up to 36 Months

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

Anti-CDX2 is a rabbit monoclonal antibody derived from cell culture supernatant that is concentrated, dialyzed, 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 6057
Prediluted
Ready-To-Use
3.0 ml
BSB 6058
Prediluted
Ready-To-Use
7.0 ml
BSB 6059
Prediluted
Ready-To-Use
15.0 ml
BSB 6060
Concentrated
1:10-1:50
0.1 ml
BSB 6061
Concentrated
1:10-1:50
0.5 ml
BSB 6062
Concentrated
1:10-1:50
1.0 ml
BSB 6063
Control Slides
 
5
Note: For concentrated antibodies, please centrifuge prior to use to ensure recovery of all product.
References
  1. Levine PH, et al. Diagn Cytopathology. 2006;Mar;34(3):191-5
  2. Mazziotta RM, et al. App Immunohistochem Mol Morphol. 2005;Mar;13(1):55-60
  3. Saqi A, et al. Am J Clin Pathol. 2005;Mar;123(3):394-404
  4. Erickson LA, et al. Endocr Pathol. 2004;fall;15(3):247-52
  5. Saad RS, et al. AM J Clin Pathol. 2004;Sep;122(3):421-7
  6. Kaimaktchiev V, et al. Mod Pathol. 2004;Nov;17(11):1392-9

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