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Cytokeratin 17

IHC of CK 17 on an FFPE Cervical Cancer Tissue

Description

Cytokeratin 17 is a Type I cytokeratin with a MW of 46 kD found sometimes in association with Cytokeratin 7. It is found in nail beds, hair follicles, sebaceous glands, and other epidermal appendages. Mutations in the gene encoding this protein lead to Jackson-Lawler type Pachyonychia Congenita and Steatocystoma Multiplex.


Cytokeratin 17 antibody has been used to distinguish immature Cervical Squamous Metaplasia from high grade Cervical Intraepithelial Neoplasia (CIN III). Anti-CK 17 also labels myoepithelial cells in the benign breast tissue. CK 17 labeling of Breast Carcinoma cells (so-called basal phenotype) has been associated with a poor prognosis.

Antibody Type
Mouse Monoclonal
Clone
E3
Isotype
IgG2b
Reactivity
Paraffin, Frozen
Localization
Cytoplasmic
Control
Small Intestine, Colon Mucosa, Bladder, Thyroid Carcinoma
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

Cytokeratin 17 is a mouse 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 6184
Prediluted
Ready-To-Use
3.0 ml
BSB 6185
Prediluted
Ready-To-Use
7.0 ml
BSB 6186
Prediluted
Ready-To-Use
15.0 ml
BSB 6187
Concentrated
1:50-1:200
0.1 ml
BSB 6188
Concentrated
1:50-1:200
0.5 ml
BSB 6189
Concentrated
1:50-1:200
1.0 ml
BSB 6190
Control Slides
 
5
Note: For concentrated antibodies, please centrifuge prior to use to ensure recovery of all product.
References
  1. Smith FJ, et al. J. Invest. Dermatol. 2001;116(5):806-8
  2. Van de Rijn M, et al. Am. J. Pathol. 2003;161(6):1991-6
  3. Regauer S, Histopathology. 2007;Apr;50(5):629-35
  4. Chu PG, et al. Am J Surg Pathol. 2005;Mar;29(3):359-67
  5. Martens JE, et al. Anticancer Res. 2004;Mar-Apr;24(2B):771-5

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