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Cytokeratin 14

IHC of CK14 on an FFPE Cervical Tissue

Description

Cytokeratin 14 is a Type I polypeptide found in basal cells of squamous epithelia, some glandular epithelia, myoepithelium, and mesothelial cells. It is usually found as a heterotetramer with two cytokeratin 5 molecules, and a Type II keratin. Together, they form the cytoskeleton of epithelial cells. Mutations in the genes for these cytokeratins are associated with Epidermolysis Bullosa Simplex.


Cytokeratin 14 has been studied as a prognostic marker in Breast Cancer. This antibody labels the basal layer of stratifying squamous and non-squamous epithelia. The staining pattern is cytoplasmic. It recognizes Basal Cell Carcinomas and Squamous Cell Carcinomas. Anti-CK14 has been demonstrated to be useful in differentiating Squamous Cell Carcinomas from other epithelial tumors. This antibody has also been useful in separating oncocytic tumors of the kidney from renal mimics, as well as in determining metaplastic Carcinomas of the Breast.

Antibody Type
Mouse Monoclonal
Clone
LL002
Isotype
IgG3
Reactivity
Paraffin, Frozen
Localization
Cytoplasmic
Control
Squamous Mucosa, Squamous 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

Anti-Cytokeratin 14 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 6219
Prediluted
Ready-To-Use
3.0 ml
BSB 6220
Prediluted
Ready-To-Use
7.0 ml
BSB 6221
Prediluted
Ready-To-Use
15.0 ml
BSB 6222
Concentrated
1:50-1:200
0.1 ml
BSB 6223
Concentrated
1:50-1:200
0.5 ml
BSB 6224
Concentrated
1:50-1:200
1.0 ml
BSB 6225
Control Slides
 
5
Note: For concentrated antibodies, please centrifuge prior to use to ensure recovery of all product.
References
  1. Reis-Filho JS, et al. Appl Immunohistochem Mol Morphol. 2003;Mar;11(1):1-8
  2. Chu PG, et al. Histopathology. 2001;Jul;39(1):9-16
  3. Chu PG, Weiss LM. Histopathology. 2001;Nov;39(5):455-62
  4. Dabbs David J, Diagnostic Immunohistochemistry. Churchill-Livingstone. 2002;166-176

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