Mouse Cytokeratin 20 Monoclonal Antibody | anti-KRT20 antibody

Cytokeratin 20

Cat. Num. AAA370051

• Reactivity: Human, Dog, Cat, Mouse
• Applications: Immunohistochemistry
• Synonyms: Cytokeratin 20; Monoclonal Antibody; anti-KRT20 antibody

• Host: Mouse
• Reactivity: Human, Dog, Cat, Mouse
• Clonality: Monoclonal
• Isotype: IgG2a/K
• Clone Number: Ks20.8
• Form/Format: Cytokeratin 20 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.

• Applicable Applications for anti-KRT20 antibody: Immunohistochemistry (IHC)
• Application Notes: This antibody is intended for use in Immunohistochemical applications on formalin-fixed paraffin-embedded tissues (FFPE), frozen tissue sections and cell preparations.; ; Specimen Preparation:; Paraffin sections: The antibody can be used on formalin-fixed paraffin-embedded (FFPE) tissue sections. Ensure tissue undergoes appropriate fixation for best results. Pre-treatment of tissues with heat-induced epitope retrieval (HIER) is recommended using ImmunoDNA Retriever with Citrate , ImmunoDNA Retriever with EDTA or ImmunoDNA Digestor. See reverse side for complete protocol. Tissue should remain hydrated via use of Immuno/DNA Washer solutions.; Frozen sections and cell preparations: The antibody can be used for labeling acetone-fixed frozen sections and acetone-fixed cell preparations.; ; Staining Procedure:; 1. Cut and mount 3-5 micron formalin-fixed paraffin-embedded tissues on positively charged slides such as Hydrophilic Plus Slides.; 2. Air dry for 2 hours at 58°C.; 3. Deparaffinize, dehydrate and rehydrate tissues.; 4. Subject tissues to heat induced epitope retrieval (HIER) using a suitable retrieval solution such as ImmunoDNA Retriever with Citrate or EDTA.; 5. Any of three heating methods may be used:; a. TintoRetriever Pressure Cooker or Equivalent; Place tissues/slides in a staining dish or coplin jar containing the ImmunoDNA Retriever with Citrate or EDTA, and place on trivet in the pressure cooker. Add 1-2 inches of distilled water to the pressure cooker and turn heat to high. Incubate for 15 minutes. Open and immediately transfer slides to room temperature.; b. TintoRetriever PT Module or Water Bath Method; Place tissues/slides in a pre-warmed staining dish or coplin jar containing the ImmunoDNA Retriever with Citrate or EDTA at 95°-99°C. Incubate for 30-60 minutes.; c. 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. For manual staining, perform antibody incubation at ambient temperature. For automated staining methods, perform antibody incubation according to instrument manufacturer’s instructions.; 8. Wash slides with ImmunoDNA washer or DI water.; 9. Continue IHC staining protocol. Wash slides between each step with ImmunoDNA washer solution.; ; Product Limitations: Due to inherent variability present in immunohistochemical procedures (including fixation time of tissues, dilution factor of antibody, retrieval method utilized and incubation time), optimal performance should be established through the use of positive and negative controls.
• Immunogen: Semi-purified human cytokeratin preparation.
• Control: Colon Carcinoma, Colon Mucosa, Bladder
• Localization: Cytoplasmic
• Reactivity Note: Paraffin, Frozen
• Precautions: 1. This product contains <0.1% sodium azide (NaN3) as a preservative. Ensure proper handling procedures are used with this reagent.; 2. Always wear personal protective equipment such as laboratory coat, goggles and gloves when handling reagents.; 3. Dispose of unused solution with copious amount of water.; 4. Dispose of unused solution with copious amount of water.; 5. Avoid contact with eyes. If contact occurs, flush with large quantities of water.; 6. Follow safety precautions of the heating device used for epitope retrieval (TintoRetriever Pressure Cooker or similar).; 7. For additional safety information refer to Safety Data Sheet for this product.; 8. For complete recommendations for handling biological specimens, please refer to the CDC document, “Guidelines for Safe Work Practices in Human and Animal Medical (see References in this document).
• Mounting Protocols: For detailed instructions using biodegradable permanent mounting media such as XyGreen PermaMounter or organic solvent based resin such as PermaMounter, please inquire.
• Preparation and Storage: Store at 2-8°C (Control Slides: Store at 20-25°C)

Dilution Information

Immunohistochemistry (IHC)

(IHC of Cytokeratin 20 on an FFPE Adenocarcinoma Tissue)

Abbreviated Immunohistochemical Protocol

Performance Characteristics

Related Product Information for anti-KRT20 antibody

Cytokeratin 20 (CK20) is a 46 kDa intermediate filament protein whose expression is restricted primarily to gastric and intestinal epithelium, urothelium, and Merkel cells. Cytokeratin 20 is a Type I cytokeratin. It is a major cellular protein of mature enterocytes and goblet cells found in the gastric and intestinal mucosa. CK 20 is expressed in Adenocarcinomas of the colon, stomach, pancreas and biliary system. It is also expressed in Mucinous Ovarian Tumors, Transitional-cell Carcinomas of the urinary tract, and Merkel-cell Carcinomas. Cytokeratin 20 is useful in the differentiation of specific types of simple epithelial cells of the urinary tract and normal and malignantly-transformed epithelia. This antibody is essentially non-reactive in Squamous Cell Carcinomas and Adenocarcinomas of the Breast, Lung, and Endometrium, Non-mucinous Tumors of the Ovary, and Small-cell Carcinomas. This antibody is often used in conjunction with CK7 and other antibodies to distinguish Colon Carcinomas (CK20+) from Ovarian, Pulmonary, and Breast Carcinomas.

NCBI and Uniprot Product Information

• NCBI GI #: 402645
• UniProt Accession #: P35900; B2R6W7
• NCBI Official Full Name: cytokeratin 20
• UniProt Protein Name: Keratin, type I cytoskeletal 20
• Protein Family: Keratin
• UniProt Gene Name: KRT20
• UniProt Entry Name: K1C20_HUMAN

Uniprot Description

Function: Plays a significant role in maintaining keratin filament organization in intestinal epithelia. When phosphorylated, plays a role in the secretion of mucin in the small intestine

By similarity. Ref.4 Ref.8

Subunit structure: Heterotetramer of two type I and two type II keratins. Associates with KRT8.

Subcellular location: Cytoplasm Ref.6 Ref.7.

Tissue specificity: Expressed predominantly in the intestinal epithelium. Expressed in luminal cells of colonic mucosa. Also expressed in the Merkel cells of keratinized oral mucosa; specifically at the tips of some rete ridges of the gingival mucosa, in the basal layer of the palatal mucosa and in the taste buds of lingual mucosa. Ref.6 Ref.7

Developmental stage: First detected at embryonic week 8 in individual 'converted' simple epithelial cells of the developing intestinal mucosa. In later fetal stages, synthesis extends over most goblet cells and a variable number of villus enterocytes. In the developing gastric and intestinal mucosa, expressed in all enterocytes and goblet cells as well as certain 'low-differentiated' columnar cells, whereas the neuroendocrine and Paneth cells are negative. Ref.1

Post-translational modification: Hyperphosphorylation at Ser-13 occurs during the early stages of apoptosis but becomes less prominent during the later stages. Phosphorylation at Ser-13 also increases in response to stress brought on by cell injury

By similarity.Proteolytically cleaved by caspases during apoptosis. Cleavage occurs at Asp-228. Ref.4

Miscellaneous: There are two types of cytoskeletal and microfibrillar keratin: I (acidic; 40-55 kDa) and II (neutral to basic; 56-70 kDa).

Sequence similarities: Belongs to the intermediate filament family.