Immunohistochemistry (IHC)
(Colorectal adenocarcinoma with diffuse strong expression of CK20 (A and B) and colorectal adenocarcinoma and normal epithelium of the large intestine mucosa with expression
of CK20 (C). Adenoma of the large intestine with expression of CK20 (D). All, formalin fixed, paraffin embedded human tissues (4 um sections) stained with anti - Cytokeratin 20 (MBS684193) monospecific clonal antibody according to related datasheet.)
Rabbit Cytokeratin 20 Monoclonal Antibody | anti-KRT20 antibody
Anti - Cytokeratin 20
IHC-P PROTOCOL – INSTRUCTION MANUAL
1. Deparaffinize the section in 3 changes of xylene, 5 minutes each.
2. Wash the section in 96%, 80% and 70% ethyl alcohol for 10 minutes each.
3. Rinse in distilled water.
4. Block the endogenous peroxidase by incubating the tissue in 3% hydrogen peroxide (H2O2) for 10 minutes.
5. Wash in distilled water.
6. For antigen retrieval: immerse the slide in Tris-EDTA buffer, pH 9.0, 0.05% Tween-20* and incubate at 95ºC in water bath for 60 minutes.
7. Remove the staining to room temperature and let the slide to cool (in Tris-EDTA buffer, pH 9.0) for 15 minutes.
8. Rinse in distilled water.
9. Wash in 0.05 M Tris-HCl, pH 7.6 buffer supplemented with 0.2% of Tween-20 (buffer A) for 5 minutes.
10. CONCENTRATED:
Incubate the section with primary antibody at the dilution 1:100 - 1:200 for 1 hour in the closed wet chamber.
READY TO USE (RTU):
Incubate the section with primary antibody (ready to use) for 1 hour in a closed wet chamber.
11. Wash twice 5 minutes with buffer A.
12. Apply the secondary antibody (the protocol depends on the supplier), and proceed to standard immunohistochemistry protocol (HRP - Peroxide - DAB). Micropolymer-HRP detection kit rabbit/mouse dual is suggested (Please Inquire).
13. Wash twice 5 minutes with buffer A.
14. Apply the chromogen (DAB), 1 - 3 minutes.
15. Wash in water – 10 minutes.
16. Stain in hematoxylin for 5 minutes.
17. Wash in water – 10 minutes.
18. Dehydrate the section in 2 changes of 96% ethyl alcohol for 5 minutes each.
19. Wash the section in 2 changes of xylene for 2 minutes each.
20. Mount the slide for observation.
* Tris-EDTA Buffer (10mM Tris Base, 1mM EDTA solution, 0.05% Tween-20, pH 9.0):
Tris ------------- 1.21 g; EDTA ------------ 0.37 g; Distilled water -------------------------- 1000 ml
Mix to dissolve in 700 ml of distilled water. Adjust pH to 9.0 with 1M HCl and then add 0.5 ml of Tween-20 and mix well. Adjust the final volume to 1 liter with distilled water. Store this solution at room temperature for 3 months or at +4ºC for longer storage.
2. Centrifuge the vial before use.
3. Intended for use in laboratories.
4. Do not use after expiration date stamped on vial label.
5. Avoid contamination of the reagent.
6. Any discrepancies in the recommended procedures stated in the working protocol may affect the final results.
7. The reagent contains sodium azide (NaN3) which is highly toxic in higher concentrations. The concentration in the reagent (0.05%) is not considered as hazardous.
8. Disposal of waste material must be conducted in accordance with local regulations.
9. Wear appropriate Personal Protective Equipment to avoid contact with eyes and skin.
Immunohistochemistry (IHC)
(Colorectal adenocarcinoma with diffuse strong expression of CK20 (A and B) and colorectal adenocarcinoma and normal epithelium of the large intestine mucosa with expression of CK20 (C). Adenoma of the large intestine with expression of CK20 (D). All, formalin fixed, paraffin embedded human tissues (4 um sections) stained with anti - Cytokeratin 20 (MBS684193) monospecific clonal antibody according to related datasheet.)
Immunohistochemistry (IHC)
(Colorectal adenocarcinoma with diffuse strong expression of CK20 (A and B) and colorectal adenocarcinoma and normal epithelium of the large intestine mucosa with expression
of CK20 (C). Adenoma of the large intestine with expression of CK20 (D). All, formalin fixed, paraffin embedded human tissues (4 um sections) stained with anti - Cytokeratin 20 (MBS684193) monospecific clonal antibody according to related datasheet.)
NCBI and Uniprot Product Information
NCBI Description
The protein encoded by this gene is a member of the keratin family. The keratins are intermediate filament proteins responsible for the structural integrity of epithelial cells and are subdivided into cytokeratins and hair keratins. The type I cytokeratins consist of acidic proteins which are arranged in pairs of heterotypic keratin chains. This cytokeratin is a major cellular protein of mature enterocytes and goblet cells and is specifically expressed in the gastric and intestinal mucosa. The type I cytokeratin genes are clustered in a region of chromosome 17q12-q21. [provided by RefSeq, Jul 2008]
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.
Research Articles on KRT20
Similar Products
Product Notes
The KRT20 krt20 (Catalog #AAA684193) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. AAA Biotech's Cytokeratin 20 can be used in a range of immunoassay formats including, but not limited to, Immunohistochemistry (IHC) - Paraffin. IHC-P READY-TO-USE IHC-P PROTOCOL – INSTRUCTION MANUAL 1. Deparaffinize the section in 3 changes of xylene, 5 minutes each. 2. Wash the section in 96%, 80% and 70% ethyl alcohol for 10 minutes each. 3. Rinse in distilled water. 4. Block the endogenous peroxidase by incubating the tissue in 3% hydrogen peroxide (H2O2) for 10 minutes. 5. Wash in distilled water. 6. For antigen retrieval: immerse the slide in Tris-EDTA buffer, pH 9.0, 0.05% Tween-20* and incubate at 95ºC in water bath for 60 minutes. 7. Remove the staining to room temperature and let the slide to cool (in Tris-EDTA buffer, pH 9.0) for 15 minutes. 8. Rinse in distilled water. 9. Wash in 0.05 M Tris-HCl, pH 7.6 buffer supplemented with 0.2% of Tween-20 (buffer A) for 5 minutes. 10. CONCENTRATED: Incubate the section with primary antibody at the dilution 1:100 - 1:200 for 1 hour in the closed wet chamber. READY TO USE (RTU): Incubate the section with primary antibody (ready to use) for 1 hour in a closed wet chamber. 11. Wash twice 5 minutes with buffer A. 12. Apply the secondary antibody (the protocol depends on the supplier), and proceed to standard immunohistochemistry protocol (HRP - Peroxide - DAB). Micropolymer-HRP detection kit rabbit/mouse dual is suggested (Please Inquire). 13. Wash twice 5 minutes with buffer A. 14. Apply the chromogen (DAB), 1 - 3 minutes. 15. Wash in water – 10 minutes. 16. Stain in hematoxylin for 5 minutes. 17. Wash in water – 10 minutes. 18. Dehydrate the section in 2 changes of 96% ethyl alcohol for 5 minutes each. 19. Wash the section in 2 changes of xylene for 2 minutes each. 20. Mount the slide for observation. * Tris-EDTA Buffer (10mM Tris Base, 1mM EDTA solution, 0.05% Tween-20, pH 9.0): Tris ------------- 1.21 g; EDTA ------------ 0.37 g; Distilled water -------------------------- 1000 ml Mix to dissolve in 700 ml of distilled water. Adjust pH to 9.0 with 1M HCl and then add 0.5 ml of Tween-20 and mix well. Adjust the final volume to 1 liter with distilled water. Store this solution at room temperature for 3 months or at +4ºC for longer storage. Researchers should empirically determine the suitability of the KRT20 krt20 for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "Cytokeratin 20, Monoclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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