Immunohistochemistry (IHC)
Rabbit Cytokeratin 5/6 Antibody | anti-KRT5 antibody
Anti-Cytokeratin 5/6
Gene Names
KRT5; K5; CK5; DDD; DDD1; EBS2; KRT5A
Applications
Immunohistochemistry
Synonyms
Cytokeratin 5/6; Anti-Cytokeratin 5/6; anti-KRT5 antibody
Host
Rabbit
Clone Number
T16-K
Specificity
Human
Applicable Applications for anti-KRT5 antibody
Immunohistochemistry-Paraffin (IHC-P)
Application Notes
Dilution: 1:100 - 1:200
IHC-P Protocol - Instruction Manual:
1. Deparaffinize the section in 3 changes of xylene, 10 minutes each.
2. Wash the section in 96%, 80% and 70% ethyl alcohol for 10 minutes each.
3. Rinse in distilled water, 2 x 5 minutes.
4. Block the endogenous peroxidase by incubating the tissue in 3% hydrogen peroxide (H2O2) for 10 minutes.
5. Wash in distilled water, 2 x 5 minutes.
6. For antigen retrieval: Immerse the slide in Tris-EDTA buffer, pH 9.0 and incubate at 95-97ºC in water bath for 25 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, 2 x 5 minutes.
9. Wash in PBS (phosphate buffer saline, pH 7.0-7.5) supplemented with 0.05% of Tween-20 (buffer A), 2 x 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.
11. Wash 3 x 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).
13. Wash 3 x 5 minutes with buffer A.
14. Apply the chromogen (DAB), 1 - 3 minutes.
15. Wash in water, 2 x 5 minutes.
16. Stain in hematoxylin for 5 minutes.
17. Wash in distilled water, 3 x 2 minutes.
18. 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.
Ventana Protocol - Instruction Manual:
(Short Application Protocol for Ventana Benchmark Slide Staining System)
1. Deparafinization (Enter).
2. Heating glass (72°C) with the medium temperatures. Deparafinization.
3. Prolonged deparafinization (Enter).
4. Cell conditioning (Enter).
5. ULTRA Conditioner #1 (Enter).
6. Heating glass (96°C), incubation 8 min (Cell conditioner #1).
7. ULTRA CC1 solution application – 20 min (Enter).
8. Titration (Enter).
9. Hand apply - primary antibody. Incubation 32 min.
10. UltraWash (Enter).
11. Nuclear stain (Enter).
12. Hematoxylin application - one drop (nuclear stain), cover and incubate 8 min.
13. After nuclear stain (Enter).
14. Bluing reagent application, one drop. After nuclear stain, cover and incubate 4 min.
IHC-P Protocol - Instruction Manual:
1. Deparaffinize the section in 3 changes of xylene, 10 minutes each.
2. Wash the section in 96%, 80% and 70% ethyl alcohol for 10 minutes each.
3. Rinse in distilled water, 2 x 5 minutes.
4. Block the endogenous peroxidase by incubating the tissue in 3% hydrogen peroxide (H2O2) for 10 minutes.
5. Wash in distilled water, 2 x 5 minutes.
6. For antigen retrieval: Immerse the slide in Tris-EDTA buffer, pH 9.0 and incubate at 95-97ºC in water bath for 25 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, 2 x 5 minutes.
9. Wash in PBS (phosphate buffer saline, pH 7.0-7.5) supplemented with 0.05% of Tween-20 (buffer A), 2 x 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.
11. Wash 3 x 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).
13. Wash 3 x 5 minutes with buffer A.
14. Apply the chromogen (DAB), 1 - 3 minutes.
15. Wash in water, 2 x 5 minutes.
16. Stain in hematoxylin for 5 minutes.
17. Wash in distilled water, 3 x 2 minutes.
18. 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.
Ventana Protocol - Instruction Manual:
(Short Application Protocol for Ventana Benchmark Slide Staining System)
1. Deparafinization (Enter).
2. Heating glass (72°C) with the medium temperatures. Deparafinization.
3. Prolonged deparafinization (Enter).
4. Cell conditioning (Enter).
5. ULTRA Conditioner #1 (Enter).
6. Heating glass (96°C), incubation 8 min (Cell conditioner #1).
7. ULTRA CC1 solution application – 20 min (Enter).
8. Titration (Enter).
9. Hand apply - primary antibody. Incubation 32 min.
10. UltraWash (Enter).
11. Nuclear stain (Enter).
12. Hematoxylin application - one drop (nuclear stain), cover and incubate 8 min.
13. After nuclear stain (Enter).
14. Bluing reagent application, one drop. After nuclear stain, cover and incubate 4 min.
Buffer
20 mM Tris-HCl, pH 8.0
Stabilizer
20 mg/ml BSA
Preservative
0.05% NaN3
Immunogen
Peptide derived from C-terminal sequence of human cytokeratin 6. Antibody recognizes the epitope (identical with cytokeratin 5) between Thr548 – His564.
Cellular Localization
Cytoplasm
Positive Control
Skin or prostate tissue (basal cells); squamous cell carcinoma tissue
Precautions
1. We strongly recommend to use Primary Antibody Diluent, eventually alternative diluent (containing protease free BSA at the concentrations > 1mg/ml) for dilution of concentrated antibodies, otherwise the warranty might be voided.
2. Centrifuge the vial before use.
3. Intended for professional 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.
2. Centrifuge the vial before use.
3. Intended for professional 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.
Preparation and Storage
Store at +4°C
Immunohistochemistry (IHC)
Immunohistochemistry (IHC)
NCBI and Uniprot Product Information
NCBI GI #
NCBI GeneID
NCBI Accession #
NCBI GenBank Nucleotide #
NCBI Official Full Name
keratin, type II cytoskeletal 5
NCBI Official Synonym Full Names
keratin 5
NCBI Official Symbol
KRT5
NCBI Official Synonym Symbols
K5; CK5; DDD; DDD1; EBS2; KRT5A
NCBI Protein Information
keratin, type II cytoskeletal 5
UniProt Protein Name
Keratin, type II cytoskeletal 5
Protein Family
UniProt Gene Name
KRT5
UniProt Synonym Gene Names
CK-5; K5
UniProt Entry Name
K2C5_HUMAN
NCBI Description
The protein encoded by this gene is a member of the keratin gene family. The type II cytokeratins consist of basic or neutral proteins which are arranged in pairs of heterotypic keratin chains coexpressed during differentiation of simple and stratified epithelial tissues. This type II cytokeratin is specifically expressed in the basal layer of the epidermis with family member KRT14. Mutations in these genes have been associated with a complex of diseases termed epidermolysis bullosa simplex. The type II cytokeratins are clustered in a region of chromosome 12q12-q13. [provided by RefSeq, Jul 2008]
Uniprot Description
K5: Heterotetramer of two type I and two type II keratins. Keratin-5 associates with keratin-14. Interacts with TCHP. Belongs to the intermediate filament family.
Protein type: Cytoskeletal
Chromosomal Location of Human Ortholog: 12q13.13
Cellular Component: cytoplasm; cytosol; intermediate filament; keratin filament; membrane; mitochondrion; nucleus; plasma membrane
Molecular Function: protein binding; structural constituent of cytoskeleton
Biological Process: epidermis development; hemidesmosome assembly
Disease: Dowling-degos Disease 1; Epidermolysis Bullosa Simplex With Migratory Circinate Erythema; Epidermolysis Bullosa Simplex With Mottled Pigmentation; Epidermolysis Bullosa Simplex, Autosomal Recessive 1; Epidermolysis Bullosa Simplex, Dowling-meara Type; Epidermolysis Bullosa Simplex, Generalized; Epidermolysis Bullosa Simplex, Localized
Research Articles on KRT5
Similar Products
Product Notes
The KRT5 krt5 (Catalog #AAA684278) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. AAA Biotech's Cytokeratin 5/6 can be used in a range of immunoassay formats including, but not limited to, Immunohistochemistry-Paraffin (IHC-P). Dilution: 1:100 - 1:200 IHC-P Protocol - Instruction Manual: 1. Deparaffinize the section in 3 changes of xylene, 10 minutes each. 2. Wash the section in 96%, 80% and 70% ethyl alcohol for 10 minutes each. 3. Rinse in distilled water, 2 x 5 minutes. 4. Block the endogenous peroxidase by incubating the tissue in 3% hydrogen peroxide (H2O2) for 10 minutes. 5. Wash in distilled water, 2 x 5 minutes. 6. For antigen retrieval: Immerse the slide in Tris-EDTA buffer, pH 9.0 and incubate at 95-97ºC in water bath for 25 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, 2 x 5 minutes. 9. Wash in PBS (phosphate buffer saline, pH 7.0-7.5) supplemented with 0.05% of Tween-20 (buffer A), 2 x 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. 11. Wash 3 x 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). 13. Wash 3 x 5 minutes with buffer A. 14. Apply the chromogen (DAB), 1 - 3 minutes. 15. Wash in water, 2 x 5 minutes. 16. Stain in hematoxylin for 5 minutes. 17. Wash in distilled water, 3 x 2 minutes. 18. 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. Ventana Protocol - Instruction Manual: (Short Application Protocol for Ventana Benchmark Slide Staining System) 1. Deparafinization (Enter). 2. Heating glass (72°C) with the medium temperatures. Deparafinization. 3. Prolonged deparafinization (Enter). 4. Cell conditioning (Enter). 5. ULTRA Conditioner #1 (Enter). 6. Heating glass (96°C), incubation 8 min (Cell conditioner #1). 7. ULTRA CC1 solution application – 20 min (Enter). 8. Titration (Enter). 9. Hand apply - primary antibody. Incubation 32 min. 10. UltraWash (Enter). 11. Nuclear stain (Enter). 12. Hematoxylin application - one drop (nuclear stain), cover and incubate 8 min. 13. After nuclear stain (Enter). 14. Bluing reagent application, one drop. After nuclear stain, cover and incubate 4 min. Researchers should empirically determine the suitability of the KRT5 krt5 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 5/6, 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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