Anti-Cytokeratin 5, 6, clone D5/16B4
| Species Reactivity | Key Applications | Host | Format | Antibody Type |
|---|---|---|---|---|
| H | WB, IH(P) | Mouse | Purified | Monoclonal Antibody |
UniProt Number:
UniProt Summary
SIZE: 590 amino acids; 62378 Da
SUBUNIT: Heterotetramer of two type I and two type II keratins. Keratin-5 associates with keratin-14. Interacts with TCHP.
DISEASE: SwissProt: P13647 # Defects in KRT5 are a cause of epidermolysis bullosa simplex (EBS) [MIM:131760, 131800, 131900, 131960]. Epidermolysis bullosa (EB) is a mendelian disorder that can be inherited in an autosomal dominant or recessive form and is characterized by blistering or erosion of the skin and mucou ... see more »
SUBUNIT: Heterotetramer of two type I and two type II keratins. Keratin-5 associates with keratin-14. Interacts with TCHP.
DISEASE: SwissProt: P13647 # Defects in KRT5 are a cause of epidermolysis bullosa simplex (EBS) [MIM:131760, 131800, 131900, 131960]. Epidermolysis bullosa (EB) is a mendelian disorder that can be inherited in an autosomal dominant or recessive form and is characterized by blistering or erosion of the skin and mucou ... see more »
Entrez Gene Number:
Entrez Gene Summary
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
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Description:
Anti-Cytokeratin 5, 6, clone D5/16B4
Background Information:
The cytokeratins are a family of intermediate filaments found in almost all epithelial cells. They are divided into four subclasses depending on their patterns of reactivity and electrophoretic characteristics and are also arranged I a numbered series of 20 or more acoring to molecular weight and isoelectric point (Quinlan et al. 1985). Types 5 and 6 of cytokeratin (CK5/6) are in high frequency in mesothelial cells, as opposed to cells of adenocarcinoma. CK5/6 has been shown to be a reliable marker for mesothelioma and squamous cell carcinoma of the lung and Bowen's disease. It does not react with pulmonary adenocarcinoma.
Clone:
D5/16B4
Specificity:
Reacts with human cytokeratin 5 on immunoblots of cytoskeletal preparations of epidermis and non-cytokeratinizing epithelium (Lobeck et al., 1989) and on tissue sections. Shows reactivity with cytokeratin 6 and weak reactivity with cytokeratin 4 on immunoblots. No cross-reactivity with cytokeratins 1, 7, 8, 10, 13, 14, 18 or 19. MAB1620 recognizes basal cells and a part of stratum spinosum in the normal pavement epithelium. The antibody may be useful for distinction of low differentiated pavement epithelium carcinoma and adenocarcinoma. The antibody works on formalin fixed tissues.
Key Applications:
- Western Blotting
- Immunohistochemistry (Paraffin)
Application Notes:
Immunohistochemistry (frozen or paraffin sections): for paraffin, high-temperature antigen retrieval required (Pressure cooker, Citrate/EDTA buffer pH 6.0, 4 minutes at full pressure) 1:50-1:100 dilution.
Western blot: Recognizes a 58kDa and 56kDa band corresponding to cytokeratin 5 and 6.
Immunofluorescence
Optimal working dilutions must be determined by end user.
APPLICATION NOTES FOR MAB1620
Paraffin Embedded Sections: Formalin-fixed paraffin embedded sections are dewaxed. Before adding the antibody the section should be treated with pronase at 37°C.
Frozen Sections: Ideal frozen sections (4-5 mm) are obtained from shock frozen tissue samples. The frozen sections are air-dried and then fixed with acetone for 10 minutes at -20°C. Excess acetone is allowed to evaporate.
1) If necessary, block unspecific binding sites by overlaying the sections with 20 mL fetal calf serum for 30 minutes at 37°C in a humid chamber.
2) Cover the preparation according to the size of the section with a suitable volume of properly diluted MAB1620 and incubate for one hour at room temperature in a humid chamber.
3) Immerse the slides briefly in PBS and wash three times for three minutes each in PBS.
4) Wipe slides dry except area of section. Cover the section with 10-20 mL of a solution of anti-Mouse IgG-FITC or anti-Mouse IgG-enzyme and incubate for one hour at room temperature or at 37°C in a humid chamber .
5) Wash the slide three times for three minutes each in PBS.
The section should not be allowed to dry out during the steps. If using an enzyme conjugate, the preparation is covered with a substrate solution and incubated at room temperature until a clearly visible color develops.
Western blot: Recognizes a 58kDa and 56kDa band corresponding to cytokeratin 5 and 6.
Immunofluorescence
Optimal working dilutions must be determined by end user.
APPLICATION NOTES FOR MAB1620
Paraffin Embedded Sections: Formalin-fixed paraffin embedded sections are dewaxed. Before adding the antibody the section should be treated with pronase at 37°C.
Frozen Sections: Ideal frozen sections (4-5 mm) are obtained from shock frozen tissue samples. The frozen sections are air-dried and then fixed with acetone for 10 minutes at -20°C. Excess acetone is allowed to evaporate.
1) If necessary, block unspecific binding sites by overlaying the sections with 20 mL fetal calf serum for 30 minutes at 37°C in a humid chamber.
2) Cover the preparation according to the size of the section with a suitable volume of properly diluted MAB1620 and incubate for one hour at room temperature in a humid chamber.
3) Immerse the slides briefly in PBS and wash three times for three minutes each in PBS.
4) Wipe slides dry except area of section. Cover the section with 10-20 mL of a solution of anti-Mouse IgG-FITC or anti-Mouse IgG-enzyme and incubate for one hour at room temperature or at 37°C in a humid chamber .
5) Wash the slide three times for three minutes each in PBS.
The section should not be allowed to dry out during the steps. If using an enzyme conjugate, the preparation is covered with a substrate solution and incubated at room temperature until a clearly visible color develops.
Species Reactivity:
Human
Isotype:
IgG1
Immunogen:
Purified cytokeratin 5 (Wild & Mischke, 1986; Mischke & Wild, 1987)
Molecular Weight:
Cytokeratin 4 (59 kDa), Cytokeratin 5 (58 kDa) and Cytokeratin 6 (56 kDa)
Control:
A431 cell lysate, human bladder tumor tissue
Presentation:
Purified immunoglobulin. Liquid in 0.02M Phosphate buffer, 0.25M NaCl, pH 7.6 with 0.1% sodium azide.
Storage Conditions:
Maintain material at 2–8°C in undiluted aliquots for up to 6 months from date of receipt.
Gene Symbol:
- KRT5
- K5
- EBS2
- DDD
- Keratin-5
- KRT5A
- Cytokeratin-5
- CK5
- CK-5
Antibody Category:
Cell Structure
Antibody Sub-Category:
Cytokeratins
Usage Statement:
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Concentration:
1 mg/mL
Purification Method:
Protein A Purified
Format:
Purified
Format Code:
Pur
Antibody Type:
Monoclonal Antibody
Protein:
Cytokeratin: Cytokeratin 5
Host:
Mouse