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Key Specifications Table
| Species Reactivity | Key Applications | Host | Format | Antibody Type |
|---|---|---|---|---|
| H | ICC, IHC | M | Purified | Monoclonal Antibody |
| Description | |
|---|---|
| Catalogue Number | MAB3404 |
| Replaces | 04-586 |
| Brand Family | Chemicon® |
| Trade Name |
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| Description | Anti-Cytokeratin 18 Antibody, clone CK2 |
| Product Information | |
|---|---|
| Format | Purified |
| Presentation | Liquid in 0.02 M Phosphate buffer, 0.25 M NaCl with 0.1% sodium azide. |
| Quality Level | MQ100 |
| Applications | |
|---|---|
| Application | This Anti-Cytokeratin 18 Antibody, clone CK2 is validated for use in IC, IH for the detection of Cytokeratin 18. |
| Key Applications |
|
| Application Notes | Immunocytochemistry: 20 μg/mL Immunohistochemistry: 20 μg/mL Not for formaldehyde fixed tissues. Optimal working dilutions must be determined by end user. Immunohistochemistry/Immunocytochemistry Protocols: Ideal specimens are obtained from frozen sections from shockfrozen tissue samples. The frozen sections are dried in the air and then fixed with acetone at -15 to -25°C for 10 min. Excess acetone is allowed to evaporate at 15-25°C. Material fixed in alcohol and embedded in paraffin can, however, be used, see (4). For the immunohistochemical detection of cytokeratin No. 18 in tissue sections, the tissue should not be fixed in formaldehyde as this fixative markedly reduces the staining intensity of the cytokeratin filaments. lt is advantageous to block unspecific binding sites by overlaying the sections with fetal calf serum for 20-30 min at 15-25°C. Excess of fetal calf serum is removed by decanting before application of the antibody solution. Cytocentrifuge preparations of single cells or cell smears are also fixed in acetone. These preparations should, however, not be dried in the air. Instead, the excess acetone is removed by briefly washing in phosphate-buffered saline (PBS). Further treatment is then as follows: • Overlay the preparation with 10-20 μL antibody solution and incubate in a humid chamber at 37°C for 1h. • Dip the slide briefly in PBS and then wash 3 times in PBS for 3 min (using a fresh PBS bath in each case). • Wipe the margins of the preparation dry and overlay the preparation with 10-20 μL of an anti-mouse-IgG-FITC or anti-mouse-IgG-peroxidase antibody and allow to incubate for 1 h at 37°C in a humid chamber. • Wash the slide as described above. The preparation must not be allowed to dry out during any of the steps. lf using an indirect immunofluorescence technique, the preparation should be overlaid with a suitable embed-ding medium (e. g. Moviol, Hoechst) and examined under the fluorescence microscope. lf a POD-conjugate has been used as the secondary antibody, the preparation should be overlaid with a substrate solution (see below). and incubated at 15-25°C until a clearly visible redbrown color develops. A negative control (e. g. only the secondary antibody) should remain unchanged in color during this incubation period. Wash away the substrate with PBS and stain the preparation, if desired, with hemalum stain, for about 1 min. The hemalum solution is washed off with PBS, the peparation is embedded and examined. Substrate solutions: Aminoethyl-carbazole Dissolve 2mg 3-amino-9-ethylcarbazole with 1.2 mL dimethylsulfoxide and add 28.8 mL Tris-HCI, 0.05 M; pH 7.3, and 20 μL 3% H 2 O 2 (w/v). Prepare solution freshly each day. Diaminobenzidine Dissolve 25 mg 3,3-diaminobenzidine with 50 ml Tris-HCI, 0.05 M; pH 7.3, and add 40 μL H 2 O 2 , 3% (w/v). Prepare solution freshly each day. |
| Biological Information | |
|---|---|
| Immunogen | Cytoskeletal preparation of HeLa cells (Debus et al., 1982). |
| Clone | CK2 |
| Concentration | Please refer to the Certificate of Analysis for the lot-specific concentration. |
| Host | Mouse |
| Specificity | The antibody reacts with cytokeratin No. 18 from human. This antibody is used for staining research samples of a wide variety of simple epithelia and simple glandular epithelia (e. g. intestinal, respiratory and urinary epithelia) and transitional epithelia (e. g. bladder), but stratified squamous epithelia are not stained (e. g. esophagus, epidermis). Keratin filaments of cell lines such as HeLa cells are stained (Debus et al., 1982). For staining research samples of human tumor material with this antibody, see (Debus et al., 1984). For the cytokeratin pattern of different tissues, see (Moll et al., 1982). |
| Isotype | IgG1 |
| Species Reactivity |
|
| Antibody Type | Monoclonal Antibody |
| Entrez Gene Number |
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| Entrez Gene Summary | KRT18 encodes the type I intermediate filament chain keratin 18. Keratin 18, together with its filament partner keratin 8, are perhaps the most commonly found members of the intermediate filament gene family. They are expressed in single layer epithelial tissues of the body. Mutations in this gene have been linked to cryptogenic cirrhosis. Two transcript variants encoding the same protein have been found for this gene. |
| Gene Symbol |
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| UniProt Number |
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| UniProt Summary | FUNCTION: SwissProt: P05783 # Involved in the uptake of thrombin-antithrombin complexes by hepatic cells (By similarity). When phosphorylated, plays a role in filament reorganization. Involved in the delivery of mutated CFTR to the plasma membrane. Together with KRT8, is involved in interleukin-6 (IL-6)-mediated barrier protection. SIZE: 430 amino acids; 48058 Da SUBUNIT: Heterotetramer of two type I and two type II keratins. Keratin-18 associates with keratin-8. Interacts with the thrombin- antithrombin complex (By similarity). Interacts with PNN, HCV core protein and mutated CFTR. Interacts with YWHAE, YWHAH and YWHAZ only when phosphorylated. Interacts with DNAJB6, TCHP and TRADD. SUBCELLULAR LOCATION: Cytoplasm, perinuclear region. TISSUE SPECIFICITY: Expressed in colon, placenta, liver and very weakly in exocervix. Increased expression observed in lymph nodes of breast carcinoma. PTM: Phosphorylation at Ser-34 increases during mitosis. Hyperphosphorylated at Ser-53 in diseased cirrhosis liver. Phosphorylation increases by IL-6. & Proteolytically cleaved by caspases during epithelial cell apoptosis. Cleavage occurs at Asp-238 by either caspase-3, caspase-6 or caspase-7.DISEASE:SwissProt: P05783 # Defects in KRT18 are a cause of cryptogenic cirrhosis [MIM:215600]. SIMILARITY: SwissProt: P05783 ## Belongs to the intermediate filament family. MISCELLANEOUS: There are two types of cytoskeletal and microfibrillar keratin: I (acidic; 40-55 kDa) and II (neutral to basic; 56-70 kDa). |
| Product Usage Statements | |
|---|---|
| Usage Statement |
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| Storage and Shipping Information | |
|---|---|
| Storage Conditions | The lyophilized antibody is stable when stored at -20°C.Store the reconstituted antibody solution at 2-8°C, for up to 12 months.DO NOT FREEZE. |
| Packaging Information | |
|---|---|
| Material Size | 40 µg |