- Purified, prediluted
- Mouse monoclonal
- Suitable for IHC and WB
- Reacts with bovine, human, mouse and rat
- Isotype: IgG1
Product description
| Quantity | 5 ml |
|---|---|
| Antibody Type | Monoclonal |
| Host | Mouse |
| Isotype | IgG1 |
| Conjugate | Unconjugated |
| Application | IHC, WB |
| Purification | Affinity chromatography |
| Reactivity | Bovine, Human, Mouse, Rat |
| Storage | Short term at 2-8°C; long term storage in aliquots at -20°C; avoid freeze/thaw cycles |
| Intended use | Research use only |
| Clone | Ks 5+8.22/C22 |
| Immunogen | Human keratin K8, purified from SDS PAGE gel |
| Formulation | PBS pH 7.4 with 0.5% BSA and 0.09% sodium azide |
| UniprotID | Q5XQN5 (Bovine), Q7RTS7 (Human), Q922U2 (Mouse), P05786 (Bovine), P05787 (Human), P11679 (Mouse) |
| Synonym | Keratin, type II cytoskeletal 74, Cytokeratin-74, CK-74, Keratin-5c, K5C, Keratin-74, K74, Type II inner root sheath-specific keratin-K6irs4, Type-II keratin Kb37, KRT74, K6IRS4, KB37, KRT5C, KRT6IRS4, Keratin, type II cytoskeletal 8, Cytokeratin-8, CK-8, Keratin-8, K8, Type-II keratin Kb8, KRT8, CYK8 |
Applications
| Tested applications | Tested dilutions |
|---|---|
| Immunohistochemistry (IHC) - frozen | Ready-to-use |
| Immunohistochemistry (IHC) - paraffin | Ready-to-use (protease treatment and/or microwave treatment recommended) |
| Western Blot (WB) | Assay dependent |
Background
C22 represents an excellent marker for distinguishing carcinomas from all non-epithelial tumors. The antibody specifically reacts with keratins K5 and K8 present in nearly all epithelia.
Polypeptide reacting: Mr 52,500, Mr 58,000 keratins (type II keratins K5 and K8; formerly also designated cytokeratins 5 and 8) of human epithelial cells. Epitope has been mapped to aa 353-367 on alpha helical rod domain of Keratin K8 (Waseem et al., 2004).
Reactivity on cultured cell lines: MCF-7, RT 112, HT-29, HaCaT, Detroit 562, RPMI 2650, SSC-12, bovine BMGE+H, BMGE-H, MDBK.
Waseem A, Karsten U, Leigh IM, Purkis P, Waseem NH, Lane BE: Conformational changes in the rod domain of human keratin 8 following heterotypic association with keratin 18 and its implication for filament stability. Biochemistry 43, 1283-1295 (2004).
References/Publications (3)
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FAQs
The concentration of purified antibodies is mentioned on the datasheet.
For prediluted antibodies the concentration may vary from lot to lot. The concentration of these antibodies is not mentioned on the datasheet and can be requested at support@progen.com.
The supernatant format contains FCS proteins from cell culture medium supplemented with FCS.
The serum antibodies contain other proteins present in serum.
- Supernatant and supernatant concentrate: This format contains hybridoma cell culture supernatant. The antibody is not purified and the antibody concentration is not determined. The antibody concentration may vary from lot to lot. Therefore we recommend to titrate the optimal concentration for the application used for each new lot.
- Lyophilized, purified: This format contains purified antibody in lyophilized form. The reconstitution of this antibody is described in the datasheet. The buffer composition after reconstitution is also mentioned on the datasheet.
- Liquid, purified: This format contains purified antibody in liquid format. The concentration is mentioned on the datasheet.
- Prediluted, purified: This format contains purified antibody in liquid format. Most antibodies in this format are diluted to be ready-to-use for IHC with standard tissue. But some antibodies of this format need further dilution for IHC. This is mentioned on the datasheet.
Most of our liquid antibodies and reconstituted lyophilized antibodies may be stored for short term storage (up to 3 month) at 2-8°C. For long term storage we recommend to store the antibody at -20°C in aliquots. Please avoid freeze and thaw cycles.
Most of our conjugated antibodies should be stored at 2-8°C.
The individual storage conditions are mentioned on the datasheet.
Positive tissue: outer root sheath of hair follicle, sweat gland epithelium on the foot pads, filiform papillae of tongue.
- Homogenization (e.g. with polytron) of tissue samples in buffer L (140 mM NaCl, 5 mM EDTA, 5 mM EGTA, 10 mM Tris-HCl pH7.6) supplemented with protease inhibitors; use 1 ml buffer for approx. 0.1 g tissue.
- To reduce viscosity by high DNA contents, benzonase treatment can be included (30 min at 37°C).
- Centrifugation for 10 min (max. speed, table top centrifuge).
- Resuspend pellet in "Triton extraction buffer" (buffer L plus 1% Triton X-100) using e.g. a dounce homogenizer.
- Centrifugation for 10 min (max. speed, table top centrifuge).
- Resuspend pellet in "high salt extraction buffer" (buffer L plus 1.5 M KCl) using e.g. a dounce homogenizer.
- Centrifugation for 10 min (max. speed, table top centrifuge).
- Resuspend pellet in "Triton extraction buffer" (buffer L plus 1% Triton X-100) using e.g. a dounce homogenizer.
- Centrifugation for 10 min (max. speed, table top centrifuge).
- Resuspend pellet in Laemmli buffer and boil (5 min) for SDS-PAGE.
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