- Purified, lyophilized
- Mouse monoclonal
- Suitable for ELISA, IHC and WB
- Reacts with human, mouse and pig
- Isotype: IgG1
| Quantity | 50 µg |
|---|---|
| Antibody Type | Monoclonal |
| Host | Mouse |
| Isotype | IgG1 |
| Conjugate | Unconjugated |
| Application | ELISA, IHC, WB |
| Purification | Affinity chromatography |
| Reactivity | Human, Mouse, Pig |
| Storage before reconstitution | 2-8°C until indicated expiry date |
| Storage after reconstitution | Up to 3 months at 2-8°C; long term storage in aliquots at -20°C; avoid freeze/thaw cycles |
| Intended use | Research use only |
| Clone | IT-Ks20.10 |
| Immunogen | Electrophoretically purified keratin K20 from human intestinal mucosa |
| Concentration | 50 µg/ml after reconstitution with 1 ml dist. water |
| Formulation | Lyophilized; reconstitute in 1 ml dist. water (final solution contains 0.09% sodium azide, 0.5% BSA in PBS buffer, pH 7.4) |
| Synonym | Cytokeratin 20 |
| Tested applications | Tested dilutions |
|---|---|
| Immunohistochemistry (IHC) - frozen | 1:50-1:100 |
| Immunohistochemistry (IHC) - paraffin | 1:50-1:100 (microwave treatment recommended) |
| ELISA | Assay dependent |
| Western Blot (WB) | Assay dependent |
IT-Ks20.10 represents an excellent marker for certain types of carcinomas such as adenocarcinomas of the colon, transitional cell carcinomas of the bladder and Merkel cell tumors of the skin. Very sensitive detection of intestestinal and gastric foveolar epithelium, urothelial umbrella cells, Merkel cells of epidermis as well as tumors originating therefrom (e.g. primary and metastatic colorectal carcinoma). Adenocarcinomas of breast, lung, endometrium and ovary (non-mucinous) as well as neuroendocrine tumors of the lung are essentially negative.
Polypeptide recognized: protein IT (keratin K20; Mr 46,000; formerly also designated cytokeratin 20).
Reactivity on cultured cell lines: HT-29, LoVo, DLD-1, SW 1116, CaCo-2, RT-4.
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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