mAbs Against HUMAN Protein
WRN
| STATUS: |  |
|---|---|
| CONTACT INFORMATION: | Monoclonal Antibodies Unit, Centro Nacional de Investigaciones Oncológicas |
| STATUS: | Validated |
| TYPE: | mouse anti human |
| CLONE NAME: | 195C |
| PROTEIN: | human WRN (RecQ protein like 2) |
| PROTEIN WEB: | http://www.hprd.org/summary?hprd_id=05212&isoform_id=05212_1&isoform_name= |
| ANTIGEN USED: | GST-WRN (aa1072-1432) |
| FUSION PARTNER: | NS1/Ag4-1 (NS1) cells |
| ISOTYPE: | IgG1 |
| SPECIES REACTIVITY: | human |
| PREPARATION AND STORAGE: | Aliquot and store at 4C. Do not freeze |
| APP RECOMMENDED: | IHQ-paraffin, IHQ-frozen, IF, WB |
| APP NO TESTED: | IP, Flow cytometry |
Description
This gene encodes a member of the RecQ subfamily and the DEAH (Asp-Glu-Ala-His) subfamily of DNA and RNA helicases. This protein contains a nuclear localization signal in the C-terminus and shows a predominant nucleolar localization. It possesses an intrinsic 3 to 5 DNA helicase activity, and is also a 3 to 5 exonuclease. Based on interactions between this protein and Ku70/80 heterodimer in DNA end processing, this protein may be involved in the repair of double strand DNA breaks. Defects in this gene are the cause of Werner syndrome, an autosomal recessive disorder characterized by premature aging.
Applications
| IHC Techniques | Clone | Dilution | Antibody concentration | Antigen retrieval method | Visualization kit | +/- control | Protein localization | Positivity in other species | Protocol |
|---|---|---|---|---|---|---|---|---|---|
| Frozen tissue and cytospins | |||||||||
| 195C | / | ||||||||
| Paraffin tissue | |||||||||
Recommended Result obtained is satisfactory. The reagent can be use in this application | 195C | 1:10 | supernatant | citrate+PK | Novolink | Testicle / | Nuclear | Available | |
| Immunofluorescence | |||||||||
Recommended Result obtained is satisfactory. The reagent can be use in this application | 195C | supernatant | / | Available | |||||
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- WRN antibody (195C) in HEK transfected cells.
- Validation of WRN195C monoclonal antibody in transfected cells. Nuclear staining on frozen cytospin preparations of transfected HEK293T-WRN cells using antibody WRN195C. Hek-CD1D transfected cells were used as negative control.
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- WRN expression in human frozen tonsil.
- In tonsil, WRN protein is expressed by ephitelial area.
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- WRN (195C) immunohistochemistry on human testicle paraffin section
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- WRN (195C) immunohistochemistry on different paraffin sections.
- Tissue samples: human tonsil, MDA-MB-231 (breast adenocarcinoma cell line), AG11395 WS (fibroblast of a patient with Werner Syndrome immortalized with SV40), Hela cell line, MRC-5 (lung fibroblast cell line) and MCF-7 (breast carcinoma cell line).
Kindly provided by Cayetano Von Kobbe.
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- WRN (195C) immunofluorescence on different cell lines.
- MDAH-2774 (Ovarian adenocarcinoma), AG-T-3141 (WS) (fibroblast WS patient) and MCF-7 (breast adenocarcinoma).
Kindly provided by Cayetano Von Kobbe.
| WB Techniques | Clone | Dilution | Antibody concentration | +/- control | Expected MW | Observed Mw | Positivity in other species | Protocol |
|---|---|---|---|---|---|---|---|---|
| Western Blotting | ||||||||
Recommended Result obtained is satisfactory. The reagent can be use in this application | 195C | supernatant | Hela / | 162kDa | 162kDa | Available | ||
| Immunoprecipitation | ||||||||
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- Western Blotting of anti-WRN (195C) using Hela WS cell lines.
- Lane 1 Hela cell line (5 ul/lane, 50,000 cells/lane) (+)
Lane 2 WS (AG11395, fibroblast of a patient with Werner Syndrome immortalized with SV40, 6 ul/lane, 60,000 cells/lane) (-)
LaminB was used as loading control.
Kindly provided by Cayetano Von Kobbe.
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- Western Blotting of anti-WRN (195C) using Hela and human heart extract.
- Lane 1 Hela cell line (200ug) (+)
Lane 2 Human heart (200ug) (-)
| OTHERS | Title | Description | Protocol |
|---|---|---|---|
Recommended Result obtained is satisfactory. The reagent can be use in this application | Epitope map of anti-WRN antibody |
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- Epitope map of anti-WRN antibody.
- The figure shows WRN protein functional domains and the corresponding sequence of the antigen used in the immunization protocol. Kindly provided by Cayetano Vonkobbe.