mAbs Against HUMAN Protein
SUZ12
| STATUS: |  |
|---|---|
| CONTACT INFORMATION: | Monoclonal Antibodies Unit, Centro Nacional de Investigaciones Oncológicas |
| STATUS: | Validated |
| TYPE: | mouse anti human |
| CLONE NAME: | 220A |
| PROTEIN: | human SUZ12 |
| PROTEIN WEB: | http://www.ncbi.nlm.nih.gov/protein/197333809 |
| ANTIGEN USED: | MBP-SUZ12 recombinant protein |
| FUSION PARTNER: | NS1/Ag4-1 (NS1) cells |
| ISOTYPE: | IgG1 |
| SPECIES REACTIVITY: | Human and mouse |
| PREPARATION AND STORAGE: | STORAGE: Aliquot and store at 4C. Do not freeze |
| APP RECOMMENDED: | IHQ-paraffin, IHQ-frozen, IF, WB |
| APP NO TESTED: | Flow cytometry, IP |
Description
This zinc finger gene has been identified at the breakpoints of a recurrent chromosomal translocation reported in endometrial stromal sarcoma. Recombination of these breakpoints results in the fusion of this gene and JAZF1. The protein encoded by this gene contains a zinc finger domain in the C terminus of the coding region.
Publication describing antibody characterization/validation
Deregulated expression of the polycomb-group protein SUZ12 target genes characterizes mantle cell lymphoma. Martín-Pérez D, Sánchez E, Maestre L, Suela J, Vargiu P, Di Lisio L, Martínez N, Alves J, Piris MA, Sánchez-Beato M. Am J Pathol 2010 Aug;177(2):930-42. http://www.ncbi.nlm.nih.gov/pubmed?term=maestre%20l%20sanchez-beato%20m
References
Deregulated expression of the polycomb-group protein SUZ12 target genes characterizes mantle cell lymphoma. Martín-Pérez D, Sánchez E, Maestre L, Suela J, Vargiu P, Di Lisio L, Martínez N, Alves J, Piris MA, Sánchez-Beato M. Am J Pathol 2010 Aug;177(2):930-42.
Applications
| IHC Techniques | Clone | Dilution | Antibody concentration | Antigen retrieval method | Visualization kit | +/- control | Protein localization | Positivity in other species | Protocol |
|---|---|---|---|---|---|---|---|---|---|
| Frozen tissue and cytospins | |||||||||
Recommended Result obtained is satisfactory. The reagent can be use in this application | 220A | neat | supernatant | no | tonsil / | nuclear | |||
| Paraffin tissue | |||||||||
Recommended Result obtained is satisfactory. The reagent can be use in this application | 220A | neat | supernatant | Tris-EDTA | tonsil / | nuclear | mouse | Available | |
| Immunofluorescence | |||||||||
Recommended Result obtained is satisfactory. The reagent can be use in this application | 220A | 1:100 | purified 1mg/ml | tonsil / | nuclear | Available | |||
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- 220A mAb in transfected cells
- Nuclear staining on frozen cytospin preparations of transfected HEK293T-V5-SUZ12 cells using antibody 220A. Anti-V5 antibody was used as positive control. HEK-V5-BTLA transfected cells were used as negative control.
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- Antibody 220A can be used to detect SUZ12 protein in human frozen tonsil.
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- 220A staining in human paraffin sections.
- SUZ12 protein is strongly expressed by proliferating cells in tonsil germinal center as well as in germinal cells of the testis.
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- Double immunofluorescence staining.
- SUZ12 (220A) nuclear protein in green and IgD membrane staining in red.
| 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 | 220A | neat | supernatant | Hela / | 83kDa | 83kDa | mouse | Available |
| Immunoprecipitation | ||||||||
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- Effect of SUZ12 RNA interference (siRNA) in Jeko-1 and Z-138 mantle cell lymphoma cell lines.
- Effect of SUZ12 RNA interference (siRNA) in Jeko-1 and Z-138 mantle cell lymphoma cell lines treated with two different hairpins against SUZ12 during three days. A control cell line (empty vector) was used as negative control. Expression of SUZ12 was analyzed using 220A mAb. A decrease of protein expression was observed in siSUZ12 Jeko-1 and Z-138 cell extracts confirming antibody specificity. Band signals were normalized with tubulin as a loading control. This image was kindly donated by Margarita Sánchez-Beato PhD.
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- 200A mAb in WB
- Lane 1 Hek-SUZ12-V5 (30цg) (+)
Lane 2 Hek-mTIMP2 (30ug) (-)
Lane 3 Hela cell line (200цg) (+)
Lane 4 Jeko1 cell line (200ug) (+)
Lane 5 human tonsil (200ug) (-)
Lane 6 human brain (200ug) (-)
Lane 7 MT-2 cell line (200ug) (+)
Lane 8 3T3 mouse cell line (200ug) (-)
Lane 9 U266 cell line (200ug) (+)
Tubulin was used as loading control.