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mAbs Against HUMAN Protein

SUZ12

Antibody Validated
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
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
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.