mAbs Against HUMAN Protein
GCET1
| STATUS: |  |
|---|---|
| CONTACT INFORMATION: | Monoclonal Antibodies Unit, Centro Nacional de Investigaciones Oncológicas |
| STATUS: | Validated |
| TYPE: | mouse anti human |
| CLONE NAME: | RAM |
| PROTEIN: | full length human GCET1 |
| PROTEIN WEB: | http://www.ncbi.nlm.nih.gov/protein/110225347 |
| ANTIGEN USED: | HIS-GCET1 |
| 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
GCET1 (germinal center B-cell expressed transcript 1) belongs to the SERPIN family of proteins and is located within a cluster of SERPIN genes on chromosome 14q32). This protein may play an important role in GC-B cell physiology. Its expression in lymphomas could be important in differential diagnosis and research.
Publication describing antibody characterization/validation
Montes-Moreno S, Roncador G, Maestre L, Martinez N, Sanchez-Verde L, Camacho F, Cannata J, Martinez-Torrecuadrada JL, Shen Y, Chan WC, Piris MA. Gcet1 (centerin), a highly restricted marker for a subset of Germinal Centre-derived lymphomas. Blood. 2007.http://www.ncbi.nlm.nih.gov/pubmed/17898315
References
Montes-Moreno S, Roncador G, Maestre L, Martinez N, Sanchez-Verde L, Camacho F, Cannata J, Martinez-Torrecuadrada JL, Shen Y, Chan WC, Piris MA. Gcet1 (centerin), a highly restricted marker for a subset of Germinal Centre-derived lymphomas. Blood. 2007.
Applications
| IHC Techniques | Clone | Dilution | Antibody concentration | Antigen retrieval method | Visualization kit | +/- control | Protein localization | Positivity in other species | Protocol |
|---|---|---|---|---|---|---|---|---|---|
| Frozen tissue and cytospins | |||||||||
Not recommended Reagent discarded for this application | RAM | / | |||||||
| Paraffin tissue | |||||||||
Recommended Result obtained is satisfactory. The reagent can be use in this application | RAM | 1:4 | supernatant | ER2 20 min (Tris-EDTA) | Novolink | Tonsil / | cytoplasmic | Available | |
| Immunofluorescence | |||||||||
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- Cytospin slides of transfected HEK cells.
- The transfected HEK cell line with GCET1 showed specific labelling with RAM antibody. HEK-GCET2 transfected cells were used as negative control.
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- Immunostaining of RAM antibody on human paraffin tissues.
| 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 | RAM | 1:5 | supernatant | HT cell line / MHHPREB1 cell lines | 46kDa | 46kDa | Available | |
| Immunoprecipitation | ||||||||
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- GCET1 expression by Western blot.
- The first two rows demonstate the specificity of RAM antibody on HEK transfected cells.
Lane 1 Hek-GCET1-V5 transfected cells (20ug) (+)
Lane 2 Hek-GCET2-V5 transfected cells (20ug) (-)
Lane 3 MHHPREB1 cell line (100ug) (-)
Lane 4 Karpas 422 cell line (100ug) (+)
Lane 5 DB cell line (100ug) (+)
Lane 6 HT cell line (100ug) (+)
Lane 7 OCILY3 cell line (100ug) (+)
| OTHERS | Title | Description | Protocol |
|---|---|---|---|
Recommended Result obtained is satisfactory. The reagent can be use in this application | Immunofluorescence combined with enzymatic staini |
Enlarge image
- Doble immunofluorescence combined with enzymatic staining in reactive follicles.
- Double immunostaining for KI-67 and GCET1 demonstrated that GCET1 positive cells (red) are double-positive with KI-67 (green), whereas some KI-67 positive cells are negative with GCET1.