mAbs Against HUMAN Protein
PIM2
| STATUS: |  |
|---|---|
| CONTACT INFORMATION: | Monoclonal Antibodies Unit, Centro Nacional de Investigaciones Oncológicas |
| STATUS: | Validated |
| TYPE: | Rat anti human |
| CLONE NAME: | CHU61B |
| PROTEIN: | Serine/threonine-protein kinase Pim-2 |
| PROTEIN WEB: | http://www.ncbi.nlm.nih.gov/protein/NP_006866.2 |
| ANTIGEN USED: | PIM2 HIS-PIM2 recombinant protein |
| FUSION PARTNER: | myeloma p3-NS1/Ag4-1 (NS1) cells |
| ISOTYPE: | IgG2a |
| SPECIES REACTIVITY: | Human |
| PREPARATION AND STORAGE: | Aliquot and store at 4C. Do not freeze |
Description
This gene encodes a protooncogene that acts as a serine/threonine protein kinase. Studies determined the encoded protein functions to prevent apoptosis and to promote cell survival.[provided by RefSeq, Nov 2009].
Applications
| IHC Techniques | Clone | Dilution | Antibody concentration | Antigen retrieval method | Visualization kit | +/- control | Protein localization | Positivity in other species | Protocol |
|---|---|---|---|---|---|---|---|---|---|
| Frozen tissue and cytospins | |||||||||
| Paraffin tissue | |||||||||
Recommended Result obtained is satisfactory. The reagent can be use in this application | CHU61B | 1:20 | Tris-EDTA | Tonsil / | Available | ||||
| Immunofluorescence | |||||||||
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- CHU61B antibody in transfected cells
- Staining of CHU61B mAb in HEK293-PIM1, PIM2 and PIM3 transfected cells. No reactivity with family members PIM1 and PIM3 was found.
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- PIM2 (CHU61B) expression in human paraffin tonsil.
- CHU61B stains nuclei and cytoplasm of plasma cells and a high number of cells in the GC. In some cases it is possible to observe nuclear staining of macrophages and some endothelial cells.
| WB Techniques | Clone | Dilution | Antibody concentration | +/- control | Expected MW | Observed Mw | Positivity in other species | Protocol |
|---|---|---|---|---|---|---|---|---|
| Western Blotting | ||||||||
Inconclusive Result obtained is not completely satisfactory. More testing are needed before using these reagents in this application | CHU61B | Neat supernatat | U266 cell line / Jurkat cell line | Available | ||||
| Immunoprecipitation | ||||||||
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- Western blotting of anti-PIM2 (CHU61B) in different human cell lines.
- Lane 1 Transfected Hek-PIM1 (10ug) (-)
Lane 2 Transfected Hek-PIM2 (10ug) (+)
Lane 3 Jurkatt (100ug) (-)
Lane 4 Ramos (100ug) (+)
Lane 5 RPMI-8226 (100ug) (+)
Lane 6 OPM2 (100ug) (+)
Lane 7 U266 (100ug) (+)
Lane 8 Raji (100ug) (+)
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- Immunoprecipitation techique using CHU61B.
- Immunoprecipitation of protein extracts from HEK-PIM1, RPMI8226 and HEK-PIM2 with the anti-PIM2 commercial antibody (clone D1D2) and with anti-PIM2 CHU61B followed by western blotting with anti-PIM2 (D1D2) (1:500) and the CHU61B (neat). In both cases it is possible to observe a specific band of 34 kDa in HEK-PIM2 cell extract confirming antibody specificity. In case of RPMI8226 cell line immunoprecipitated using anti-PIM2 (clone D1D2) it is possible to observe the presence of three bands corresponding to the three PIM2 isoforms (34, 38, and 40 kDa).