mAbs Against HUMAN Protein
|CONTACT INFORMATION:||Monoclonal Antibodies Unit, Centro Nacional de Investigaciones Oncológicas|
|TYPE:||Rat anti human|
|PROTEIN:||Serine/threonine-protein kinase Pim-2|
|ANTIGEN USED:||PIM2 HIS-PIM2 recombinant protein|
|FUSION PARTNER:||myeloma p3-NS1/Ag4-1 (NS1) cells|
|PREPARATION AND STORAGE:||Aliquot and store at 4C. Do not freeze|
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].
|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
- 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.
- 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|
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|
- 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) (+)
- 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).