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mAbs Against HUMAN Protein

PIM2

Antibody Validated
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
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
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) (+)
Enlarge image
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).