EuroMAbNet: European Monoclonal Antibodies NetworkEuroMAbNet. European Monoclonal Antibodies Network

☰ Menu

mAbs Against HUMAN Protein

SPIB

Antibody Validated
CONTACT INFORMATION:Monoclonal Antibodies Unit, Centro Nacional de Investigaciones Oncológicas
STATUS:Validated
TYPE:mouse anti human
CLONE NAME:235D
PROTEIN:Human SPIB
PROTEIN WEB:http://omim.org/entry/606802
ANTIGEN USED:SPIB-HIS recombinant protein (full length protein)
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
APP NO RECOMMENDED:WB, IHQ-frozen
APP NO TESTED:IF, Flow cytometry, IP

DESCRIPTION

Spi-B is a member of the Ets family of transcription factors that has high homology with the transcription factor Pu.1. The human Spi-B protein has 67% amino acid sequence identity with human Pu.1 and binds specifically to the Pu.1 binding consensus sequence 5 GAGGAA 3. Spi-B can also interact with the co-activator IRF-4 (Pip) which binds a sequence element in the Igλ enhancer when complexed with Pu.1.

PUBLICATION DESCRIBING ANTIBODY CHARACTERIZATION/VALIDATION

SPIB, a novel immunohistochemical marker for human blastic plasmacytoid dendritic cell neoplasms: characterization of its expression in major hematolymphoid neoplasms. Montes-Moreno S, Ramos-Medina R, Martínez-López A, Barrionuevo Cornejo C, Parra Cubillos A, Quintana-Truyenque S, Rodriguez Pinilla SM, Pajares R, Sanchez-Verde L, Martinez-Torrecuadrada J, Roncador G, Piris MA. Blood. 2013 Jan 24;121(4):643-7.

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
235D 1:3 supernatant 20 minutes ER2 (Tris-EDTA) Novolink kit Tonsil / nuclear Available
Immunofluorescence
Enlarge image
SPIB antibody (235D) in transfected cells.
Validation of 235D monoclonal antibody in HEK-V5-SPIB transfected cells. Anti-V5 monoclonal antibody was used as positive control. HEK-BCL7A was used as negative control.

Enlarge image
Anti-SPIB (235D) antibody on human paraffin section.
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
235D 1:2 supernatant SUDHL6 cell line / 48kDa Available
Immunoprecipitation
Enlarge image
Western Blotting of SPIB (235D) using transfected cells and different lymphoma cell lines
Lane 1 Transfected Hek-V5-SPIB (10цg) (+)
Lane 2 Transfected Hek-C-myb (10цg) (-)
Lane 3 Raji cell line (100цg) (+)
Lane 4 Oci-Ly-3 cell line (100цg) (+)
Lane 5 LP1 cell line (100цg) (-)
Lane 6 Karpas 299 cell line (100цg) (-)
Lane 7 SD-1 cell line (100цg) (-)
Lane 8 REH cell line(100цg) (-)
Lane 9 697 cell line (100цg) (-)
OTHERS
OTHERS Title Description Protocol
Recommended Result obtained is satisfactory. The reagent can be use in this application
Double SPIB/CD123 immunoenzimatic staining
Enlarge image
Double SPIB/CD123 immunoenzimatic staining
Double immunoenzymatic staining shows the expression of SPIB (brown) and CD123 (RED) in plasmacytoid dendritic cells.