mAbs Against HUMAN Protein
SPIB
| STATUS: |  |
|---|---|
| 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 | 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 | |||||||||
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- 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.
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- Anti-SPIB (235D) antibody on human paraffin section.
| 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 | ||||||||
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- 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 | Title | Description | Protocol |
|---|---|---|---|
Recommended Result obtained is satisfactory. The reagent can be use in this application | Double SPIB/CD123 immunoenzimatic staining |
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- Double SPIB/CD123 immunoenzimatic staining
- Double immunoenzymatic staining shows the expression of SPIB (brown) and CD123 (RED) in plasmacytoid dendritic cells.