mAbs Against HUMAN Protein
XBP1s
| STATUS: |  |
|---|---|
| CONTACT INFORMATION: | Monoclonal Antibodies Unit, Centro Nacional de Investigaciones Oncológicas |
| STATUS: | Validated |
| TYPE: | mouse anti human |
| CLONE NAME: | 143F |
| PROTEIN: | human XBP1s |
| PROTEIN WEB: | http://www.ncbi.nlm.nih.gov/protein/118640873 |
| ANTIGEN USED: | XBP1s C-terminus (amino-acids 165-367) |
| FUSION PARTNER: | NS1/Ag4-1 (NS1) cells |
| ISOTYPE: | IgG2a |
| SPECIES REACTIVITY: | human |
| PREPARATION AND STORAGE: | Aliquot and store at 4C. Do not freeze |
| APP RECOMMENDED: | IHQ-paraffin, IF, WB |
| APP NO RECOMMENDED: | IHQ-frozen |
| APP NO TESTED: | IP, Flow cytometry |
Description
Xbox binding protein-1 (XBP-1) is a transcription factor essential for plasma cell differentiation. XBP-1 is subject to alternative RNA processing, generating two mRNA transcripts encoding the same N-terminal DNA binding domain, but different C-terminal transactivation domains. The shorter spliced transcript, designated XBP-1s, possesses enhanced transactivation potential and stability relative to the product of the unspliced transcript, designated XBP-1u. Recent studies have uncovered several functions for XBP-1 and have implicated XBP-1 overexpression in human carcinogenesis and tumour growth under hypoxic conditions. Specifically, elevated XBP-1 mRNA levels have been detected in hepatocellular carcinomas and in primary ERα-positive breast tumors.
Publication describing antibody characterization/validation
Maestre L, Tooze R, Cañamero M, Montes-Moreno S, Ramos R, Doody G, Boll M, Barrans S, Baena S, Piris MA, Roncador G.
XBP1(S) provides a specific marker of stress response and plasma cell differentiation. Haematologica. 2009 Mar;94(3):419-22. Epub
2009 Jan 27. http://www.ncbi.nlm.nih.gov/pubmed/19176362
References
Maestre L, Tooze R, Cañamero M, Montes-Moreno S, Ramos R, Doody G, Boll M, Barrans S, Baena S, Piris MA, Roncador G.
XBP1(S) provides a specific marker of stress response and plasma cell differentiation. Haematologica. 2009 Mar;94(3):419-22. Epub
2009 Jan 27.
Montes-Moreno S, Gonzalez-Medina AR, Rodriguez-Pinilla SM, Maestre L, Sanchez-Verde L, Roncador G, Mollejo M, García JF,
Menarguez J, Montalbán C, Ruiz-Marcellan MC, Conde E, Piris MA. Haematologica. Aggressive large B-cell lymphoma with plasma
cell differentiation: immunohistochemical characterization of plasmablastic lymphoma and diffuse large B-cell lymphoma with partial
plasmablastic phenotype. 2010 Aug; 95(8):1342-9.
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 | 143F | 1:100 | supernatant | 20 minutes ER2 (Tris-EDTA) | Novolink kit | Tonsil / | Available | ||
| Immunofluorescence | |||||||||
Recommended Result obtained is satisfactory. The reagent can be use in this application | 143F | supernatant | / | Available | |||||
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- XBP1s antibody (143F) in transfected cells
- Validation of 143F monoclonal antibody in Hek-GFP-XBP1s transfected cells. Another HEK transfected cells were used as negative control.
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- XBP1s (143F) immunohistochemistry on human paraffin sections.
- In tonsil, XBP1s protein is strongly expressed by plasma cells present in germinal center B cells and in subepithelial areas. XBP1s is also expressed in plasma cells of diferent normal tissues as bone marrow. XBP1s immunostaining of lymphoid neoplasm are represented in the images below.
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- Expression of XBP1s (143F) antigen by lymphoid cells.
- Double immunofluorescent staining shows that many of the XBP1s positive cells (green) in the germinal center cells do not co-express CD20 antigen (red).
| WB Techniques | Clone | Dilution | Antibody concentration | +/- control | Expected MW | Observed Mw | Positivity in other species | Protocol |
|---|---|---|---|---|---|---|---|---|
| Western Blotting | ||||||||
| 143F | 1:10 | supernatant | SK-MM-2 cell line / Ocily3 cell line | 54kDa | Available | |||
| Immunoprecipitation | ||||||||
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- Western blotting of XBP1s (clone143F) antibody.
- Specificity of the XBP1s antibody is demonstrated by detection of a single 54 kDa band in transfectants.
Lane 1 Hek-XBP1s trasfected cells (20ug) (+)
Lane 2 Human testicle (100ug) (-)
Lane 3 GST-XBP1s recombinant protein (0.1ug) (+)
Lane 4 Karpas 620 cell line (100ug) (+)
| OTHERS | Title | Description | Protocol |
|---|---|---|---|
Recommended Result obtained is satisfactory. The reagent can be use in this application | Immunoenzimatic staining |
Enlarge image
- Double XBP1s/CD138 immunoenzimatic staining
- Double immunoenzymatic staining shows the expression of XBP1s (brown) and CD138 (red). This reveals that the mayority of XBP1s positive cells co-express CD138.