mAbs Against HUMAN Protein
FOXP2
| STATUS: |  |
|---|---|
| CONTACT INFORMATION: | LRF Haemato-oncology Group, University of Oxford |
| STATUS: | Validated |
| TYPE: | mouse anti human |
| CLONE NAME: | FOXP2-73A/8 |
| PROTEIN: | N-terminal aa 1-86 of the human FOXP2 protein |
| PROTEIN WEB: | http://www.ncbi.nlm.nih.gov/omim/605317 |
| ANTIGEN USED: | Bacterially expressed GST-protein containing N-terminus of FOXP2 |
| FUSION PARTNER: | X653 |
| ISOTYPE: | IgG1 |
| SPECIES REACTIVITY: | Human and mouse |
| PREPARATION AND STORAGE: | Aliquot and store at 4oC. Do not freeze. |
| APP RECOMMENDED: | IHQ-paraffin, WB |
| APP NO RECOMMENDED: | IHQ-frozen |
| APP NO TESTED: | IP, IF, Flow cytometry |
Description
FOXP2 is a member of the forkhead transcription factor. Its mutation is associated with an inherited speech and language disorder and its biological roles have primarily been studied in neuronal tissues. FOXP2 is also aberrantly expressed in abnormal plasma cells and it may play a role in the pathogenesis of multiple myeloma. The FOXP2-73A/8 antibody detects human and murine FOXP2 and does not cross-react with other members of the FOXP family (FOXP1, FOXP3 or FOXP4).
References
A.J. Campbell, L. Lyne, P.J. Brown, R.J. Launchbury, P.A. Bignone, J. Chi, G. Roncador, C.H. Lawrie, K. Gatter, R. Kusec and A.H. Banham, 2009. Aberrant expression of the neuronal transcription factor FOXP2 in neoplastic plasma cells. Br J Haematol. 149(2): 221-30.
Applications
| IHC Techniques | Clone | Dilution | Antibody concentration | Antigen retrieval method | Visualization kit | +/- control | Protein localization | Positivity in other species | Protocol |
|---|---|---|---|---|---|---|---|---|---|
| Frozen tissue and cytospins | |||||||||
Not recommended Reagent discarded for this application | FOXP2-73A/8 | neat | supernatant | N/A | various | tonsillar epithelium / tonsillar lymphocytes | |||
| Paraffin tissue | |||||||||
Recommended Result obtained is satisfactory. The reagent can be use in this application | FOXP2-73A/8 | 1/1000 | supernatant | Tris/EDTA | Novolink | onsillar epithelium / tonsillar lymphocytes | nuclear and cytoplasmic | mouse | |
| Immunofluorescence | |||||||||
Enlarge image
- Recombinant human FOXP2 detection in transfected cells
- Immunolabelling of paraffin embedded transfected COS1 cells. The anti-FOXP2 antibody FOXP2-73A/8 recognises human FOXP2 transfectants but does not stain COS1 cells transfected with the other FOXP family members. The expression of the other family members was confirmed with independent antibodies, data shown in top right corner inset panels.
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- FOXP2 expression in reactive tonsil
- Primarily nuclear staining of paraffin embedded tonsillar epithelium is observed (left panel), although there is also some weaker cytoplasmic staining. The absence of FOXP2 expression in tonsillar plasma cells was confirmed by double labelling with VS38c shown in blue (right)
| WB Techniques | Clone | Dilution | Antibody concentration | +/- control | Expected MW | Observed Mw | Positivity in other species | Protocol |
|---|---|---|---|---|---|---|---|---|
| Western Blotting | ||||||||
Recommended Result obtained is satisfactory. The reagent can be use in this application | FOXP2-73A/8 | 1/20 | supernatant | 293T cells / JURKAT | 79.9kDa | approx 75kDa | NT | |
| Immunoprecipitation | ||||||||
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- Western blotting of FOXP transfectants
- Western blotting of COS1 cells transfected with members of the FOXP family. The FOXP2-73A/8 antibody exhibited specificity for the recombinant human FOXP2 protein. Expression of each family member in the same lysates was confirmed using other reagents (data not shown).
Enlarge image
- Silencing of endogenous FOXP2
- Silencing using FOXP2-targeted siRNA confirmed the identity of the endogenous protein in the JJN3 cell line as FOXP2.
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- Western blotting in cell lines
- Western blotting with FOXP2-73A/8 confirmed differential expression in a panel of cell lines derived primarily from haematological malignancies. Highest levels of full length protein expression were detected in cell lines derived from patients with multiple myeloma and those with Hodgkin\\\\\\\'s lymphoma.