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mAbs Against HUMAN Protein

FOXP2

Antibody Validated
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
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.
Enlarge image
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
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
Enlarge image
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.
Enlarge image
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.