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mAbs Against HUMAN Protein

FOXP1

CONTACT INFORMATION:LRF Haemato-oncology Group, University of Oxford
TYPE:mouse anti human
CLONE NAME:JC12
PROTEIN:Endogenous murine Foxp1
PROTEIN WEB:http://www.ncbi.nlm.nih.gov/omim/605515
ANTIGEN USED:Not reactive with the BCLX immunogen, recognised a mouse autoantigen
FUSION PARTNER:NS1
ISOTYPE:IgG2a
SPECIES REACTIVITY:Human and mouse
PREPARATION AND STORAGE:Aliquot and store at 4oC. Do not freeze.
APP RECOMMENDED:IHQ-paraffin, IHQ-frozen, WB, IP
APP NO TESTED:Flow cytometry, IF

DESCRIPTION

The JC12 monoclonal antibody was produced by Ms Jacqueline Cordell in the LRF Immunodiagnostics Unit in Oxford. The antibody recognised a nuclear mouse autoantigen that was identified as the forkhead transcription factor FOXP1 by Dr Alison Banham. The FOXP1 gene maps to an important tumour suppressor locus on 3p14.1 and is targeted by recurrent chromosome translocations in B-cell non-Hodgkin lymphoma. FOXP1 is widely expressed in normal tissues and its expression is commonly de-regulated in malignancies. Loss of FOXP1 expression has been correlated with a poor prognosis in solid tumours, such as breast cancer. In contrast high level expression of smaller isoforms of the FOXP1 protein identifies high risk patients with diffuse large B-cell lymphoma. The JC12 antibody recognises a C-terminal epitope common to most isoforms of both the human and murine FOXP1 proteins.

PUBLICATION DESCRIBING ANTIBODY CHARACTERIZATION/VALIDATION

A.H. Banham, N. Beasley, E. Campo, P.L. Fernandez, C. Fidler, K. Gatter, M. Jones, D.Y. Mason, J.E. Prime, P.Trougouboff, K.Wood & J.L. Cordell, 2001. The FOXP1 winged helix transcription factor is a novel candidate tumour suppressor gene on chromosome 3p. Cancer Research, 61, 8820-8829.

REFERENCES

A.H. Banham, N. Beasley, E. Campo, P.L. Fernandez, C. Fidler, K. Gatter, M. Jones, D.Y. Mason, J.E. Prime, P.Trougouboff, K.Wood & J.L. Cordell, 2001. The FOXP1 winged helix transcription factor is a novel candidate tumour suppressor gene on chromosome 3p. Cancer Research, 61, 8820-8829.

C.P. Hans, D.D. Weisenburger, T.C. Greiner, R.D. Gascoyne, J. Delabie, H.K. Muller-Hermelink, E. Campo, R.M. Braziel, E.S. Jaffe, Z. Pan, P. Farinha, L.M. Smith, J.C. Lynch, B. Falini, A.H. Banham, A. Rosenwald, L.M. Staudt, J.M. Connors, J.O. Armitage, W.C. Chan, 2004. Confirmation of the molecular subclassification of diffuse large B-cell lymphoma by immunohistochemistry using a tissue microarray. Blood 103: 275-282.

W.W.L. Choi, D.D. Weisenburger, T.C. Greiner, M.A. Piris, A.H. Banham, J. Delabie, R.M. Braziel, J. Iqbal, G. Lenz, J.M. Vose, C.P. Hans, K. Fu, L.M. Smith, M. Li, Z. Liu, H. Geng, R.D. Gascoyne, A. Rosenwald, L. Rimsza, E. Campo, E.S. Jaffe, D.L. Jaye, P.A. Wade, L.M. Staudt & W.C. Chan, 2009. A new immunostain algorithm classifies diffuse large B-cell lymphoma into molecular subtypes with high accuracy. Clin Cancer Res 15: 5494-5402.

S.B. Fox, P. Brown, C. Han, S. Ashe, R.D. Leek, A.L. Harris, A.H. Banham, 2004. Expression of the forkhead transcription factor FOXP1 is associated with estrogen receptor-α and improved survival in primary human breast carcinomas. Clinical Cancer Research 10, 3521-3527.

S.L. Barrans, J. Fenton, R.G. Owen, A.H. Banham, A.S. Jack, 2004. Strong expression of FOXP1 identifies a distinct subset of diffuse large B-cell lymphoma patients with poor outcome. Blood 104, 2933-2935.

A.H. Banham, J.M. Connors, P.J. Brown, J.L. Cordell, G. Ott, G. Sreenivasan, P. Farinha, D.E. Horsman and R. Gascoyne, 2005. Expression of the FOXP1 transcription factor is strongly associated with inferior survival in patients with diffuse large B-cell lymphoma. Clin. Cancer Res. 11; 1065-1072.

P. Brown, T. Marafioti, R. Kusec, A.H. Banham, 2005. The FOXP1 transcription factor is expressed in the majority of follicular lymphomas but is rarely expressed in classical and lymphocyte predominant Hodgkin’s lymphoma. J. Molec. Histol. 36, 249-256.

A. Giatromanolaki, M.I. Koukourakis, E. Sivridis, K.C. Gatter, A.L. Harris, A.H. Banham, 2006. Loss of expression and nuclear/cytoplasmic shuttling of the FOXP1 forkhead transcription factor are common events in early endometrial cancer: relationship with estrogen receptors and HIF-1α expression. Mod. Pathol. 19, 9-16.

V.L. Sheen, R.J. Ferland, J. Neal, M. Harney, R.S.Hill, A. Banham, P. Brown, A. Chenn, J. Corbo, J. Hecht, R. Folkerth, C.A. Walsh, 2006. Neocortical neuronal arrangement in Miller Dieker syndrome. Acta Neuropathol. 111, 489-496.

P.J. Brown, R. Kagaya and A.H. Banham, 2008. Characterisation of human FOXP1 isoform 2, using monoclonal antibody 4E3-G11, and intron retention as a tissue-specific mechanism generating a novel FOXP1 isoform. Histopathol. 52: 632-7.

P.J. Brown, S.L. Ashe, E. Leich, C. Burek, S. Barrans, J. Fenton, A.S. Jack, K Pulford, A. Rosenwald and A.H Banham, 2008. Potentially oncogenic B-cell activation induced smaller isoforms of FOXP1 are highly expressed in the activated B-cell-like subtype of DLBCL. Blood 111: 2816-2824.

A. Goatly, C. Bacon, S. Nakamura, H. Ye, I. Kim, P.J. Brown, A. Ruskoné-Formestraux, P. Cervera, B. Streubel, A.H. Banham and M-Q. Du, 2008. FOXP1 abnormalities in lymphoma: translocation breakpoint mapping reveals insights into deregulated transcriptional control. Mod. Pathol. 21: 902-911.

H. Nyman, M. Jerkeman, M.-L. Karjalainen-Lindsberg, A.H. Banham & S. Leppa, 2009. Prognostic impact of activated B-cell focused classification in diffuse large B-cell lymphoma patients treated with R-CHOP. Mod. Path. 22: 1094-1101.

C. Copie-Bergman, P. Gaulard, K. Leroy, J. Briere, M. Baia, J.-P. Jais, G.A. Salles, F. Berger, C. Haioun, H. Tilly, J.-F. Emile, A.H. Banham, N. Mounier, C. Gisslebrecht, P. Feugier, B. Coffier & T.J. Molina, 2009. An Immuno-FISH Index predicts survival in diffuse large B-cell lymphoma patients treated with R-CHOP: a GELA study. J. Clin. Oncol. 27(33): 5573-5579.

M. Rayoo, M. Yan, G.J. Bates, E.A. Takano, KConFab investigators, P.J. Brown, A.H. Banham & S.B. Fox, 2009. Expression of the forkhead box transcription factor FOXP1 is associated with estrogen receptor α, estrogen receptor β and improved survival in familial breast cancers. J. Clin. Pathol. 62(10): 896-902.

P.J. Brown, A.J. Campbell, L. Lyne, J. Chi, C.H. Lawrie, R. Kusec, A.H. Banham 2009. Expression of the FOXP1 transcription factor is post-transcriptionally silenced in normal and malignant CD138+ plasma cells. The Open Leukemia Journal 3: 16-23.

E. Ballabio, J. Chi, G. Roncador, A.H. Banham, C.S.R. Hatton, C.H. Lawrie. Comparison of Choi and Hans’ algorithms by immunohistochemistry and quantitative reverse transcriptase-PCR. Clin. Cancer Res. 16(14):3805-6.

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
Recommended Result obtained is satisfactory. The reagent can be use in this application
JC12 1/10 supernatant N/A DAKO Envision Tonsil / THIEL Nucleus Mouse
Paraffin tissue
Recommended Result obtained is satisfactory. The reagent can be use in this application
JC12 1/80 supernatant Tris/EDTA DAKO Envision Tonsil / THIEL Nucleus Mouse
Immunofluorescence
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JC12 mAb in transfected cells
Nuclear and cytoplasmic staining of FOXP1 transfected COS1 cells. No reactivity with FOXP family members (FOXP2, FOXP3, FOXP4).
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JC12 IHC on paraffin tissues
JC12 staining on normal tonsil shows characteristic FOXP1 expression in mantle zone B cells, together with a proportion of germinal centre and interfollicular lymphocytes.
JC12 staining identified differential FOXP1 expression in diffuse large B-cell lymphoma, with both strongly nuclear positive cases and negative cases being identified.
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
JC12 1/30 supernatant DB / Tonsil / THIEL Approx 75kDa
Immunoprecipitation
Recommended Result obtained is satisfactory. The reagent can be use in this application
JC12 supernatant DB / Tonsil / THIEL Approx 75kDa
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JC12 Western Blotting on cell lines
JC12 Western Blotting of an activated B-cell-like diffuse large B-cell lymphoma (ABC-DLBCL) cell line (OCI-Ly3) and a panel of breast and prostate cell lines. Most cell lines express the full length FOXP1 protein, but smaller isoforms are also expressed in some. These are particularly abundant in the ABC-DLBCL cell line.