Mouse monoclonal antibody against POKEMON: P001
Mouse monoclonal antibody against POKEMON (also known as Zinc Finger and BTB domain-containing protein 7A (ZBTB7A), Leukemia/lymphoma-related factor (LRF), POZ and Krueppel erythroid myeloid ontogenic factor and POK erythroid myeloid ontogenic factor and FBI-1).
Product Number: P001
Concentration: 0.50 mg/ml.
Storage Buffer: Phosphate buffered saline, pH 7.2 with 0.05% sodium azide.
Immunogen: 14 amino acid peptide whose sequence corresponds to a region that is specific to POKEMON (ZBTB7A) and has 100% homology with POKEMON from several species including mouse, rat, guinea pig, bats, dog, sheep, cow, primates and human.
Isotype: Mouse IgG1, Kappa
Reactivity: The antibody was tested against several human cancer cell lines, formalin fixed paraffin embedded (FFPE) human tumor xenografts & a limited number of normal human tissues. Not tested in other species.
Applications: Indirect immunofluorescence, immunohistochemistry & western blotting. Not tested for use in immunoprecipitation. Suggested starting concentrations are as follows; Western blotting 1 -2ug/ml, immunohistochemistry 2-5ug/ml. The BC17-2.9F antibody may be used on tissue culture cells grown on chamber slides, cytospins, cryosections and formalin fixed paraffin embedded (FFPE) sections. For staining of FFPE sections, use of an antigen retrieval procedure following de-paraffinization and rehydration of the tissue sections is recommended. Cultured cells and cryosections should be fixed in 1-2% paraformaldehyde for 30 mins, permeabilized in 0.25% Triton X 100 in PBS for 30 mins and non-specific binding blocked with 1% BSA in PBS. The primary antibody may be used at 3-5ug/ml in 1%BSA in PBS.
Preparation and Storage: The monoclonal antibody was purified from tissue culture supernatant using Protein G sepharose columns. Store undiluted at 4° C or aliquot and store at -20C for long term storage. Repeated freeze thaw cycles should be avoided.
A B C
- Weak nuclear, perinuclear and cytoplasmic staining of MDA-MB-231 human breast carcinoma cells is shown. The cells were stained with 5ug/ml BC17-2.9F antibody and FITC conjugated anti- mouse Ig secondary antibody. While Pokemon has a nuclear localization signal, intense staining of perinuclear and cytoplasmic areas are observed. Whether this distribution reflects pathology remains to be investigated.
- Nuclear staining of 1643 neuroblastoma cells is shown in formalin fixed paraffin embedded tumor xenograft grown in mice. The sections were stained with 5ug/ml BC17-2.9F antibody and peroxidase conjugated anti-mouse Ig secondary antibody with DAB substrate. The background staining reflects anti-mouse secondary antibody also reacting with endogenous mouse serum immunoglobulins present in the mouse grown xenografts tissue.
- Nuclear and cytoplasmic staining of reactive tonsil (human) is shown on formalin fixed paraffin tissue. The sections were stained with 5ug/ml BC17-2.9F antibody, peroxidase conjugated anti-mouse Ig secondary antibody with DAB substrate.
Western blotting- Use the BC17-2.9F antibody at 2-3 ug/ml in Tris buffered Saline with 0.05% Tween 20 and 5% non-fat dry milk or similar diluents.
Predicted Molecular size: 62 kDa.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Caution: The product contains 0.05% sodium azide. Sodium azide produces highly toxic hydrazoic acid under acidic conditions. It is therefore recommended to dilute azide compounds in running water before discarding to prevent the accumulation of potentially explosive deposits in the plumbing.
Suggested positive control cell lines: Various cancer cell lines may be used including MCF7 (ATCC HTB-22) and MDA-MB 231 (ATCC HTB-26) human breast cancer lines, HeLa human cervical carcinoma (ATCC CCL-2), or HepG2 human hepatocellular carcinoma (ATCC HB-8065). The antibody reacts with a band of approximate molecular size 62kD.
Pokemon is a 584 a.a. protein with zinc finger and BTB-POZ domains. It is a member of the POK family of proteins that play important roles in embryonic development, cell lineage determination and differentiation, but are also involved in oncogenesis. Thus Pokemon is amplified or over-expressed in numerous types of cancers, including breast and prostate cancer, lymphoma, small cell lung carcinoma and malignant glioma. It functions as a transcriptional repressor of the p19 arf p53, and RB tumor suppressor genes and works in concert with oncogenes and other proteins associated with the promotion of cell growth. Its overexpression has been shown to cause activation of membrane type 1-matrix metalloproteinase which is associated with adverse outcome in ovarian cancers and increased drug resistance. These properties are thought to impart its role in oncogenesis. In this context, Pokemon is regarded as a marker of aggressive tumors and may thus serve as a useful marker of poor prognosis in cancers. Pokemon is also being reviewed as a target for inhibiting oncogenesis.
Davis JM, Hawe N, Kabarowski J, Huang Q-H, Zhu J, Brand NJ, Leprince D, Dhordain P, Cook M, Morriss-Kay G, Zelent A. Novel BTB/POZ domain zinc-finger protein, LRF, is a potential target of the LAZ-3/BCL-6 oncogene. Oncogene .1999; 18:365-375.
Morrison DJ, Pendergrast PS, Stavropoulos P, Colmenares SU, Kobayashi R, Hernandez N. FBI-1, a factor that binds to the HIV-1 inducer of short transcripts(IST), is a POZ domain protein. Nucleic Acids Res. 1999; 27(5):1251-62.
Lee DK, Suh D, Edenberg HJ, Hur MW. POZ domain transcription factor, FBI-1, represses transcription of ADH5/FDH by interacting with the zinc finger and interfering with DNA binding activity of Sp1. J Biol Chem. 2002;277(30):26761-8.
Pendergrast PS, Wang C, Hernandez N, Huang S. FBI-1 can stimulate HIV-1 Tat activity and is targeted to a novel subnuclear domain that includes the Tat-P-TEFb-containing nuclear speckles. Mol Biol Cell. 2002; 13(3):915-29.
Pessler F, Hernandez N. Flexible DNA binding of the BTB/POZ-domain protein FBI-1. J Biol Chem. 2003; 278(31):29327-35.
Laudes M, Christodoulides C, Sewter C, Rochford JJ, Considine RV, Sethi JK, Vidal-Puig A, O'Rahilly S. Role of the POZ zinc finger transcription factor FBI-1 in human and murine adipogenesis. J Biol Chem. 2004; 279(12):11711-8.
Lee DK, Kang JE, Park HJ, Kim MH, Yim TH, Kim JM, Heo MK, Kim KY, Kwon HJ, Hur MW. FBI-1 enhances transcription of the nuclear factor-kappaB (NF-kappaB)-responsive E-selectin gene by nuclear localization of the p65 subunit of NF-kappaB. J Biol Chem. 2005; 280(30):27783-91.
Maeda T, Hobbs RM, Pandolfi PP. The transcription factor Pokemon: A new key player in cancer pathogenesis. Cancer Research: 2005; 65. 8575-8578.
Maeda T, Hobbs RM, Merghoub, Guernah I, Zelent A, Cardon-Cardo C, Teruya-Feldstein J, Pandolfi PP. Role of the proto-oncogene Pokemon in cellular transformation and ARF repression. Nature.2005; 433:278-285.
Agrawal A, Yang J, Murphy RF, Agrawal DK. Regulation of the p14ARF-Mdm2-p53 pathway: an overview in breast cancer. Exp Mol Pathol. 2006 Oct;81(2):115-22. Epub 2006. Review.
Apostolopoulou K, Pateras IS, Evangelou K, Tsantoulis PK, Liontos M, Kittas C, Tiniakos DG, Kotsinas A, Cordon-Cardo C, Gorgoulis VG. Gene amplification is a relatively frequent event leading to ZBTB7A (Pokemon) overexpression in non-small cell lung cancer. J Pathol. 2007;213(3):294-302.
Stogios PJ, Chen L, Privé GG. Crystal structure of the BTB domain from the LRF/ZBTB7 transcriptional regulator. Protein Sci. 2007; 16(2):336-42.
Zhao Z, Wang S, Zhang T. Zhongguo Fei Ai Za Zhi. [Expression and clinical significance of Pokemon in non-small cell lung cancer]. 2007;10(6):491-4.
Jeon BN, Yoo JY, Choi WI, Lee CE, Yoon HG, Hur MW. Proto-oncogene FBI-1 (Pokemon/ZBTB7A) represses transcription of the tumor suppressor Rb gene via binding competition with Sp1 and recruitment of co-repressors. J Biol Chem. 2008; 283(48):33199-210.
Laudes M, Bilkovski R, Oberhauser F, Droste A, Gomolka M, Leeser U, Udelhoven M, Krone W. Transcription factor FBI-1 acts as a dual regulator in adipogenesis by coordinated regulation of cyclin-A and E2F-4. J Mol Med (Berl). 2008; 86(5):597-608.
Salas A, Vega A, Milne RL, García-Magariños M, Ruibal A, Benítez J, Carracedo A. The 'Pokemon' (ZBTB7) Gene: No Evidence of Association with Sporadic Breast Cancer. Clin Med Oncol. 2008;2:357-62.
Zhao ZH, Wang SF, Yu L, Wang J, Chang H, Yan WL, Fu K, Zhang J. Expression of transcription factor Pokemon in non-small cell lung cancer and its clinical significance. Chin Med J (Engl). 2008;121(5):445-9.
Zhao ZH, Wang SF, Yu L, Wang J, Chang H, Yan WL, Zhang J, Fu K. Overexpression of Pokemon in non-small cell lung cancer and foreshowing tumor biological behavior as well as clinical results. Lung Cancer. 2008;62(1):113-9.
Zhao ZH, Wang SF, Yu L, Wang J, Cong DG, Chang H, Wang XF, Zhang TW, Zhang J, Fu K, Jiang JY. Zhonghua Yi Xue Za Zhi. [Effect of cisplatin on the expression of Pokemon gene: experiment with different human lung cancer cells]. 2008;88(17):1211-3. Chinese.
Choi WI, Jeon BN, Yun CO, Kim PH, Kim SE, Choi KY, Kim SH, Hur MW. Proto-oncogene FBI-1 represses transcription of p21CIP1 by inhibition of transcription activation by p53 and Sp1. J Biol Chem. 2009; 284(19):12633-44.
Choi WI, Kim Y, Kim Y, Yu MY, Park J, Lee CE, Jeon BN, Koh DI, Hur MW. Eukaryotic translation initiator protein 1A isoform, CCS-3, enhances the transcriptional repression of p21CIP1 by proto-oncogene FBI-1 (Pokemon/ZBTB7A). Cell Physiol Biochem. 2009; 23(4-6):359-70.
Aggarwal A, Hunter WJ 3rd, Aggarwal H, Silva ED, Davey MS, Murphy RF, Agrawal DK. Expression of leukemia/lymphoma-related factor (LRF/POKEMON) in human breast carcinoma and other cancers. Exp Mol Pathol. 2010; 89(2):140-8.
He S, Liu F, Xie Z, Zu X, Xu W, Jiang Y. P-Glycoprotein/MDR1 Regulates Pokemon Gene Transcription Through p53 Expression in Human Breast Cancer Cells. Int J Mol Sci. 2010; 11(9):3309-051.
Jiang L, Siu MK, Wong OG, Tam KF, Lam EW, Ngan HY, Le XF, Wong ES, Chan HY, Cheung AN. Overexpression of proto-oncogene FBI-1 activates membrane type 1-matrix metalloproteinase in association with adverse outcome in ovarian cancers. Mol Cancer. 2010; 9:318.
Tian Z, Wang H, Jia Z, Shi J, Tang J, Mao L, Liu H, Deng Y, He Y, Ruan Z, Li J, Wu Y, Ni B. Tumor-targeted inhibition by a novel strategy - mimoretrovirus expressing siRNA targeting the Pokemon gene. Curr Cancer Drug Targets. 2010; 10(8):932-41.
Aggarwal H, Aggarwal A, Agrawal DK. Epidermal growth factor increases LRF/Pokemon expression in human prostate cancer cells. Exp Mol Pathol. 2011; 91(2):496-501.
Aggarwal H, Aggarwal A, Hunter WJ 3rd, Yohannes P, Khan AU, Agrawal DK. Expression of leukemia/lymphoma related factor (LRF/Pokemon) in human benign prostate hyperplasia and prostate cancer. Exp Mol Pathol. 2011; 90(2):226-30.
Cui J, Yang Y, Zhang C, Hu P, Kan W, Bai X, Liu X, Song H. FBI-1 functions as a novel AR co-repressor in prostate cancer cells. Cell Mol Life Sci. 2011;68(6):1091-103.
Kukita A, Kukita T, Nagata K, Teramachi J, Li YJ, Yoshida H, Miyamoto H, Gay S, Pessler F, Shobuike T. The transcription factor FBI-1/OCZF/LRF is expressed in osteoclasts and regulates RANKL-induced osteoclast formation in vitro and in vivo. Arthritis Rheum. 2011;63(9):2744-54.
Zhang QL, Tian DA, Xu XJ. Depletion of Pokemon gene inhibits hepatocellular carcinoma cell growth through inhibition of H-ras. Onkologie. 2011; 34(10):526-31.
Zhao X, Ning Q, Sun X, Tian D. Pokemon reduces Bcl-2 expression through NF-κ Bp65: A possible mechanism of hepatocellular carcinoma. Asian Pac J Trop Med. 2011; 4(6):492-7.
Zu X, Ma J, Liu, F, Tan C, Yu L, Wang J, Cao D, Jiang Y. Pro-oncogene Pokemon promotes breast cancer progression by upregulating survivin expression. Breast Cancer Research 2011; 13:R 26.
Fang F, Yang L, Tao Y, Qin W. FBI-1 promotes cell proliferation and enhances resistance to chemotherapy of hepatocellular carcinoma in vitro and in vivo. Cancer. 2012;118(1):134-46.
Lin CC, Zhou JP, Liu YP, Liu JJ, Yang XN, Jazag A, Zhang ZP, Guleng B, Ren JL. The silencing of Pokemon attenuates the proliferation of hepatocellular carcinoma cells in vitro and in vivo by inhibiting the PI3K/Akt pathway. PLoS One. 2012; 7(12):e51916.
Liu K, Liu F, Zhang N, Liu S, Jiang Y. Pokemon Silencing Leads to Bim-Mediated Anoikis of Human Hepatoma Cell QGY7703. Int J Mol Sci. 2012; 13(5):5818-31.
Yuan B, Xian R, Wu X, Jing J, Chen K, Liu G, Zhou Z. Endoplasmic reticulum chaperone glucose regulated protein 170-Pokemon complexes elicit a robust antitumor immune response in vivo. Immunobiology. 2012; 217(7):738-42.
Yuan B, Zhao L, Xian R, Zhao G. Identification of novel HLA-A(*)0201-restricted CTL epitopes from Pokemon. Cell Immunol. 2012; 274(1-2):54-60.
Jin XL, Sun QS, Liu F, Yang HW, Liu M, Liu HX, Xu W, Jiang YY. microRNA 21-mediated suppression of sprouty1 by Pokemon affects liver cancer cell growth and proliferation. J Cell Biochem. 2013; 114(7):1625-33.
Li Y, Xu S, Wang X, Shi H, Sun Z, Yang Z. Tumor-specific RNA interference targeting Pokemon suppresses tumor growth and induces apoptosis in prostate cancer. Urology. 2013; 81(2):467.e1-7.
Lunardi A, Guarnerio J, Wang G, Maeda T, Pandolfi PP. Role of LRF/Pokemon in lineage fate decisions. Blood. 2013; 121(15):2845-53.
We Also Recommend