Mouse anti-Human / Mouse / Rat p73 (deltaNp73) - IMG-313A

Background
p73 was identified as a long-lost cousin of tumor suppressor protein, p53 (1). p73 has high homology with p53 as well as with p63, a gene implicated in the maintenance of epithelial stem cells. Significant homology between p53, p63, and p73 (approximately 63% amino acid identity in the DNA-binding domain suggest that they may have overlapping functions in the regulation of gene expression (2). The targeted disruption of p73 gene leads to defects hippocampal dysgenesis, hydrocephalus, chronic inflammation and infections (3). Recently, spilicing variant mRNAs of p73 has been identified in MCF-7, a breast carcinoma cell line. These mRNAs code for variant p73 proteins bearing distinct carboxy-terminal structures suggesting that the carboxy-terminal region of p73 may be important for the functions of this protein (4). Recently, Yang et al. (2000) have cloned a p73 variant that lacks an amino-terminal transactivation domain, called deltaNp73 (5). The dNp73 message is transcribed from an alternative promoter located in intron 3.

Antigen
This antibody was developed against a peptide corresponding to amino acid residues 2-13 LYVGDPARHLAT of human dNp73. Mouse and human sequences are more than 95 % identical at these amino acid residues. In mouse, alanine at position 8 is replaced by methionine.

Application Notes
1. IMG-313A reacts with an epitope that is located on the N-terminal region of human dNp73 protein. It does not cross react with full-length p73.
2. IMG-313A (clone 38C674.2) is also referred to as OP181 or OP181, clone 38C674 in the literature.
3. 7. Caspase-cleaved p73 fragments, including 55 kDa and 45 kDa, have been described (Sayan et al, 2008). The ability of a given antibody to recognize a p73 fragment will depend on whether or not the antibody epitope is contained in the fragment. As p73 research continues to evolve, researchers are encouraged to refer to the published literature using the p73 antibodies to gain additional information.

Genebank Info (Protein)
NP_005418

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Reference
1. Ueda Y, Hijikata M, Takagi S, Chiba T, and Shimotohno K. Oncogene 18: 4993-4998 (1999).
2. Yang A, Walker N, Bronson R, Kaghad M, Oosterwegel M, et al. Nature 404: 99-103 (2000).
3. Levrero M, De Laurenzi V, Costanz0 A, Gong J, Wang JY, and Melino G. J Cell Sci 113 (pt 10): 1661-1670 (2000).
4. Kaghad M, Bonnet H, Yang A, Creancier L, Biscan JC., et al. Cell 90: 809-819 (1997).
5. Sayan AE, BS Sayan, V Gogvadze, D Dinsdale, U Nyman, TM Hansen, B Zhivotovsky, GM Cohen, RA Knight, G Melino. p73 and caspase-cleaved p73 fragments localize to mitochondria and augment TRAIL-induced apoptosis. Oncogene 27:4363-4372 (2008).

Product Citations
1. The p73 Gene Is an Anti-Tumoral Target of the RARb/g-Selective Retinoid Tazarotene. Marina Papoutsaki, Mauro Lanza,
Barbara Marinari,
Steven Nistico, Francesca Moretti, Massimo Levrero, Sergio Chimenti,
and Antonio Costanzo. J Invest Dermatol. 123:1162 –1168 (2004). IMGENEX antibodies cited: 1.  p73 (IMG-259A) [WB, Fig 1AB, (immortalized keratinocytes C5N and HaCaT cell lines), Fig 4A (HaCaT cell line) ]. 2. deltaNp73 (IMG-313A) [WB, Fig 1AB (C5N and HaCat cell lines), Fig 4A (HaCaT cell line) ].

2. DN-p73 is activated after DNA damage in a p53-dependent manner to regulate p53-induced cell cycle arrest. Stefania Vossio, Emanuele Palescandolo, Natalia Pediconi, Francesca Moretti, Clara Balsano, Massimo Levrero, Antonio Costanzo. Oncogene 21, 3796-3803 (2002). IMGENEX antibodies cited: 1.  deltaNp73(IMG-313A)  [WB, Fig 2B, Fig 4B (human osteosarcoma U2OS cells) ]. 
3. Treatment with arsenic trioxide (ATO) and MEK1 inhibitor activates the p73-p53AIP1 apoptotic pathway in leukemia cells. Paolo Lunghi, Antonio Costanzo, Massimo Levrero, and Antonio Bonati. Katarina Tomkova, Abbes Belkhiri, Wael El-Rifai, and Alexander I. Zaika. Blood, 104: 519-525 (2004). IMGENEX antibodies cited: 1. p73 (IMG-259A) [WB, Fig 1CD, Fig 2B (NB4 acute promyelocytic and K562 erythroleukemia cells) ].  2. deltaNp73 (IMG-313A)  [WB, Fig 1CD, Fig 2B (NB4 and K562 cells) ].

4. p73 Isoforms Can Induce T-Cell Factor–Dependent Transcription in Gastrointestinal Cells. Katarina Tomkova, Abbes Belkhiri, Wael El-Rifai, and Alexander I. Zaika. Cancer Res., 64: 6390-6393 (2004). IMGENEX antibodies cited: deltaNp73 (IMG-313A) [WB, no data present (AGS gastric cell line) ].

5. Spatiotemporal switch from deltaNp73 to TAp73 isoforms during nephrogenesis: Impact on differentiation gene expression. Zubaida Saifudeen, Virginia Diavolitsis, Jana Stefkova, Susana Dipp, Hao Fan, and Samir S. El-Dahr. J. Biol. Chem., 280: 23094-23102 (2005). IMGENEX anitbodies cited: 1. deltaNp73 (IM-313A) [IHC (paraffin), Fig 5 (adult rat kindey) ], [WB, Fig 5 (adult rat kidney) ]. 
6. MEK1 inhibition sensitizes primary acute myelogenous leukemia to arsenic trioxide-induced apoptosis. Paolo Lunghi, Antonio Costanzo, Luigi Salvatore, Nelida Noguera, Laura Mazzera, Antonio Tabilio, Francesco Lo-Coco, Massimo Levrero, and Antonio Bonati. Blood, Feb 2006; 10.1182/blood-2005-07-2829.    
7. p53 dependent and independent sensitivity to oxaliplatin of colon cancer cells. Florent Toscano, Béatrice Parmentier, Zineb El Fajoui, Yann Estornes, Jean-Alain Chayvialle, Jean-Christophe Saurin and Jacques Abello. Biochemical Pharmacology, 74 (3): 392-406 (2007). IMGENEX antibodies cited: deltaNp73 (IMG-313A) [WB, Fig 9 (oxaliplatin treated HCT116, Isreco1, and V9P cell lines) ].
8. Acetylcholinesterase-R increases germ cell apoptosis but enhances sperm motility. Mor I, E Sklan, E Podoly, M Pick, M Kirschner, L Yogev, S Itach, L Schreiber, B Geyer, T Mor, D Grisaru. Journal of Cellular and Molecular Medicine doi: 10.1111/j.1582-4934.2008.00231.x (2008). Imgenex products cited [WB (testicular tissue from male transgenic (TGR) mice)]:
1. IMG-246 (TAp73) Fig. 3D
2. IMG-313 (ΔNp73) Fig. 3D
Note: IMG-246 and IMG-314 detected p73 forms  in whole cell lysate as well as p73 forms that co-immunoprecipitated with RACK1, Fig  3D
9. Cooperation between p53 and p73 in cisplatin-induced apoptosis in ovarian carcinoma cells. Righetti S, Perego P, Carenini N, Zunino F. Cancer Letters 263: 140-144 (2008). WB (human ovarian cell lines A2780 and A2780/BBR3464), Fig. 3.
10. New antibodies recognizing p73: Comparison with commercial antibodies. Sayan AE, A Paradisi, B Vojtesek, RA Knight, G Melino, E Candi. Biochemical Biophysical Res Communications. 330:186-193. (2005). IMGENEX antibodies cited (p73 transfected cells, SK-N-SH human neuroblastoma cells, HaCaT human keratinocyte cells):
1. p73 (IMG-246): IP, Fig 3B (p73 transfected cells); WB, Figs 1 and 2 (p73 transfected cells).
2. p73 (IMG-259A): WB, Figs 1 and 2 (p73 transfected cells)
3. p73 (IMG-313A): WB, Figs 1, 2 and 3 (p73 transfected cells)

Note: See Table 1 for a summary of antibody results (WB and IF/ICC) in p73 transfected cells, SK-N-SH cells and HaCaT cells) IMG-313A (Clone 38C674.2) is referred to as OP181 in this publication.
11. Delta TAp73 upregulation correlates with poor prognosis in human tumors: putative in vivo network involving p73 isoforms, p53 and E2F-1. Dominguez G, JM Garcia, C Pena, J Silva, V Garcia, L Martinez, C Maximiano, ME Gomez, JA Rivera, C Garcia-Andrade and F Bonilla. J Clinical Oncology 24:1-11 (2006). Imgenex antibodies cited  [IHC (F) in human colon and breast tumors], Fig 2:
1. p73 (IMG-313A)
2. p73 (IMG-246)
12. TAp73 knockout shows genomic instability with infertility and tumor suppressor functions. Tomasini R, K Tsuchihar, M Wilhelm, M Fuitani, A Rufini, C Cheung, F Khan, A Itie-Youten, A Wakeham, M Tsao, J Iovanna, J Squire, I Jurisica, D Kaplan, G Melino, A Jurisicova, T Mak. Genes and Development 22:2677-2691 (2008). WB (mouse embryonic fibroblasts), Fig. 1D.
13. Targeting MEK/MAPK signal transduction module potentiates ATO-induced apoptosis in multiple myeloma cells through multiple signaling pathways. Lungli P, N Giuliani, L Mazzera, G Lombardi, M Ricca, A Corradi, A Cantoni, L Salbatore, R Riccioni, A Constanzo, U Testa, M Levrero, V Rizzoli, A Bonati. Blood 112: 2450-2462. Imgenex antibodies cited:
1. p73 (IMG-246): WB (human myeloma cell lines (HMCLs) XG-6 and XG-1), Fig. 4Ci.
2. p73 (deltaNp73) [IMG-313A]: WB (HMCLs), Fig. 4Ai.
3. DcR2 (IMG-121): WB (HMCLs), Fig. 5A.

14. STI571 prevents apoptosis, tau phosphorylation and behavioural impairments induced by Alzheimer's B-amyloid deposits. Cancino G, E Toledo, N Leal, D Hernandez, F Yevenes, N Inestrosa, A Alvarez. Brain 131: 2425-2442 (2008).  Imgenex antibodies cited [WB (rat hippocampal cells)]:
1. IMG-246 (p73): Fig. 5A.
2. IMG-313 (ΔNp73): Fig. 5B.
15. p73 plays a role in erythroid differentiation through GATA1 induction. Marques-Gacia F, N Ferrandiz, R Fernandez-Alonso, L Gonzalez-Cano, M Herreros-Villanueva, M Rosa-Garrido, B Fernandez-Garcia, J P Vaque, M M Marques, M E Alonso, J C Segovia, J Leon, M C Marin. JBC 284: 21139- 21156 (2009). IMGENEX antibodies cited for ChIP in K562 cells (Figs 4C, 6) and fetal liver extracts from mouse embryos (Fig 10):
1. p73 (IMG-246) 
2. p73 [deltaNp73 (IMG-313)]
16. Interaction of the p53 protein family in cellular stress response in gastrointestinal tumors. Vilgelm A, M Washington, J Wei, H Chen, V Prassolov, A Zaika. Mol Cancer Ther. 9: 693-705 (2010). IMGENEX products cited:
1. delta Np73 (IMG-313A) IHC (paraffin), Fig 1A, human colorectal adenocarcinoma cells.
2. NOXA (IMG-349A) WB, Fig 2A, Fig 2B, Fig 2D, colon adenocarcinoma and esophageal carcinoma cell lines.
17.

p73 and p53 pathway in human breast cancers. Bozzetti C, R Nizzoli, A Musolino, EM Martella, P Crafa, CA Lagrasta, R Camisa, A Bonati, P Lunghi, A Ardizzoni. JCO 25:1451-1453. IMGENEX antibodies cited (human primary breast cancer cytospins, NB4 and K562 cell lines), ICC:

1. p73 mAb, clone 5B429 (IMG-246), Table 1

2. p73 (deltaNp73) mAb, clone 38C674.2 (IMG-313A), Fig 1, Table 1 

Note:  p73 expression was also confirmed in the cancer cells and cell lines by western blot using IMG-313A and IMG-246. 
18. Isoform-specific p73 knockout mice reveal a novel role for deltaNp73 in the DNA damage response pathway. Wilhelm MT, A Rufini, MK Wetzel, K Tshuchihara, S Inoue, R Tomasini, A Itie-Youten, A Wakeham, M Arsenian-Henriksson, G Melino, DR Kaplan, FD Miller, TW Mak. Genes & Development 24:549-560 (2010). WB (SH-Sy5Y human neuroblastoma cells): Fig 7B. Lysates were IP'd with a 53BP1 antibody, and immunoprecipitates were western blotted with the deltaNp73 (IMG-313A) antibody. The results showed that deltaNp73 coIP'd with 53BP1. 
19. Multiple isoforms of the tumor protein p73 are expressed in the adult human telencephalon and choroid plexus and present in the cerebrospinal fluid. Cabrera-Socorro A, MP Morlans, MLS Sola, FJG Delgado, A Castaneyra-Perdomo, MC Marin, G Meyer. European J Neuroscience 23:2109-2118 (2006). IHC (paraffin): human brain (Figs 2D, 3C, 3D), adult mouse brain (Fig 2F), human CSF (Figs 4B, 4C).
20. I kappa B kinase beta promotes cell survival by antagonizing p53 functions through delta Np73 alpha phosphorylation and stabilization. Accardi R, M Scalise, T Gheit, I Hussain, J Yue, C Carreira, A Collino, C Indiveri, L Gissmann, B Sylla, M Tommasino. Mol Cell Biol doi:10.1128/MCB.00964-10 (2011). IMGENEX product cited: p73 deltaNp73 (IMG-313A).
1. WB: Human primary keratinocytes, Fig 1A.
2. WB: HEK293 cells, Fig 2C.
21. Alternate AChE-R variants facilitate cellular metabolic activity and resistance to genotoxic stress through enolase and RACK1 interactions. Mor I, T Bruck, D Greenberg, A Berson, L Schreiber, D Grisaru, H Soreq. Chemico-Biological Interactions 175:11-21 (2008). IMGENEX products cited: deltaNp73 (IMG-313A), tankyrase (IMG-146). WB: GFP expressing CHO cell line, Fig 3E.
22. p73 expression in medulloblastoma: TAp73/deltaNp73 transcript detection and possible association of p73alpha/deltaNp73 immunoreactivity with survival. Zitterbart K, I Zavrelova, J Kadlecova, R Spesna, A Kratochvilova, Z Pavelka, J Sterba. Acta Neuropathol 114:641-650 (2007). IMGENEX product cited: deltaNp73 (IMG-313A). IHC (paraffin): Human medulloblastoma tissue, Fig 2 & Fig 3.
23.