Western blot analysis of E2F-1 was performed by loading 30 ug of HeLa (lane1), Hep G2 (lane2), A431 (lane3), U2OS (lane4), HCT 116 (lane5) cell tissue lysate using Novex® NuPAGE® 12 % Bis-Tris gel (NP0342BOX), XCell SureLock™ Electrophoresis System (EI0002), Novex® Sharp Pre-Stained Protein Standard (LC5800), and iBlot® Dry Blotting System (IB21001). Proteins were transferred to a nitrocellulose membrane and blocked with 5 % skim milk at 4°C overnight. E2F-1 was detected at ~47 and 60 kDa using E2F-1 Mouse Monoclonal Antibody (321400) at 1-3 ug/ml in 5 % skim milk for 3 hour at room temperature on a rocking platform. Goat Anti-Mouse - HRP Secondary Antibody (626520) at 1:4000 dilution was used and chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (WP20005).
|Tested species reactivity||Human, Mouse, Rat|
|Published species reactivity||Human, Mouse, Not Applicable|
|Host / Isotype||Mouse / IgG2a|
|Immunogen||Recombinant human E2F-1 protein.|
|Storage buffer||PBS, pH 7.4|
|Contains||0.1% sodium azide|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/million cells|
|Immunohistochemistry (IHC)||0.1-1.0 ug/ml|
|Immunoprecipitation (IP)||2-5 ug|
|Western Blot (WB)||1-3 µg/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
E2F1 is a member of the E2F family of transcription factors. The E2F family plays a crucial role in the control of cell cycle and action of tumor suppressor proteins and is also a target of the transforming proteins of small DNA tumor viruses. The E2F proteins contain several evolutionally conserved domains found in most members of the family. These domains include a DNA binding domain, a dimerization domain which determines interaction with the differentiation regulated transcription factor proteins (DP), a transactivation domain enriched in acidic amino acids, and a tumor suppressor protein association domain which is embedded within the transactivation domain. This protein and another 2 members, E2F2 and E2F3, have an additional cyclin binding domain. This protein binds preferentially to retinoblastoma protein pRB in a cell-cycle dependent manner. It can mediate both cell proliferation and p53-dependent/independent apoptosis.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Triptolide abrogates growth of colon cancer and induces cell cycle arrest by inhibiting transcriptional activation of E2F.
32-1400 was used in immunoprecipitation to study the effect of triptolide on colon cancer growth and cell cycle arrest and its mechanism.
|Oliveira AR,Beyer G,Chugh R,Skube SJ,Majumder K,Banerjee S,Sangwan V,Li L,Dawra RK,Subramanian S,Saluja AK,Dudeja V||Laboratory investigation; a journal of technical methods and pathology (95:648)||2015|
Genomic portrait of resectable hepatocellular carcinomas: implications of RB1 and FGF19 aberrations for patient stratification.
32-1400 was used in immunohistochemistry - paraffin section to identify the prognostic markers of resectable hepatocellular carcinomas
|Ahn SM,Jang SJ,Shim JH,Kim D,Hong SM,Sung CO,Baek D,Haq F,Ansari AA,Lee SY,Chun SM,Choi S,Choi HJ,Kim J,Kim S,Hwang S,Lee YJ,Lee JE,Jung WR,Jang HY,Yang E,Sung WK,Lee NP,Mao M,Lee C,Zucman-Rossi J,Yu E,Lee HC,Kong G||Hepatology (Baltimore, Md.) (60:1972)||2014|
Repair of oxidative DNA damage, cell-cycle regulation and neuronal death may influence the clinical manifestation of Alzheimer's disease.
32-1400 was used in immunohistochemistry - paraffin section to study the clinical manifestation of Alzheimer's disease involving cell-cycle regulation, neuronal death and repair of oxidative DNA damage
|Silva AR,Santos AC,Farfel JM,Grinberg LT,Ferretti RE,Campos AH,Cunha IW,Begnami MD,Rocha RM,Carraro DM,de Bragança Pereira CA,Jacob-Filho W,Brentani H||PloS one (9:null)||2014|
Identification of prognostic biomarkers for glioblastomas using protein expression profiling.
32-1400 was used in immunohistochemistry - paraffin section to identify diagnostic and prognostic markers for glioblastoma
|Jung Y,Joo KM,Seong DH,Choi YL,Kong DS,Kim Y,Kim MH,Jin J,Suh YL,Seol HJ,Shin CS,Lee JI,Kim JH,Song SY,Nam DH||International journal of oncology (40:1122)||2012|
Impact of E2F-1 expression on clinical outcome of gastric adenocarcinoma patients with adjuvant chemoradiation therapy.
32-1400 was used in immunohistochemistry - paraffin section to assess expression and prognostic significance of E2F-1 and thymidylate synthase in R(0)-resected gastric adenocarcinoma patients
|Lee J,Park CK,Park JO,Lim T,Park YS,Lim HY,Lee I,Sohn TS,Noh JH,Heo JS,Kim S,Lim DH,Kim KM,Kang WK||Clinical cancer research : an official journal of the American Association for Cancer Research (14:82)||2008|
|Not Applicable||Not Cited||
Histone deacetylase activity in conjunction with E2F-1 and p53 regulates Apaf-1 expression in 661W cells and the retina.
32-1400 was used in ChIP assay and western blot to elucidate the molecular mechanism of HDAC-mediated regulation of Apaf-1
|Wallace DM,Cotter TG||Journal of neuroscience research (87:887)||2009|
|Not Applicable||Not Cited||
Overexpression of N-Myc rapidly causes acute myeloid leukemia in mice.
32-1400 was used in western blot to determine whether N-Myc overexpression is highly oncogenic in mouse myeloid cells
|Kawagoe H,Kandilci A,Kranenburg TA,Grosveld GC||Cancer research (67:10677)||2007|
|The novel ETS factor TEL2 cooperates with Myc in B lymphomagenesis.||Cardone M,Kandilci A,Carella C,Nilsson JA,Brennan JA,Sirma S,Ozbek U,Boyd K,Cleveland JL,Grosveld GC||Molecular and cellular biology (25:2395)||2005|