Western blot analysis was performed on whole cell extracts (30 ug lysate) of HeLa (Lane 1), SH-SY5Y (lane 2), U-87MG (lane 3), MDA-MB-231 (Lane 4),PC-3 (lane 5). The blots were probed with Anti-E2F3 Transcription Factor Mouse Monoclonal Antibody (Product# MA511319, 2 ug/mL) and detected by chemiluminescence Goat anti-Mouse IgG (H+L) Secondary Antibody, HRP conjugate (Product # 626520, 1:4000 dilution). Two bands 37 and 49 kDa band corresponding to E2F3 Transcription Factor was observed across cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 10 % Bis-Tris gel (Product # NP0301BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
|Tested species reactivity||Human|
|Published species reactivity||Rat, Human|
|Host / Isotype||Mouse / IgG2a|
|Immunogen||Recombinant human E2F-3 protein|
|Storage buffer||PBS, pH 7.4, with 0.2% BSA|
|Contains||0.09% sodium azide|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|ChIP assay (ChIP)||2.5 µg/10^6 cells|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Gel Shift (GS)||1mg/mL|
|Immunofluorescence (IF)||Assay Dependent|
|Western Blot (WB)||2-3 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.
MA5-11319 targets E2F-3 Transcription Factor in GS and IF applications and shows reactivity with Human samples.
The MA5-11319 immunogen is recombinant human E2F-3 protein.
E2F transcription factors are functionally regulated by binding to Rb p110, p107, and p130. E2F-3 is regulated by complex formation with Rb p110. E2F family members bind DNA as heterodimers with members of the DP family of polypeptides
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Anti-apoptotic function of the E2F transcription factor 4 (E2F4)/p130, a member of retinoblastoma gene family in cardiac myocytes.
MA5-11319 was used in ChIP assay to study the anti-apoptotic function of the E2F transcription factor-4 in differentiated cardiomyocytes
|Dingar D,Konecny F,Zou J,Sun X,von Harsdorf R||Journal of molecular and cellular cardiology (53:820)||2012|
Inactivation of the Rb pathway and overexpression of both isoforms of E2F3 are obligate events in bladder tumours with 6p22 amplification.
MA5-11319 was used in immunohistochemistry to investigate the role of Rb pathway inaction and E2F3 overexpression in bladder tumours
|Hurst CD,Tomlinson DC,Williams SV,Platt FM,Knowles MA||Oncogene (27:2716)||2008|