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Also for E2F1 (NM_005225)
|This affinity purified antibody was prepared from whole rabbit serum produced by repeated immunizations with a synthetic peptide corresponding to amino acids 360-369 of Human E2F-1.|
||ELISA: 1:20,000 - 1:100,000, WB: 1:250 - 1:2,000, IHC: 2 mg/ml - 20 ug/ml
|0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2|
|E2F-1 (also known as tran-scription factor E2F-1, Retinoblastoma binding protein 3, RBBP-3, PRB-binding protein E2F-1, PBR3, Retinoblastoma-associated protein 1 and RBAP-1) is a transcription activator that binds DNA cooperatively with DP proteins through the E2 recognition site, 5'-TTTC[CG]CGC-3'. 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 domains con-served through evolution that are found in most members of the family. These domains include a DNA binding domain, a dimerization domain that
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WB using E2F-1 pS364 ab shows detection of a ~47 kDa band corresponding to phosphorylated E2F-1 in induced cell lysates. Panel A shows reactivity using a control ab reactive to all forms of E2F (arrowheads). Panel B shows specific reactivity against phosphorylated E2F-1 (arrowheads) using anti-E2F-1 pS364. Lysates are as follows: CRE/E2F-1 are CRE cells derived from mouse NIH3T3 line transfected with human E2F-1, NIH-3T3 used as a negative control, and MDA-MB-231 cells are a human breast cancer line. Lysate was prepared from untreated cells or cells treated with 2 uM Doxorubicin used as DNA damaging agent. MDA-MB-231 cells were also treated with genistein, a mild DNA damaging agent. The figure shows the same membrane first probed with the anti-E2F-1 pS364 at 1:250, then stripped and re-probed with the pan E2F ab used as a positive control. The positive control ab shows an E2F-1 band in all human cell lines, but not mouse cells. Treatment with doxorubicin increases the expression of E2F-1 as shown in Panel A. After film development, images were overlapped to confirm that specific anti-E2F-1 pS364 staining shown treated human cells in Panel B specifically aligns with E2F-1 staining shown in Panel A.
Anti- E2F-1 pS364 antibody was used at a 10 µg/ml to detect nuclear and occasion-ally cytoplasmic signal in a variety of tissues including multi-human, multi-brain and multi-cancer slides. Within the multi-tumor block, the antibody showed variable levels of nuclear staining between individual tumors, with some tumors showing strong staining. This image shows E2F-1 pS364 staining of human breast carcinoma. Tissue was formalin-fixed and paraffin embedded.