Rabbit polyclonal antibody raised against synthetic phosphopeptide of E2F1.
Synthetic phosphopeptide corresponding to residues surrounding S364 of human E2F1.
Reactivity occurs against human E2F-1 pS364 protein and This antibody is specific to the phosphorylated form of the protein. Reactivity with non-phosphorylated human E2F-1 is minimal by ELISA. This antibody does not cross-react with E2F-1 phosphorylated at other sites.
Quality Control Testing:
Antibody Reactive Against Synthetic Peptide.
ELISA (1:20000-1:100000) Western Blot 1:250-1:2000 Immunohistochemistry (2-20 ug/mL) The optimal working dilution should be determined by the end user.
In 20 mM KH2PO4, 150 mM NaCl, pH 7.2 (0.01% sodium azide)
Store at 4°C. For long term storage store at -20°C. Aliquot to avoid repeated freezing and thawing.
This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Western blot using E2F1 (phospho S364) polyclonal antibody (Cat # PAB10004) shows detection of a band at ~47 kDa corresponding to phospho-E2F1 in induced cell lysates. Panel A shows reactivity using a control antibody reactive to all forms of E2F (arrowheads). Panel B shows specific reactivity against phosphorylated E2F-1 (arrowheads) using our Phospho-E2F1 S364 polyclonal antibody. Lysates are as follows : CRE/E2F1 are CRE cells derived from mouse NIH/3T3 line transfected with human E2F1, NIH/3T3 used as a negative control, and MDA-MB-231 cells are a human breast cancer line. As indicated each lysate was prepared from untreated cells and cells treated with 2 uM Doxorubicin used as a DNA damaging agent. In addition the MDA-MB-231 cells were also treated with genistein, a mild Ddamaging agent. The figure shows the same membrane first probed with the E2F1 (phospho S364) polyclonal antibody (Cat # PAB10004) used at a 1 : 250 dilution, then stripped and re-probed with the
Immunohistochemistry of E2F1 (phospho S364) polyclonal antibody (Cat # PAB10004) was used at a 10 ug/mL to detect nuclear and occasion-ally cytoplasmic signal in a variety of tissues in-cluding 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 E2F1 pS364 staining of human breast carcinoma. Tissue was formalin-fixed and paraffin embedded. Personal Communication, Tina Roush, Life Span Biosciences, Seattle, WA.
The protein encoded by this gene 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. [provided by RefSeq
OTTHUMP00000030661,retinoblastoma-associated protein 1