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Primary Antibodies  >  Phospho Specific Antibodies

E2F-1 phospho S364 Antibody

Rabbit Polyclonal
NCI Collaboration
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Western blot using Rockland's Affinity Purified anti-E2F-1 pS364 antibody shows detection of a band at ~47 kDa corresponding to phosphorylated E2F-1 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 anti-E2F-1 pS364 antibody.  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.  As indicated each lysate was prepared from untreated cells and cells treated with 2 µM Doxorubicin used as a DNA damaging agent.  In addition the 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 antibody used at a 1:250 dilution, then stripped and re-probed with the pan E2F antibody used as a positive control.  The positive control antibody clearly 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.   Blots can be processed with HRP conjugated Gt-a-Rabbit IgG MX10 611-103-122 for 45 min at room temperature for ECL detection.  Personal Communication, XiaoHe Yang, Univ. Oklahoma.
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100 µg
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E2F-1 phospho S364 Antibody Properties

Anti-E2F-1 pS364 (RABBIT) Antibody - 600-401-690
Target Species
Known Cross Reactivity
human, chimpanzee
ELISA : 1:20,000 - 1:100,000
Western Blot : 1:250 - 1:2,000
Immunohistochemistry: 2 mg/ml - 20 µg/ml
Other Dilution: User Optimized
Physical State
Liquid (sterile filtered)
Shipping Condition
Dry Ice
1.0 mg/mL by UV absorbance at 280 nm
0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
0.01% (w/v) Sodium Azide
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E2F-1 phospho S364 Antibody Description

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
E2F 1 antibody, E2F transcription factor 1 antibody, E2F-1 antibody, E2f1 E2F transcription factor 1 antibody, KIAA4009 antibody, mKIAA4009 antibody, OTTHUMP00000030661 antibody, PBR 3 antibody, PBR3 antibody
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.
Immunogen Type
Post Translational Modification
Storage Condition
Store vial at -20° C prior to opening. Aliquot contents and freeze at -20° C or below for extended storage. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4° C as an undiluted liquid. Dilute only prior to immediate use.
Application Note
This affinity purified antibody has been tested for use in ELISA, immunohistochemistry and by western blot.  Specific conditions for reactivity should be optimized by the end user. Expect a band approximately 47 kDa in size corresponding to phosphorylated E2F-1 by western blotting in the appropriate cell lysate or extract.  Less than 0.5% reactivity is observed against the non-phosphorylated form of the immunizing peptide.  This antibody is phospho specific for pS364 of E2F-1.
This affinity purified antibody is directed against the phosphorylated form of human E2F-1 at the pS364 residue. The product was affinity purified from monospecific antiserum by immunoaffinity purification.  Antiserum was first purified against the phosphorylated form of the immunizing peptide.  The resultant affinity purified antibody was then cross adsorbed against the non-phosphorylated form of the immunizing peptide.  Reactivity occurs against human E2F-1 pS364 protein and the antibody is specific for the phosphorylated form of the protein. Reactivity with non-phosphorylated human E2F-1 is minimal by ELISA. The antibody does not cross-react with E2F-1 phosphorylated at other sites.  A BLAST analysis was used to suggest reactivity with this protein from human and chimpanzee based on 100% homology for the immunogen sequence.  Cross reactivity with E2F-1 homologues from other sources has not been determined.
Disclaimer Note-General
This product is for research use only and is not intended for therapeutic or diagnostic applications. Please contact a technical service representative for more information. All products of animal origin manufactured by Rockland Immunochemicals are derived from starting materials of North American origin. Collection was performed in United States Department of Agriculture (USDA) inspected facilities and all materials have been inspected and certified to be free of disease and suitable for exportation. All properties listed are typical characteristics and are not specifications. All suggestions and data are offered in good faith but without guarantee as conditions and methods of use of our products are beyond our control. All claims must be made within 30 days following the date of delivery. The prospective user must determine the suitability of our materials before adopting them on a commercial scale. Suggested uses of our products are not recommendations to use our products in violation of any patent or as a license under any patent of Rockland Immunochemicals, Inc. If you require a commercial license to use this material and do not have one, then return this material, unopened to: Rockland Inc., P.O. BOX 5199, Limerick, Pennsylvania, USA.
General Reference
Wang,C., Hou,X., Mohapatra,S., Ma,Y., Cress,W.D., Pledger,W.J. and Chen,J. (2005) Activation of p27Kip1 Expression by E2F1. A negative feedback mechanism. J. Biol. Chem. 280 (13), 12339-12343. Joshi,B., Ordonez-Ercan,D., Dasgupta,P. and Chellappan,S. (2005) Induction of human metallothionein 1G promoter by VEGF and heavy metals: differential involvement of E2F and transcription factors. Oncogene 24 (13), 2204-2217. Sdek,P., Ying,H., Zheng,H., Margulis,A., Tang,X., Tian,K. and Xiao,Z.X. (2004) The central acidic domain of MDM2 is critical in inhibition of retinoblastoma-mediated suppression of E2F and cell growth. J. Biol. Chem. 279 (51), 53317-53322.
Specific Reference
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