E2F Transcription Factor 1 (E2F1) (pS364) Antikörper

Antigen: E2F Transcription Factor 1 (E2F1)

Synonyme: RBP3, E2F-1, RBAP1, RBBP3, KIAA4009, mKIAA4009, E2F1, xE2F

Epitope: pS364

Klonalität: Polyklonal

Wirt: Kaninchen

Reaktivität: Human, Rind (Kuh), Maus, Ratte (Rattus)

Applikation: Immunhistochemie (IHC), ELISA, Western Blot (WB), Immunpräzipitation (IP)

Produktnummer: ABIN129619

Produktbeschreibung

Weitere Bezeichnung: E2F-1 pS364

Gen-ID: NP_005216

Immunogen: 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.

Format: Liquid (sterile filtered)

Beschreibung: 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 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 as well as E2F2 and E2F3 have an additional cyclin binding domain. This protein binds preferentially to retinoblastoma protein pRB in a cell-cycle dependent manner and mediates both cell proliferation and p53-dependent/independent apoptosis. Increased nuclear expression of this protein has been reported in a variety of cancers .

Spezifität: 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.

Anwendungen

Applikationshinweise: Recommended Dilutions: ELISA 1:20,000 - 1:100,000 WESTERN BLOT 1:250 - 1:2,000 IMMUNOHISTOCHEMISTRY 2 ug/ml - 20 ug/ml OTHER APPLICATIONS User Optimized. 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.

Konzentration: 0.80 mg/ml (by UV absorbance at 280 nm)

Buffer: Stabilizer: None. Preservative: 0.01% (w/v) Sodium Azide. Buffer: 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2.

Lagerung: Store vial at -20° C prior to opening. Dilute only prior to immediate use. For extended storage aliquot contents and freeze at -20° C or below. Avoid cycles of freezing and thawing. Expiration date is one (1) year from date of opening. Figure. Western blot using 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-E2F1 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 lysatewas 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 Ddamaging agent. The figure shows the same membrane first

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Bilder

Western blot using antibodies-online'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-E2F1 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 lysatewas 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 anti-E2F-1 pS364 antibody used at a 1:250 dilution, then stripped and re-probed with the pan E2F antibodyused as a positive control. The positive control antibody clearlyshows an E2F-1 band in all human cell lines, but not mouse cells. Treatment with doxorubicin increases the expression oE2F-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 ABIN102010 for 45 min at room temperature for ECL detection. Personal Communication, XiaoHe Yang, Univ. Oklahoma.