![Western Blot: E2F-1 [p Ser364] Antibody [NB600-900] - Panel A: Control antibody reactive to all forms of E2F (arrowheads). Panel B: Reactivity with phosphorylated E2F-1 (arrows) using E2F-1 [p Ser364] antibody. Lysate cells: CRE/E2F-1 CRE (human E2F-1 transfected mouse NIH3T3); NIH-3T3 (-ve control), & MDA-MB-231. Each lysate prepared from untreated cells & cells with 2 uM Doxorubicin. MDA-MB-231 also treated with genistein. Same membrane probed with E2F-1 [p Ser364] antibody @ 1:250 dilution, then stripped & re-probed with pan E2F antibody (+ve control). +ve control antibody shows an E2F-1 band in all human cell lines, but not mouse cells. Doxorubicin increases expression of E2F-1 (Panel A). Images overlapped to confirm that specific anti-E2F-1 pS364 staining shown in 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 for 45 min at RT for ECL detection.](http://images.novusbio.com/fullsize/E2F-1-[p-Ser364]-Antibody-Western-Blot-NB600-900-img0007.jpg "Western Blot: E2F-1 [p Ser364] Antibody [NB600-900] - Panel A: Control antibody reactive to all forms of E2F (arrowheads). Panel B: Reactivity with phosphorylated E2F-1 (arrows) using E2F-1 [p Ser364] antibody. Lysate cells: CRE/E2F-1 CRE (human E2F-1 transfected mouse NIH3T3); NIH-3T3 (-ve control), & MDA-MB-231. Each lysate prepared from untreated cells & cells with 2 uM Doxorubicin. MDA-MB-231 also treated with genistein. Same membrane probed with E2F-1 [p Ser364] antibody @ 1:250 dilution, then stripped & re-probed with pan E2F antibody (+ve control). +ve control antibody shows an E2F-1 band in all human cell lines, but not mouse cells. Doxorubicin increases expression of E2F-1 (Panel A). Images overlapped to confirm that specific anti-E2F-1 pS364 staining shown in 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 for 45 min at RT for ECL detection.")
| Reactivity | Hu, MuSpecies Glossary |
| Applications | WB, ELISA, IHC |
| Clonality | Polyclonal |
| Host | Rabbit |
| Conjugate | Unconjugated |
| Description | This affinity purified antibody is directed against the phosphorylated form of human E2F-1 at the [p Ser364] 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 Store vial at -20C prior to opening. Aliquot contents and freeze at -20C 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 4C as an undiluted liquid. Dilute only prior to immediate use. |
| Immunogen | This affinity purified E2F-1 [p Ser364] Antibody was prepared from whole rabbit serum produced by repeated immunizations with a synthetic peptide corresponding to an internal region near amino acids 350-375 of Human E2F-1. (Uniprot: Q01094) |
| Modification | p Ser364 |
| Specificity | This affinity purified antibody is directed against the phosphorylated form of human E2F-1 at the pS364 residue. 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. |
| Isotype | IgG |
| Clonality | Polyclonal |
| Host | Rabbit |
| Gene | E2F1 |
| Purity | Immunogen affinity purified |
| Innovator's Reward | Test in a species/application not listed above to receive a full credit towards a future purchase. |
| Dilutions |
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| Application Notes | 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. |
| Storage | Store at -20C short term. Aliquot and store at -80C long term. Avoid freeze-thaw cycles. |
| Buffer | 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2 |
| Preservative | 0.01% Sodium Azide |
| Purity | Immunogen affinity purified |
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Secondary Antibodies |
Isotype Controls |
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![Immunohistochemistry: E2F-1 [p Ser364] Antibody [NB600-900] - Affinity Purified E2F-1 [p Ser364] antibody was used at a 10 ug/ml to detect nuclear and occasionally 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.](http://images.novusbio.com/fullsize/E2F-1-[p-Ser364]-Antibody-Immunohistochemistry-NB600-900-img0008.jpg "Immunohistochemistry: E2F-1 [p Ser364] Antibody [NB600-900] - Affinity Purified E2F-1 [p Ser364] antibody was used at a 10 ug/ml to detect nuclear and occasionally 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.")
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