mAbs Against HUMAN Protein
E4F1
| STATUS: |  |
|---|---|
| CONTACT INFORMATION: | Monoclonal Antibodies Unit, Centro Nacional de Investigaciones Oncológicas |
| STATUS: | Validated |
| TYPE: | mouse monoclonal |
| CLONE NAME: | 272G |
| PROTEIN: | Transcription factor E4F1 |
| PROTEIN WEB: | https://www.uniprot.org/uniprot/Q66K89 |
| ANTIGEN USED: | GST-E4F1 recombinant protein |
| FUSION PARTNER: | NS1/Ag4-1 (NS1) cells |
| ISOTYPE: | IgG1 |
| SPECIES REACTIVITY: | Human |
| PREPARATION AND STORAGE: | Aliquot and store at 4C. Do not freeze |
Description
May function as a transcriptional repressor. May also function as a ubiquitin ligase mediating ubiquitination of chromatin-associated TP53. Functions in cell survival and proliferation through control of the cell cycle. Functions in the p53 and pRB tumor suppressor pathways and regulates the cyclin CCNA2 transcription.
Identified as a cellular target of the adenoviral oncoprotein E1A, it is required for both transcriptional activation and repression of viral genes.
Applications
| IHC Techniques | Clone | Dilution | Antibody concentration | Antigen retrieval method | Visualization kit | +/- control | Protein localization | Positivity in other species | Protocol |
|---|---|---|---|---|---|---|---|---|---|
| Frozen tissue and cytospins | |||||||||
Recommended Result obtained is satisfactory. The reagent can be use in this application | 272G | Neat | supernatant | / | |||||
| Paraffin tissue | |||||||||
Recommended Result obtained is satisfactory. The reagent can be use in this application | 272G | 1:15 | supernatant | Tris-EDTA | Novolink | Thymus / | nuclear | Available | |
| Immunofluorescence | |||||||||
Enlarge image
- 272G mAb is able to detect human E4F1 protein in immunocytochemistry
- To confirm that 272G mAb recognizes human E4F1 protein, immunocytochemistry on frozen cytospin preparations of HA-tagged E4F1 expressed in HEK293T was performed. Cytospin preparation of Psf1 transfected cells was used as negative control.
Enlarge image
- 272G mAb can be used to detect E4F1 protein in human frozen tissues.
Enlarge image
- 272G mAb can be used to detect E4F1 protein in human paraffin tissues
| WB Techniques | Clone | Dilution | Antibody concentration | +/- control | Expected MW | Observed Mw | Positivity in other species | Protocol |
|---|---|---|---|---|---|---|---|---|
| Western Blotting | ||||||||
Recommended Result obtained is satisfactory. The reagent can be use in this application | 272G | Neat | supernatant | Jurkat cell line / | 83kDa | 83kDa | Available | |
| Immunoprecipitation | ||||||||
Enlarge image
- 272G mAb is able to detect human E4F1 protein by WB.
- LANES
Lane 1 HEK-E4F1-HA (10ug) (+)
Lane 2 HEK-mCD8A-MYC (10ug) (-)
Lane 3 Human tonsil (100ug) (-)
Lane 4 253J cell line (100ug) (-)
Lane 5 U251MG cell line (100ug) (-)
Lane 6 Jurkat cell line (100ug) (+)
Lane 7 MCF7 cell line (100ug) (+)
Lane 8 U87MG cell line (100ug) (-)
Lane 9 NIH-3T3 cell line (100ug) (-)
Anti-GAPDH was used as loading control