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mAbs Against HUMAN Protein

E4F1

STATUS:Antibody Validated
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
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
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