PRODUCT CODE: ET1701-73

Recombinant E2F1 Monoclonal Antibody (ET1701-73)

  • Recombinant

Applications

  • WB

  • ICC

  • IF

  • IHC-P

  • IP

  • FC

REACTIVITY

  • Human

  • Mouse

  • Rat

Western blot analysis of E2F1 on HepG2 cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1701-73, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
  • Western blot analysis of E2F1 on HepG2 cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1701-73, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
  • ICC staining of E2F1 in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1701-73, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • ICC staining of E2F1 in MCF-7 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1701-73, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • ICC staining of E2F1 in HepG2 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1701-73, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • Immunohistochemical analysis of paraffin-embedded mouse pancreas tissue using anti-E2F1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1701-73, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using anti-E2F1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1701-73, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Flow cytometric analysis of E2F1 was done on Hela cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1701-73, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Western blot analysis of E2F1 on HepG2 cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1701-73, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.

Applications

  • WB

  • ICC

  • IF

  • IHC-P

  • IP

  • FC

REACTIVITY

  • Human

  • Mouse

  • Rat

SPECIFICATIONS

Product Type

Recombinant Rabbit monoclonal primary

Product Name

Recombinant E2F1 Monoclonal Antibody (ET1701-73)

Immunogen

Recombinant protein

Host

Rabbit

Positive Control

HepG2 cell lysates, Hela, MCF-7, HepG2, mouse pancreas tissue, human breast carcinoma tissue, human tonsil tissue, human colon tissue, human pancreas tissue.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

JJ092-02

PROPERTIES

Form

Liquid

Storage Condition

Store at +4C after thawing. Aliquot store at -20C or -80C. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Concentration

1 ug/ul

PURIFICATION

Protein A affinity purified.

MOLECULAR WEIGHT

55 kDa

Isotype

IgG

APPLICATION DILUTION

  • WB

  • 1:1,000

  • ICC/IF

  • 1:100-1:500

  • IHC-P

  • 1:50-1:200

  • FC

  • 1:50-1:100

TARGET

UNIPROT #

PROTEIN NAME

E2F1

SYNONYMS

DmelCG6376 antibody; Dmel_CG6376 antibody; drosE2F1 antibody; E(Sev-CycE)3A antibody; E(var)3-93E antibody; E2-promoter binding facto antibody; E2F 1 antibody; E2F transcription factor 1 antibody; E2F-1 antibody; E2f-PA antibody; E2f-PB antibody; E2f-PC antibody; E2F1 antibody; E2f1 E2F transcription factor 1 antibody; E2F1_HUMAN antibody; Evar(3)164 antibody; KIAA4009 antibody; l(3)07172 antibody; l(3)j3B1 antibody; l(3)j3C2 antibody; l(3)rM729 antibody; mKIAA4009 antibody; OTTHUMP00000030661 antibody; PBR3 antibody; PRB binding protein E2F 1 antibody; PRB-binding protein E2F-1 antibody; RBAP 1 antibody; RBAP-1 antibody; RBAP1 antibody; RBBP-3 antibody; RBBP3 antibody; RBP 3 antibody; RBP3 antibody; Retinoblastoma-associated protein 1 antibody; Retinoblastoma-binding protein 3 antibody; Transcription factor E2F1 antibody

SEQUENCE SIMILARITIES

Belongs to the E2F/DP family.

POST-TRANSLATIONAL MODIFICATION

Phosphorylated by CDK2 and cyclin A-CDK2 in the S-phase. Phosphorylation at Ser-364 by CHEK2 stabilizes E2F1 upon DNA damage and regulates its effect on transcription and apoptosis.; Acetylation stimulates DNA-binding. Enhanced under stress conditions such as DNA damage and inhibited by retinoblastoma protein RB1. Regulated by KAP1/TRIM28 which recruits HDAC1 to E2F1 resulting in deacetylation. Acetylated by P/CAF/KAT2B.

SUBCELLULAR LOCATION

Nucleus.

FUNCTION

The human retinoblastoma gene product appears to play an important role in the negative regulation of cell proliferation. Functional inactivation of Rb can be mediated either through mutation or as a consequence of interaction with DNA tumor virus-encoded proteins. Of all the Rb associations described to date, the identification of a complex between Rb and the transcription factor E2F most directly implicates Rb in regulation of cell proliferation. E2F was originally identified through its role in transcriptional activation of the adenovirus E2 promoter. Sequences homologous to the E2F binding site have been found upstream of a number of genes that encode proteins with putative functions in the G1 and S phases of the cell cycle. E2F-1 is a member of a broader family of transcription regulators including E2F-2, E2F-3, E2F-4, E2F-5, E2F-6 and E2F-7 each of which forms heterodimers with a second protein, DP-1, forming an "active" E2F transcriptional regulatory complex.