ET1701-73

E2F1 Recombinant Rabbit Monoclonal Antibody [JJ092-02]

cat.: ET1701-73

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IF-Cell, IF-Tissue, IHC-P, IP, FC
Clonality:	Monoclonal
Clone number:	JJ092-02

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	1ug/ul
Purification:	Protein A affinity purified.
Molecular weight:	Predicted band size: 47 kDa
Isotype:	IgG
Immunogen:	Synthetic peptide within Human E2F1 aa 371-404 / 437.
Positive control:	HT-29 cell lysate, SW480 cell lysate, SW620 cell lysate, HepG2 cell lysate, NIH/3T3 cell lysate, rat heart tissue lysate, Hela, MCF-7, HepG2, human tonsil tissue, mouse pancreas tissue, rat heart tissue.
Subcellular location:	Nucleus.
Recommended Dilutions: WB IF-Cell IF-Tissue IHC-P FC IP	1:500-1:10,000 1:100-1:500 1:100-1:500 1:1,000-1:5,000 1:1,000 1-2μg/sample
Uniprot #:	SwissProt: Q01094 Human \| Q61501 Mouse \| O09139 Rat
Alternative names:	DmelCG6376 Dmel_CG6376 drosE2F1 E(Sev-CycE)3A E(var)3-93E E2-promoter binding facto E2F 1 E2F transcription factor 1 E2F-1 E2f-PA E2f-PB E2f-PC E2F1 E2f1 E2F transcription factor 1 E2F1_HUMAN Evar(3)164 KIAA4009 l(3)07172 l(3)j3B1 l(3)j3C2 l(3)rM729 mKIAA4009 OTTHUMP00000030661 PBR3 PRB binding protein E2F 1 PRB-binding protein E2F-1 RBAP 1 RBAP-1 RBAP1 RBBP-3 RBBP3 RBP 3 RBP3 Retinoblastoma-associated protein 1 Retinoblastoma-binding protein 3 Transcription factor E2F1

Images

Fig1: Western blot analysis of E2F1 on different lysates with Rabbit anti-E2F1 antibody (ET1701-73) at 1/500 dilution.

Lane 1: HT-29 cell lysate
Lane 2: SW480 cell lysate
Lane 3: SW620 cell lysate
Lane 4: HepG2 cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 47 kDa
Observed band size: 60 kDa

Exposure time: 3 minutes; ECL: K1801;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1701-73) at 1/500 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

Fig2: Western blot analysis of E2F1 on different lysates with Rabbit anti-E2F1 antibody (ET1701-73) at 1/1,000 dilution.

Lane 1: NIH/3T3 cell lysate (10 µg/Lane)
Lane 2: Rat heart tissue lysate (20 µg/Lane)

Predicted band size: 47 kDa
Observed band size: 60 kDa

Exposure time: 1 minute 2 seconds; ECL: K1801;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1701-73) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

	Fig3: 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).
	Fig4: 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).
	Fig5: 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).
	Fig6: Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Rabbit anti-E2F1 antibody (ET1701-73) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1701-73) at 1/1,000 dilution for 1 hour 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.

	Fig7: Immunohistochemical analysis of paraffin-embedded mouse pancreas tissue with Rabbit anti-E2F1 antibody (ET1701-73) at 1/5,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1701-73) at 1/5,000 dilution for 1 hour 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.
	Fig8: Immunohistochemical analysis of paraffin-embedded rat heart tissue with Rabbit anti-E2F1 antibody (ET1701-73) at 1/5,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1701-73) at 1/5,000 dilution for 1 hour 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.
	Fig9: Flow cytometric analysis of HepG2 cells labeling E2F1. Cells were fixed and permeabilized. Then stained with the primary antibody (ET1701-73, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
	Fig10: Flow cytometric analysis of NIH/3T3 cells labeling E2F1. Cells were fixed and permeabilized. Then stained with the primary antibody (ET1701-73, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

Fig11: E2F1 was immunoprecipitated from 0.2 mg HepG2 cell lysate with ET1701-73 at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using ET1701-73 at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: HepG2 cell lysate (input)
Lane 2: ET1701-73 IP in HepG2 cell lysate
Lane 3: Rabbit IgG instead of ET1701-73 in HepG2 cell lysate

Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 26 seconds; ECL: K1801

Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.