HA721192

eIF4E Recombinant Rabbit Monoclonal Antibody [JE60-70]

cat.: HA721192

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IHC-P, IF-Cell, FC
Clonality:	Monoclonal
Clone number:	JE60-70

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃. 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: 25 kDa
Isotype:	IgG
Immunogen:	Recombinant protein within Human eIF4E aa 118-217 / 217.
Positive control:	HL-60 cell lysate, Daudi cell lysate, NIH/3T3 cell lysate, Mouse spleen tissue lysate, Rat spleen tissue lysate, human testis tissue, human colon carcinoma tissue, human breast carcinoma tissue, mouse hippocampus tissue, mouse brain tissue, SK-Br-3.
Subcellular location:	Cytoplasm, P-body, Stress granule, Nucleus.
Recommended Dilutions: WB IHC-P IF-Cell FC	1:500-1:2,000 1:200 1:50 1:500-1:1,000
Uniprot #:	SwissProt: P06730 Human \| P63073 Mouse \| P63074 Rat
Alternative names:	AUTS19 CBP eIF 4E eIF 4F 25 kDa subunit EIF 4F eIF-4E eIF-4F 25 kDa subunit eIF4E EIF4E1 EIF4EL1 EIF4F Eukaryotic translation initiation factor 4 E Eukaryotic translation initiation factor 4E Eukaryotic translation initiation factor 4E like 1 IF4E_HUMAN Messanger RNA Cap Binding Protein eIF 4E MGC111573 mRNA cap binding protein mRNA cap-binding protein

Images

Fig1: Western blot analysis of eIF4E on different lysates with Rabbit anti-eIF4E antibody (HA721192) at 1/500 dilution.

Lane 1: HL-60 cell lysate
Lane 2: Daudi cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 25 kDa
Observed band size: 25 kDa

Exposure time: 2 minutes;

15% 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 (HA721192) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.

Fig2: Western blot analysis of eIF4E on different lysates with Rabbit anti-eIF4E antibody (HA721192) at 1/2,000 dilution.

Lane 1: NIH/3T3 cell lysate
Lane 2: Mouse spleen tissue lysate
Lane 3: Rat spleen tissue lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 25 kDa
Observed band size: 25 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 (HA721192) at 1/2,000 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: Immunohistochemical analysis of paraffin-embedded human testis tissue with Rabbit anti-eIF4E antibody (HA721192) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (HA721192) at 1/200 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.
	Fig4: Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue with Rabbit anti-eIF4E antibody (HA721192) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (HA721192) at 1/200 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.
	Fig5: Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue with Rabbit anti-eIF4E antibody (HA721192) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (HA721192) at 1/200 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.

	Fig6: Immunohistochemical analysis of paraffin-embedded mouse hippocampus tissue with Rabbit anti-eIF4E antibody (HA721192) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (HA721192) at 1/200 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 brain tissue with Rabbit anti-eIF4E antibody (HA721192) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (HA721192) at 1/200 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: Immunocytochemistry analysis of SK-Br-3 cells labeling eIF4E with Rabbit anti-eIF4E antibody (HA721192) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-eIF4E antibody (HA721192) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Fig9: Flow cytometric analysis of SK-Br-3 cells labeling eIF4E.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA721192, 1ug/ml) (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).

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