HA721746

eIF4A2 Recombinant Rabbit Monoclonal Antibody [PSH01-88]

cat.: HA721746

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat, Monkey
Applications:	WB, IHC-P, IF-Cell, FC
Clonality:	Monoclonal
Clone number:	PSH01-88

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	1ug/ul
Purification:	Protein A affinity purified.
Molecular weight:	Predicted band size: 46 kDa
Isotype:	IgG
Immunogen:	Recombinant protein within Human eIF4A2 aa1-407.
Positive control:	PC-3M cell lysate, C2C12 cell lysate, COS-1 cell lysate, human brain tissue lysate, mouse testis tissue lysate, PC-12, human testis tissue, mouse brain tissue, mouse testis tissue, rat brain tissue, rat testis tissue, HeLa, NIH/3T3.
Subcellular location:	Cytosol.
Recommended Dilutions: WB IHC-P IF-Cell FC	1:1,000 1:1,000 1:100 1:1,000
Uniprot #:	SwissProt: Q14240 Human \| Q5RKI1 Rat \| P10630 Mouse
Alternative names:	ATP dependent RNA helicase eIF4A 2 ATP-dependent RNA helicase eIF4A-2 BM-010 DDX2B eIF 4A II eIF-4A-II EIF4A eIF4A II eIF4A-II EIF4A2 EIF4F Eukaryotic initiation factor 4A II Eukaryotic initiation factor 4A-II eukaryotic translation initiation factor 4A isoform 2 Eukaryotic translation initiation factor 4A2 IF4A2_HUMAN N-terminally processed

Images

Fig1: Western blot analysis of eIF4A2 on different lysates with Rabbit anti-eIF4A2 antibody (HA721746) at 1/1,000 dilution.

Lane 1: PC-3M cell lysate
Lane 2: C2C12 cell lysate
Lane 3: COS-1 cell lysate
Lane 4: Human brain tissue lysate
Lane 5: Mouse testis tissue lysate

Lysates/proteins at 30 µg/Lane.

Predicted band size: 46.4 kDa
Observed band size: 46 kDa

Exposure time: 1 minutes;

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 (HA721746) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

Fig2: Immunocytochemistry analysis of PC-12 cells labeling eIF4A2 with Rabbit anti-eIF4A2 antibody (HA721746) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-eIF4A2 antibody (HA721746) at 1/100 dilution in 1% BSA in PBST 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.

Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

	Fig3: Immunohistochemical analysis of paraffin-embedded human testis tissue with Rabbit anti-eIF4A2 antibody (HA721746) at 1/1,000 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 (HA721746) 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.
	Fig4: Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Rabbit anti-eIF4A2 antibody (HA721746) at 1/1,000 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 (HA721746) 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.
	Fig5: Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-eIF4A2 antibody (HA721746) at 1/1,000 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 (HA721746) 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.

	Fig6: Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-eIF4A2 antibody (HA721746) at 1/1,000 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 (HA721746) 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 rat testis tissue with Rabbit anti-eIF4A2 antibody (HA721746) at 1/1,000 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 (HA721746) 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.
	Fig8: Flow cytometric analysis of HeLa cells labeling eIF4A2. Cells were fixed and permeabilized. Then stained with the primary antibody (HA721746, 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).
	Fig9: Flow cytometric analysis of PC-12 cells labeling eIF4A2. Cells were fixed and permeabilized. Then stained with the primary antibody (HA721746, 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 eIF4A2.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA721746, 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).

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