eIF4A1 Recombinant Rabbit Monoclonal Antibody [JE35-43]
cat.: HA721430
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IHC-P, IF-Cell, FC
Clonality: Monoclonal
Clone number: JE35-43
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: 46 kDa
Isotype: IgG
Immunogen: Synthetic peptide within Human eIF4A1 aa 1-50 / 406.
Positive control: HeLa cell lysate, U-2 OS cell lysate, K-562 cell lysate, HEK-293 cell lysate, MCF7 cell lysate, HepG2 cell lysate, Huh7 cell lysate, Jurkat cell lysate, PC-12 cell lysate, mouse colon tissue, HeLa.
Subcellular location: Cytoplasm, cytosol, eukaryotic translation initiation factor 4F complex, extracellular exosome, membrane.
Recommended Dilutions:
  WB
  IHC-P
  IF-Cell
  FC

1:1,000
1:200
1:100
1:500-1:1,000
Uniprot #: SwissProt: P60842 Human | P60843 Mouse
Entrez Gene: 287436 Rat
Alternative names: ATP-dependent RNA helicase eIF4A-1 DDX2 DDX2A eIF 4A I eIF-4A-I EIF4A eIF4A I eIF4A-I EIF4A1 Eukaryotic initiation factor 4A-I Eukaryotic initiation factor 4AI eukaryotic translation initiation factor 4A, isoform 1 Eukaryotic translation initiation factor 4A1 IF4A1_HUMAN
Images
HA721430_1.jpg Fig1: Western blot analysis of eIF4A1 on different lysates with Rabbit anti-eIF4A1 antibody (HA721430) at 1/1,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: U-2 OS cell lysate
Lane 3: K-562 cell lysate
Lane 4: HEK-293 cell lysate
Lane 5: MCF7 cell lysate
Lane 6: HepG2 cell lysate
Lane 7: Huh7 cell lysate
Lane 8: Jurkat cell lysate
Lane 9: PC-12 cell lysate

Lysates/proteins at 15 µg/Lane.

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

Exposure time: 3 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 (HA721430) 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 1:100,000 dilution was used for 1 hour at room temperature.
HA721430_2.jpg Fig2: Immunohistochemical analysis of paraffin-embedded mouse colon tissue with Rabbit anti-eIF4A1 antibody (HA721430) 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 (HA721430) 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.
HA721430_3.jpg Fig3: Immunocytochemistry analysis of HeLa cells labeling eIF4A1 with Rabbit anti-eIF4A1 antibody (HA721430) at 1/100 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-eIF4A1 antibody (HA721430) at 1/100 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. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (M1305-2, red) was stained at 1/200 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
HA721430_4.jpg Fig4: Flow cytometric analysis of HeLa cells labeling eIF4A1.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA721430, 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”.