EEA1 Recombinant Rabbit Monoclonal Antibody [JE59-34]
cat.: HA722147
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat, Monkey
Applications: WB, IF-Cell, FC
Clonality: Monoclonal
Clone number: JE59-34
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: 162 kDa
Isotype: IgG
Immunogen: Recombinant protein within Human EEA1 aa 1-100 / 1,411.
Positive control: HeLa cell lysate, Jurkat cell lysate, JAR cell lysate, NIH/3T3 cell lysate, C6 cell lysate, COS-1 cell lysate, JAR, NIH/3T3, C6.
Subcellular location: Cytoplasm, Early endosome membrane.
Recommended Dilutions:
  WB
  IF-Cell
  FC
1:1,000
1:100
1:1,000
Uniprot #: SwissProt: Q15075 Human | Q8BL66 Mouse
Entrez Gene: 314764 Rat
Alternative names: Early endosome antigen 1 Early endosome antigen 1, 162kD Early endosome associated protein EEA 1 EEA1 EEA1_HUMAN Endosome associated protein p162 Endosome-associated protein p162 MST105 MSTP105 ZFYVE2 Zinc finger FYVE domain containing protein 2 Zinc finger FYVE domain-containing protein 2
Images
HA722147_1.jpg Fig1: Western blot analysis of EEA1 on different lysates with Rabbit anti-EEA1 antibody (HA722147) at 1/1,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: Jurkat cell lysate
Lane 3: JAR cell lysate
Lane 4: NIH/3T3 cell lysate
Lane 5: C6 cell lysate
Lane 6: COS-1 cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 162 kDa
Observed band size: 162 kDa

Exposure time: 12 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 (HA722147) 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.
HA722147_2.jpg Fig2: Western blot analysis of EEA1 on different lysates with Rabbit anti-EEA1 antibody (HA722147) at 1/1,000 dilution.

Lane 1: HAP1-parental cell lysate
Lane 2: HAP1-EEA1 KD cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 162 kDa
Observed band size: 162 kDa

Exposure time: 120 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 (HA722147) at 1/1,000 dilution was used in K1803 at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
HA722147_3.jpg Fig3: Immunocytochemistry analysis of JAR cells labeling EEA1 with Rabbit anti-EEA1 antibody (HA722147) 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-EEA1 antibody (HA722147) 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.
HA722147_4.jpg Fig4: Immunocytochemistry analysis of NIH/3T3 cells labeling EEA1 with Rabbit anti-EEA1 antibody (HA722147) 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-EEA1 antibody (HA722147) 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.
HA722147_5.jpg Fig5: Immunocytochemistry analysis of C6 cells labeling EEA1 with Rabbit anti-EEA1 antibody (HA722147) 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-EEA1 antibody (HA722147) 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.
HA722147_6.jpg Fig6: Flow cytometric analysis of JAR cells labeling EEA1.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA722147, 1μg/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).
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