HUWE1 Rabbit Polyclonal Antibody
cat.: ER61660
Product Type: Rabbit polyclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, FC
Clonality: Polyclonal
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: 2ug/ul
Purification: Immunogen affinity purified.
Molecular weight: Predicted band size: 482 kDa
Isotype: IgG
Immunogen: Recombinant protein within human HUWE1 aa 3,950-4,374.
Positive control: HeLa cell lysate, 293T cell lysate, A549 cell lysate, NIH/3T3 cell lysate, PC-12 cell lysate, Mouse brain tissue lysate, Rat brain tissue lysate, HeLa, NIH/3T3, PC-12.
Subcellular location: Cytoplasm, Nucleus, Mitochondrion.
Recommended Dilutions:
  WB
  IF-Cell
  FC
1:10,000
1:100-1:200
1:1,000
Uniprot #: SwissProt: Q7Z6Z7 Human | Q7TMY8 Mouse | P51593 Rat
Alternative names: ARF binding protein 1 ARF BP1 ARF-binding protein 1 ARF-BP1 ARFBP1 BJ-HCC-24 tumor antigen E3 ubiquitin protein ligase HUWE1 E3 ubiquitin-protein ligase HUWE1 HECT HECT domain protein LASU1 HECT UBA and WWE domain containing protein 1 HectH9 Homologous to E6AP carboxyl terminus homologous protein 9 Huwe1 HUWE1_HUMAN Ib772 Large structure of UREB1 LASU1 Mcl 1 ubiquitin ligase E3 Mcl-1 ubiquitin ligase E3 Mule UBA and WWE domain-containing protein 1 Upstream regulatory element-binding protein 1 URE-B1 URE-binding protein 1 UREB1
Images
ER61660_1.jpg Fig1: Western blot analysis of HUWE1 on different lysates with Rabbit anti-HUWE1 antibody (ER61660) at 1/10,000 dilution.

Lane 1: HeLa cell lysate (20 µg/Lane)
Lane 2: 293T cell lysate (20 µg/Lane)
Lane 3: A549 cell lysate (low expression) (20 µg/Lane)
Lane 4: NIH/3T3 cell lysate (20 µg/Lane)
Lane 5: PC-12 cell lysate (20 µg/Lane)
Lane 6: Mouse brain tissue lysate (40 µg/Lane)
Lane 7: Rat brain tissue lysate (40 µg/Lane)

Predicted band size: 482 kDa
Observed band size: 482 kDa

Exposure time: 25 seconds; ECL: K1801;

3-8% 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 (ER61660) at 1/10,000 dilution was used in primary antibody dilution (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.
ER61660_2.jpg Fig2: Immunocytochemistry analysis of HeLa cells labeling HUWE1 with Rabbit anti-HUWE1 antibody (ER61660) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-HUWE1 antibody (ER61660) 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 (HA601187, 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.
ER61660_3.jpg Fig3: Immunocytochemistry analysis of NIH/3T3 cells labeling HUWE1 with Rabbit anti-HUWE1 antibody (ER61660) at 1/200 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-HUWE1 antibody (ER61660) at 1/200 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 (HA601187, 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.
ER61660_4.jpg Fig4: Immunocytochemistry analysis of PC-12 cells labeling HUWE1 with Rabbit anti-HUWE1 antibody (ER61660) at 1/200 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-HUWE1 antibody (ER61660) at 1/200 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 (HA601187, 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.
ER61660_5.jpg Fig5: Flow cytometric analysis of PC-12 cells labeling HUWE1.

Cells were fixed and permeabilized. Then stained with the primary antibody (ER61660, 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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