UBA3 Mouse Monoclonal Antibody [1B5-2-3]
cat.: EM1901-66
Product Type: Mouse monoclonal IgG2a, primary antibodies
Species reactivity: Human, Mouse
Applications: WB, IF-Cell, IHC-P, FC
Clonality: Monoclonal
Clone number: 1B5-2-3
Form: Liquid
Storage condition: Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.
Storage buffer: 1*PBS (pH7.4), 0.2% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 2ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 52 kDa
Isotype: IgG2a
Immunogen: Recombinant protein within Human UBA3 aa 220-424 / 463.
Positive control: 293T cell lysate, HepG2 cell lysate, EA.hy926, F9, SiHa, human lung tissue, human lung carcinoma tissue, human skin tissue, human breast tissue, human esophagus tissue, SiHa cells.
Subcellular location: Cytosol, nucleus, cytoplasm.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P
  FC
1:5,000
1:50-1:100
1:100-1:500
1:50-1:100
Uniprot #: SwissProt: Q8TBC4 Human
Alternative names: DKFZp566J164 EC 6.3.2. hUba3 MGC22384 NEDD8 activating enzyme E1 catalytic subunit NEDD8 activating enzyme E1C Nedd8 activating enzyme hUba3 NEDD8-activating enzyme E1 catalytic subunit NEDD8-activating enzyme E1C uba3 UBA3 ubiquitin activating enzyme E1 homolog UBA3_HUMAN UBE1C Ubiquitin activating enzyme 3 Ubiquitin activating enzyme E1C Ubiquitin-activating enzyme 3 Ubiquitin-activating enzyme E1C Ubiquitin-like modifier-activating enzyme 3
Images
EM1901-66_1.jpg Fig1: ICC staining of UBA3 in EA.hy926 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (EM1901-66, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Mouse IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
EM1901-66_2.jpg Fig2: ICC staining of UBA3 in F9 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (EM1901-66, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Mouse IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
EM1901-66_3.jpg Fig3: ICC staining of UBA3 in SiHa cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (EM1901-66, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Mouse IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
EM1901-66_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded human lung tissue using anti-UBA3 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM1901-66, 1/200) for 30 minutes 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.
EM1901-66_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded human lung carcinoma tissue using anti-UBA3 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM1901-66, 1/200) for 30 minutes 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.
EM1901-66_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded human skin tissue using anti-UBA3 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM1901-66, 1/200) for 30 minutes 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.
EM1901-66_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded human breast tissue using anti-UBA3 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM1901-66, 1/200) for 30 minutes 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.
EM1901-66_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded human esophagus tissue using anti-UBA3 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM1901-66, 1/200) for 30 minutes 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.
EM1901-66_9.jpg Fig9: Flow cytometric analysis of UBA3 was done on SiHa cells. The cells were fixed, permeabilized and stained with the primary antibody (EM1901-66, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Mouse IgG Secondary antibody at 1/1,000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
EM1901-66_10.jpg Fig10: Western blot analysis of UBA3 on different lysates with Mouse anti-UBA3 antibody (EM1901-66) at 1/5,000 dilution.

Lane 1: HEK-293 (Human mbryonic kidney cell) cell lysate

Exposure time: 120 seconds; ECL: K1801

Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: EM1901-66, 1/5,000 in primary antibody dilution buffer (K1803), overnight at 4 ℃.
Secondary antibody: Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.

Predicted band size: 52 kDa
Observed band size: 52 kDa
EM1901-66_11.jpg Fig11: Western blot analysis of UBA3 on different lysates with Mouse anti-UBA3 antibody (EM1901-66) at 1/5,000 dilution.

Lane 1: HL-60 (Human leukemia cell) cell lysate

Exposure time: 120 seconds; ECL: K1801

Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: EM1901-66, 1/5,000 in primary antibody dilution buffer (K1803), overnight at 4 ℃.
Secondary antibody: Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.

Predicted band size: 52 kDa
Observed band size: 52 kDa
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.