UBA3 Mouse Monoclonal Antibody [1C10-5-5]
cat.: EM1901-65
Product Type: Mouse monoclonal IgG1, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IHC-P, FC
Clonality: Monoclonal
Clone number: 1C10-5-5
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
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: IgG1
Immunogen: Recombinant protein within Human UBA3 aa 220-424 / 463.
Positive control: HeLa cell lysate, 293T cell lysate, HepG2 cell lysate, K-562 cell lysate, HL-60 cell lysate, Jurkat cell lysate, mouse brain tissue lysate, rat brain tissue lysate, human brain tissue, mouse brain tissue, rat brain tissue, human lung tissue, human lung carcinoma tissue, human skin tissue, human breast carcinoma tissue, human esophagus tissue, SiHa.
Subcellular location: Cytosol, nucleus, cytoplasm.
Recommended Dilutions:
  WB
  IHC-P
  FC
1:500-1:2,000
1:200-1:2,000
1:50-1:100
Uniprot #: SwissProt: Q8TBC4 Human | Q8C878 Mouse | Q99MI7 Rat
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
EM1901-65_1.jpg Fig1: Western blot analysis of UBA3 on different lysates with Mouse anti-UBA3 antibody (EM1901-65) at 1/1,000 dilution.

Lane 1: HeLa cell lysate (20 µg/Lane)
Lane 2: 293T cell lysate (20 µg/Lane)
Lane 3: HepG2 cell lysate (20 µg/Lane)
Lane 4: K-562 cell lysate (20 µg/Lane)
Lane 5: HL-60 cell lysate (20 µg/Lane)
Lane 6: Jurkat cell lysate (20 µg/Lane)
Lane 7: Mouse brain tissue lysate (40 µg/Lane)
Lane 8: Rat brain tissue lysate (40 µg/Lane)

Predicted band size: 52 kDa
Observed band size: 52 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 (EM1901-65) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.
EM1901-65_2.jpg Fig2: Immunohistochemical analysis of paraffin-embedded human brain tissue with Mouse anti-UBA3 antibody (EM1901-65) at 1/2,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 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 (EM1901-65) at 1/2,000 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.
EM1901-65_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Mouse anti-UBA3 antibody (EM1901-65) at 1/2,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 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 (EM1901-65) at 1/2,000 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.
EM1901-65_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded rat brain tissue with Mouse anti-UBA3 antibody (EM1901-65) at 1/2,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 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 (EM1901-65) at 1/2,000 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.
EM1901-65_5.jpg Fig5: 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) for 20 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 (EM1901-65, 1/400) 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.
EM1901-65_6.jpg Fig6: 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) for 20 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 (EM1901-65, 1/400) 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.
EM1901-65_7.jpg Fig7: 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) for 20 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 (EM1901-65, 1/400) 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.
EM1901-65_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using anti-UBA3 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 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 (EM1901-65, 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.
EM1901-65_9.jpg Fig9: 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) for 20 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 (EM1901-65, 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.
EM1901-65_10.jpg Fig10: Flow cytometric analysis of UBA3 was done on SiHa cells. The cells were fixed, permeabilized and stained with the primary antibody (EM1901-65, 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).
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