UBA3 Mouse Monoclonal Antibody [1B5-2-3]
Catalog# EM1901-66
UBA3 Mouse Monoclonal Antibody [1B5-2-3]
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WB
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IF-Cell
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IHC-P
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FC
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Human
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Mouse
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unconjugated
Overview
Product Name
UBA3 Mouse Monoclonal Antibody [1B5-2-3]
Antibody Type
Mouse Monoclonal Antibody
Immunogen
Recombinant protein within Human UBA3 aa 220-424 / 463.
Species Reactivity
Human, Mouse
Validated Applications
WB, IF-Cell, IHC-P, FC
Target Molecular Weight
Predicted band size: 52 kDa
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.
Conjugation
unconjugated
Clone Number
1B5-2-3
RRID
Product Features
Form
Liquid
Concentration
2 mg/mL.(The concentration of this product may be batch-dependent)
Storage Instructions
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.
Isotype
IgG2a
Purification Method
Protein A affinity purified.
Application Dilution
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WB
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1:5,000
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IF-Cell
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1:50-1:100
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IHC-P
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1:100-1:500
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FC
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1:50-1:100
Target
Function
The modification of proteins with ubiquitin is an important cellular mechanism for targeting abnormal or short-lived proteins for degradation. Ubiquitination involves at least three classes of enzymes: ubiquitin-activating enzymes, or E1s, ubiquitin-conjugating enzymes, or E2s, and ubiquitin-protein ligases, or E3s. This gene encodes a member of the E1 ubiquitin-activating enzyme family. The encoded enzyme associates with AppBp1, an amyloid beta precursor protein binding protein, to form a heterodimer, and then the enzyme complex activates NEDD8, a ubiquitin-like protein, which regulates cell division, signaling and embryogenesis. Multiple alternatively spliced transcript variants encoding distinct isoforms have been found for this gene.
Background References
1. Xu GW. et. al. Mutations in UBA3 confer resistance to the NEDD8-activating enzyme inhibitor MLN4924 in human leukemic cells. PLoS One. 2014 Apr 1;9(4):e93530.
2. Elgin ES. et. al. E2-binding surface on Uba3 β-grasp domain undergoes a conformational transition. Proteins. 2012 Oct;80(10):2482-7.
Sequence Similarity
Belongs to the ubiquitin-activating E1 family. UBA3 subfamily.
Tissue Specificity
Ubiquitously expressed.
Subcellular Location
Cytosol, nucleus, cytoplasm.
UNIPROT
Synonyms
DKFZp566J164 antibody
EC 6.3.2. antibody
hUba3 antibody
MGC22384 antibody
NEDD8 activating enzyme E1 catalytic subunit antibody
NEDD8 activating enzyme E1C antibody
Nedd8 activating enzyme hUba3 antibody
NEDD8-activating enzyme E1 catalytic subunit antibody
NEDD8-activating enzyme E1C antibody
uba3 antibody
ExpandImages
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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).
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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).
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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).
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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.
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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.
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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.
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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.
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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.
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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).
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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 -
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
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
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