UBA3 Recombinant Mouse Monoclonal Antibody [1B5-2-3-R] - BSA and Azide free
Catalog# HA610041
UBA3 Recombinant Mouse Monoclonal Antibody [1B5-2-3-R] - BSA and Azide free
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WB
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IHC-P
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IF-Cell
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Human
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Mouse
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Rat
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HA601130
含抗保成分
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unconjugated
Overview
Product Name
UBA3 Recombinant Mouse Monoclonal Antibody [1B5-2-3-R] - BSA and Azide free
Antibody Type
Recombinant Mouse Monoclonal Antibody
Immunogen
Recombinant protein within Human UBA3 aa 220-424 / 463.
Species Reactivity
Human, Mouse, Rat
Validated Applications
WB, IHC-P, IF-Cell
Molecular Weight
Predicted band size: 52 kDa
Positive Control
HepG2 cell lysate, Hela cell lysate, 293T cell lysate, K-562 cell lysate, RAW264.7 cell lysate, NIH/3T3 cell lysate, PC-12 cell lysate, mouse brain tissue lysate, human lung tissue lysate, human breast tissue, human esophagus tissue, human lung carcinoma tissue, human lung tissue, human skin tissue, SiHa.
Conjugation
unconjugated
Clone Number
1B5-2-3-R
Product Features
Form
Liquid
Concentration
1mg/ml
Storage Instructions
Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage Buffer
1*PBS (pH7.4).
Isotype
IgG1
Purification Method
Protein A affinity purified.
Application Dilution
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WB
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1:1,000
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IHC-P
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1:1,000
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IF-Cell
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1:200
Target
Function
NEDD8-activating enzyme E1 catalytic subunit is a protein that in humans is encoded by the UBA3 gene. 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. This enzyme contains an E2 binding domain, which resembles ubiquitin, and recruits the catalytic core of the E2 enzyme UBE2M (Ubc12) in a similar manner to that in which ubiquitin interacts with ubiquitin binding domains.
Background References
1. Osaka F, Kawasaki H, Aida N, Saeki M, Chiba T, Kawashima S, Tanaka K, Kato S (August 1998). "A new NEDD8-ligating system for cullin-4A". Genes Dev. 12 (15): 2263–8.
2. Huang DT, Paydar A, Zhuang M, Waddell MB, Holton JM, Schulman BA (February 2005). "Structural basis for recruitment of Ubc12 by an E2 binding domain in NEDD8\'s E1". Mol. Cell. 17 (3): 341–50.
Subcellular Location
Cytosol, nucleus, cytoplasm.
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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Western blot analysis of UBA3 on different lysates with Mouse anti-UBA3 antibody (HA610041) at 1/500 dilution.
Lane 1: HepG2 cell lysate
Lane 2: Hela cell lysate
Lane 3: 293T cell lysate
Lane 4: K-562 cell lysate
Lane 5: RAW264.7 cell lysate
Lane 6: NIH/3T3 cell lysate
Lane 7: PC-12 cell lysate
Lane 8: Mouse brain tissue lysate
Lane 9: Human lung tissue lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 52 kDa
Observed band size: 52 kDa
Exposure time: 2 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 (HA610041) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:150,000 dilution was used for 1 hour at room temperature. -
Immunohistochemical analysis of paraffin-embedded human breast tissue with Mouse anti-UBA3 antibody (HA610041) at 1/1,000 dilution.
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 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA610041) at 1/1,000 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. -
Immunohistochemical analysis of paraffin-embedded human esophagus tissue with Mouse anti-UBA3 antibody (HA610041) at 1/1,000 dilution.
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 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA610041) at 1/1,000 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. -
Immunohistochemical analysis of paraffin-embedded human lung carcinoma tissue with Mouse anti-UBA3 antibody (HA610041) at 1/1,000 dilution.
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 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA610041) at 1/1,000 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. -
Immunohistochemical analysis of paraffin-embedded human lung tissue with Mouse anti-UBA3 antibody (HA610041) at 1/1,000 dilution.
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 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA610041) at 1/1,000 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. -
Immunohistochemical analysis of paraffin-embedded human skin tissue with Mouse anti-UBA3 antibody (HA610041) at 1/1,000 dilution.
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 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA610041) at 1/1,000 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. -
Immunocytochemistry analysis of SiHa cells labeling UBA3 with Mouse anti-UBA3 antibody (HA610041) at 1/200 dilution.
Cells were fixed in 4% paraformaldehyde for 30 minutes, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Mouse anti-UBA3 antibody (HA610041) at 1/200 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.
beta Tubulin (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) were used as the secondary antibody at 1/1,000 dilution.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"