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Product Info Summary
| SKU: | A00582 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human |
| Host: | Rabbit |
| Application: | ELISA, Flow Cytometry, IHC, WB |
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Product info
Product Name
Anti-UBE3A Picoband® Antibody
View all E6AP/UBE3A Antibodies
SKU/Catalog Number
A00582
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-UBE3A Picoband® Antibody catalog # A00582. Tested in ELISA, Flow Cytometry, IHC, WB applications. This antibody reacts with Human. The brand Picoband indicates this is a premium antibody that guarantees superior quality, high affinity, and strong signals with minimal background in Western blot applications. Only our best-performing antibodies are designated as Picoband, ensuring unmatched performance.
Storage & Handling
Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.
Cite This Product
Anti-UBE3A Picoband® Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # A00582)
Host
Rabbit
Contents
Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E.coli-derived human UBE3A recombinant protein (Position: M1-E860).
*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
A00582 is reactive to UBE3A in Human
Reconstitution
Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Observed Molecular Weight
100 kDa
Calculated molecular weight
100 kDa
Background of E6AP/UBE3A
Ubiquitin-protein ligase E3A (UBE3A) also known as E6AP ubiquitin-protein ligase (E6AP) is an enzyme that in humans is encoded by the UBE3A gene. It is mapped to 15q11.2. This gene encodes an E3 ubiquitin-protein ligase, part of the ubiquitin protein degradation system. This imprinted gene is maternally expressed in brain and biallelically expressed in other tissues. Maternally inherited deletion of this gene causes Angelman Syndrome, characterized by severe motor and intellectual retardation, ataxia, hypotonia, epilepsy, absence of speech, and characteristic facies. The protein also interacts with the E6 protein of human papillomavirus types 16 and 18, resulting in ubiquitination and proteolysis of tumor protein p53. Alternative splicing of this gene results in three transcript variants encoding three isoforms with different N-termini. Additional transcript variants have been described, but their full length nature has not been determined.
Antibody Validation
Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.
Application & Images
Applications
A00582 is guaranteed for ELISA, Flow Cytometry, IHC, WB Boster Guarantee
Assay Dilutions Recommendation
The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.
Western blot, 0.25-0.5μg/ml, Human
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Human, Mouse, Rat
Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human
ELISA, 0.1-0.5μg/ml, -
Positive Control
WB: human 293T whole cell, human Jurkat whole cell, human K562 whole cell, human HepG2 whole cell, human Hela whole cell
IHC: human gastric cancer tissue, human rectal cancer tissue
FCM: CACO-2 cell
Validation Images & Assay Conditions
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Figure 1. Western blot analysis of UBE3A using anti-UBE3A antibody (A00582).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human 293T whole cell lysates,
Lane 2: human Jurkat whole cell lysates,
Lane 3: human K562 whole cell lysates,
Lane 4: human HepG2 whole cell lysates,
Lane 5: human Hela whole cell lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-UBE3A antigen affinity purified polyclonal antibody (Catalog # A00582) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for UBE3A at approximately 100 kDa. The expected band size for UBE3A is at 100 kDa.
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Figure 2. IHC analysis of UBE3A using anti-UBE3A antibody (A00582).
UBE3A was detected in paraffin-embedded section of human gastric cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-UBE3A Antibody (A00582) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.
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Figure 3. IHC analysis of UBE3A using anti-UBE3A antibody (A00582).
UBE3A was detected in paraffin-embedded section of human rectal cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-UBE3A Antibody (A00582) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.
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Figure 4. Flow Cytometry analysis of CACO-2 cells using anti-UBE3A antibody (A00582).
Overlay histogram showing CACO-2 cells stained with A00582 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-UBE3A Antibody (A00582, 1μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
Protein Target Info & Infographic
Gene/Protein Information For UBE3A (Source: Uniprot.org, NCBI)
Gene Name
UBE3A
Full Name
Ubiquitin-protein ligase E3A
Weight
100 kDa
Alternative Names
Ubiquitin-protein ligase E3A; E6AP ubiquitin-protein ligase; HECT-type ubiquitin transferase E3A; Human papillomavirus E6-associated protein; Oncogenic protein-associated protein E6-AP; Renal carcinoma antigen NY-REN-54; UBE3A; E6AP; EPVE6AP; HPVE6A UBE3A ANCR, AS, E6-AP, EPVE6AP, HPVE6A ubiquitin protein ligase E3A ubiquitin-protein ligase E3A|CTCL tumor se37-2|E6AP ubiquitin-protein ligase|HECT-type ubiquitin transferase E3A|human papilloma virus E6-associated protein|human papillomavirus E6-associated protein|oncogenic protein-associated protein E6-AP|renal carcinoma NY-REN-54
*If product is indicated to react with multiple species, protein info is based on the gene entry specified above in "Species".For more info on UBE3A, check out the UBE3A Infographic
We have 30,000+ of these available, one for each gene! Check them out.
In this infographic, you will see the following information for UBE3A: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].
Specific Publications For Anti-UBE3A Picoband® Antibody (A00582)
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