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Western Blot (WB) (Figure 1. Western blot analysis of UBE3A using anti-UBE3A antibody (MBS1753973).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 50ug of sample under reducing conditions.Lane 1: human Jurkat whole cell lysatesLane 2: human HEK293 whole cell lysatesLane 3: human Raji whole cell lysatesLane 4: monkey COS-7 whole cell lysates.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1. 5 hour at RT. The membrane was incubated with mouse anti- UBE3A antigen affinity purified monoclonal antibody (Catalog # MBS1753973) at 0. 5 μg/mL overnight at 4 degree C, then washed with TBS-0. 1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # MBS176445) with Tanon 5200 system. A specific band was detected for UBE3A at approximately 100KD. The expected band size for UBE3A is at 100KD. )

Mouse UBE3A Monoclonal Antibody | anti-UBE3A antibody

Anti-UBE3A Antibody (monoclonal, 3F13)

Gene Names
UBE3A; AS; ANCR; E6-AP; HPVE6A; EPVE6AP; FLJ26981; UBE3A
Reactivity
Human, Monkey
Applications
Western Blot, Flow Cytometry, Functional Assay
Purity
Immunogen affinity purified.
Synonyms
UBE3A; Monoclonal Antibody; Anti-UBE3A Antibody (monoclonal; 3F13); E6AP; EPVE6AP; HPVE6A; Ubiquitin-protein ligase E3A; EC 2. 3. 2. 26; 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; ubiquitin protein ligase E3A; anti-UBE3A antibody
Ordering
For Research Use Only!
Host
Mouse
Reactivity
Human, Monkey
Clonality
Monoclonal
Isotype
Mouse IgG2b
Clone Number
3F13
Specificity
Mouse IgG monoclonal antibody for UBE3A detection.
Purity/Purification
Immunogen affinity purified.
Form/Format
Lyophilized. Each vial contains 4mg Trehalose, 0.9mg NaCl and 0.2mg Na2HPO4.
Applicable Applications for anti-UBE3A antibody
Western Blot (WB), Flow Cytometry (FC/FACS/FCM)
Application Notes
WB: 0.25-0.5ug/ml|Human, Monkey|
FC/FACS/FCM: 1-3ug/1x106 cells|Human|
Immunogen
E Coli-derived human UBE3A recombinant protein (Position: M1-E860).
Reconstitution
Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Recommended Detection Systems
Recommended Detection Systems
Preparation and Storage
Store at -20 degree C for one year. After reconstitution, at 4 degree C for one month. It can also be aliquotted and stored frozen at -20 degree C for a longer time. Avoid repeated freezing and thawing.

Western Blot (WB)

(Figure 1. Western blot analysis of UBE3A using anti-UBE3A antibody (MBS1753973).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 50ug of sample under reducing conditions.Lane 1: human Jurkat whole cell lysatesLane 2: human HEK293 whole cell lysatesLane 3: human Raji whole cell lysatesLane 4: monkey COS-7 whole cell lysates.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1. 5 hour at RT. The membrane was incubated with mouse anti- UBE3A antigen affinity purified monoclonal antibody (Catalog # MBS1753973) at 0. 5 μg/mL overnight at 4 degree C, then washed with TBS-0. 1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # MBS176445) with Tanon 5200 system. A specific band was detected for UBE3A at approximately 100KD. The expected band size for UBE3A is at 100KD. )

Western Blot (WB) (Figure 1. Western blot analysis of UBE3A using anti-UBE3A antibody (MBS1753973).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 50ug of sample under reducing conditions.Lane 1: human Jurkat whole cell lysatesLane 2: human HEK293 whole cell lysatesLane 3: human Raji whole cell lysatesLane 4: monkey COS-7 whole cell lysates.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1. 5 hour at RT. The membrane was incubated with mouse anti- UBE3A antigen affinity purified monoclonal antibody (Catalog # MBS1753973) at 0. 5 μg/mL overnight at 4 degree C, then washed with TBS-0. 1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # MBS176445) with Tanon 5200 system. A specific band was detected for UBE3A at approximately 100KD. The expected band size for UBE3A is at 100KD. )

Flow Cytometry (FC/FACS)

(Figure 2. Flow Cytometry analysis of U87 cells using anti-UBE3A antibody (MBS1753973).Overlay histogram showing U87 cells stained with MBS1753973 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti- UBE3A Antibody (MBS1753973, 1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was mouse IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control. )

Flow Cytometry (FC/FACS) (Figure 2. Flow Cytometry analysis of U87 cells using anti-UBE3A antibody (MBS1753973).Overlay histogram showing U87 cells stained with MBS1753973 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti- UBE3A Antibody (MBS1753973, 1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was mouse IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control. )
Related Product Information for anti-UBE3A antibody
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.
References
1. Abaied, L., Trabelsi, M., Chaabouni, M., Kharrat, M., Kraoua, L., M'rad, R., Tebib, N., Maazoul, F., Chaabouni, H. A novel UBE3A truncating mutation in large Tunisian Angelman syndrome pedigree. Am. J. Med. Genet. 152A: 141-146, 2010.
2. Albrecht, U., Sutcliffe, J. S., Cattanach, B. M., Beechey, C. V., Armstrong, D., Eichele, G., Beaudet, A. L. Imprinted expression of the murine Angelman syndrome gene, Ube3a, in hippocampal and Purkinje neurons. Nature Genet. 17: 75-78, 1997.
3. Camprubi, C., Guitart, M., Gabau, E., Coll, M. D., Villatoro, S., Oltra, S., Rosello, M., Ferrer, I., Monfort, S., Orellana, C., Martinez, F. Novel UBE3A mutations causing Angelman syndrome: different parental origin for single nucleotide changes and multiple nucleotide deletions or insertions. Am. J. Med. Genet. 149A: 343-348, 2009.

NCBI and Uniprot Product Information

NCBI GI #
NCBI GeneID
UniProt Accession #
NCBI Official Full Name
ubiquitin-protein ligase E3A isoform 2 [Homo sapiens]
NCBI Official Synonym Full Names
ubiquitin protein ligase E3A
NCBI Official Symbol
UBE3A
NCBI Official Synonym Symbols
AS; ANCR; E6-AP; HPVE6A; EPVE6AP; FLJ26981; UBE3A
UniProt Protein Name
Ubiquitin-protein ligase E3A
Protein Family
UniProt Gene Name
UBE3A
UniProt Synonym Gene Names
E6AP; EPVE6AP; HPVE6A
UniProt Entry Name
UBE3A_HUMAN

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Product Notes

The UBE3A ube3a (Catalog #AAA1753973) is an Antibody produced from Mouse and is intended for research purposes only. The product is available for immediate purchase. The Anti-UBE3A Antibody (monoclonal, 3F13) reacts with Human, Monkey and may cross-react with other species as described in the data sheet. AAA Biotech's UBE3A can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB), Flow Cytometry (FC/FACS/FCM). WB: 0.25-0.5ug/ml|Human, Monkey| FC/FACS/FCM: 1-3ug/1x106 cells|Human|. Researchers should empirically determine the suitability of the UBE3A ube3a for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "UBE3A, Monoclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.

Precautions

All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.

Disclaimer

Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.

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