Anti-UBE1C/UBA3 Antibody Picoband™

Product Name

Anti-UBE1C/UBA3 Antibody Picoband™

View all UBA3 Antibodies

SKU/Catalog Number

PB9838

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-UBE1C/UBA3 Antibody Picoband™ catalog # PB9838. Tested in Flow Cytometry, IF, IHC, ICC, WB applications. This antibody reacts with Human, Mouse, Rat.

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-UBE1C/UBA3 Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # PB9838)

Host

Rabbit

Contents

Each vial contains 5mg BSA, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.

Clonality

Polyclonal

Isotype

Rabbit IgG

Immunogen

A synthetic peptide corresponding to a sequence at the C-terminus of human UBE1C, identical to the related mouse and rat sequences.

*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

PB9838 is reactive to UBA3 in Human, Mouse, Rat

Applications

PB9838 is guaranteed for Flow Cytometry, IF, IHC, ICC, WB Boster Guarantee

Observed Molecular Weight

52 kDa

Calculated molecular weight

51.852kDa

Background of UBA3

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, an ubiquitin-like protein, which regulates cell division, signaling and embryogenesis. Multiple alternatively spliced transcript variants encoding distinct isoforms have been found for this gene.

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.

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Reconsitution

Add 0.2ml of distilled water will yield a concentration of 500ug/ml.

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, 0.1-0.5μg/ml, Human, Rat
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Mouse, Rat, By Heat
Immunocytochemistry/Immunofluorescence, 2μg/ml, Human
Flow Cytometry, 1-3μg/1x10⁶ cells, Human

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of UBE1C using anti-UBE1C antibody (PB9838).
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 Hela whole cell lysate,
Lane 2: human HEK293 whole cell lysate,
Lane 3: rat brain tissue lysate.
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-UBE1C antigen affinity purified polyclonal antibody (Catalog # PB9838) 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 UBE1C at approximately 52 kDa. The expected band size for UBE1C is at 52 kDa.

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Figure 2. IHC analysis of UBE1C using anti-UBE1C antibody (PB9838).
UBE1C was detected in a paraffin-embedded section of rat testis tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 μg/ml rabbit anti-UBE1C Antibody (PB9838) 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 UBE1C using anti-UBE1C antibody (PB9838).
UBE1C was detected in a paraffin-embedded section of mouse testis tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 μg/ml rabbit anti-UBE1C Antibody (PB9838) 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. IF analysis of UBE1C using anti-UBE1C antibody (PB9838).
UBE1C was detected in immunocytochemical section of A431 cell. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2μg/mL rabbit anti-UBE1C Antibody (PB9838) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

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Figure 5. Flow Cytometry analysis of A549 cells using anti-UBE1C antibody (PB9838).
Overlay histogram showing A549 cells stained with PB9838 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-UBE1C Antibody (PB9838,1μg/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 (Red line) was also used as a control.

Gene/Protein Information For UBA3 (Source: Uniprot.org, NCBI)

Alternative Names

DKFZp566J164; EC 6.3.2; EC 6.3.2.-; hUBA3; MGC22384; NEDD8-activating enzyme E1 catalytic subunit; NEDD8-activating enzyme E1C; Nedd8-activating enzyme hUba3; UBA3; UBA3, ubiquitin-activating enzyme E1 homolog; UBE1C; Ubiquitin-activating enzyme 3; ubiquitin-activating enzyme E1C (homologous to yeast UBA3); Ubiquitin-activating enzyme E1C; ubiquitin-like modifier activating enzyme 3; Ubiquitin-like modifier-activating enzyme 3; yeast) UBA3 NAE2, UBE1C, hUBA3 ubiquitin like modifier activating enzyme 3 NEDD8-activating enzyme E1 catalytic subunit|NEDD8-activating enzyme E1 subunit 2|NEDD8-activating enzyme E1C|Nedd8-activating enzyme hUba3|UBA3, ubiquitin-activating enzyme E1 homolog|ubiquitin-activating enzyme 3|ubiquitin-activating enzyme E1C (UBA3 homolog, yeast)|ubiquitin-activating enzyme E1C (homologous to yeast UBA3)

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