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Product Info Summary
| SKU: | A03118-2 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human, Rat |
| Host: | Rabbit |
| Application: | ELISA, Flow Cytometry, WB |
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Product info
Product Name
Anti-BUB3 Antibody Picoband®
SKU/Catalog Number
A03118-2
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-BUB3 Antibody Picoband® catalog # A03118-2. Tested in ELISA, Flow Cytometry, WB applications. This antibody reacts with Human, Rat. 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
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 freezing and thawing.
Cite This Product
Anti-BUB3 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A03118-2)
Host
Rabbit
Contents
Each vial contains 4 mg Trehalose, 0.9 mg NaCl, 0.2 mg Na2HPO4.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E.coli-derived human BUB3 recombinant protein (Position: R50-D209).
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
A03118-2 is reactive to BUB3 in Human, Rat
Observed Molecular Weight
40 kDa
Calculated molecular weight
37.2 kDa
Background of BUB3
BUB3(BUDDING UNINHIBITED BY BENZIMIDAZOLES 3) is a protein that in humans is encoded by the BUB3 gene. And it is mapped on 10q26.13. Bub3 is also a protein involved with the regulation of the Spindle Assembly Checkpoint(SAC); though BUB3 is non-essential in yeast, it is essential in higher eukaryotes. As one of the checkpoint proteins, Bub3 delays the irreversible onset of anaphase through direction of kinetochore localization during prometaphase to achieve biorentation. Bub3 directs the localization of Bub1 at the kinetochore in order to activate the SAC. Bub3 also acts as a regulator in that it affects binding of Mad3 to Mad2. Using deletion analysis, the authors identified a domain of BUB1 that is required both for binding BUB3 and for kinetochore localization of BUB1.
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
A03118-2 is guaranteed for ELISA, Flow Cytometry, 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, 0.25-0.5 μg/ml, Human, Rat
Flow Cytometry (Fixed), 1-3 μg/1x106 cells, Human
ELISA, 0.1-0.5 μg/ml, -
Positive Control
WB: human Hela whole cell, human 293T whole cell, rat thymus tissue
FCM: A431 cell
Validation Images & Assay Conditions
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Western blot analysis of BUB3 using anti-BUB3 antibody (A03118-2).
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 lysates,
Lane 2: human 293T whole cell lysates,
Lane 3: rat thymus tissue 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-BUB3 antigen affinity purified polyclonal antibody (Catalog # A03118-2) 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 BUB3 at approximately 40 kDa. The expected band size for BUB3 is at 37 kDa.
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Flow Cytometry analysis of A431 cells using anti-BUB3 antibody (A03118-2).
Overlay histogram showing A431 cells stained with A03118-2 (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-BUB3 Antibody (A03118-2, 1 μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
Specific Publications For Anti-BUB3 Antibody Picoband® (A03118-2)
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