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Product Info Summary
| SKU: | A02295-1 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human, Mouse, Rat |
| Host: | Rabbit |
| Application: | ELISA, Flow Cytometry, IF, IHC, ICC, WB |
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Product info
Product Name
Anti-UBE2I/UBC9 Antibody Picoband®
SKU/Catalog Number
A02295-1
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-UBE2I/UBC9 Antibody Picoband® catalog # A02295-1. Tested in ELISA, Flow Cytometry, IF, IHC, ICC, WB applications. This antibody reacts with Human, Mouse, 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
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-UBE2I/UBC9 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A02295-1)
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 UBE2I/UBC9 recombinant protein (Position: K65-K146 ).
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
A02295-1 is reactive to UBE2I in Human, Mouse, Rat
Observed Molecular Weight
18 kDa
Calculated molecular weight
18.0 kDa
Background of UBE2I
SUMO-conjugating enzyme UBC9 (UBE2I), also called UBC9, is a protein that in humans is encoded by the UBE2I gene. This gene encodes a member of the E2 ubiquitin-conjugating enzyme family. It is mapped to 16p13.3. UBC9 could fully complement the mutant phenotype of a yeast ubc9 mutant strain. This gene may play a similar role via interaction with WT1, which is able to impose a block to cell cycle progression in eukaryotic cells. What’s more, it could support the growth of yeast ubc9 temperature-sensitive mutants at nonpermissive temperatures, indicating that the gene is a functional homolog of yeast ubc9. UBC9 is specifically associated with FHIT, such as FHIT may be involved in cell cycle control through its interaction with UBC9.
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
A02295-1 is guaranteed for ELISA, Flow Cytometry, IF, IHC, ICC, 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, Rat
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Human, Mouse, Rat
Immunocytochemistry/Immunofluorescence, 5μg/ml, Human
Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human
ELISA, 0.1-0.5μg/ml, -
Positive Control
WB: human placenta tissue lysates, human K562 whole cell lysates, human HepG2 whole cell, rat kidney tissue
IHC: human colon cancer tissue, human colon cancer tissue, human mammary cancer tissue, human tonsil tissue, mouse testis tissue, rat testis tissue, rat testis tissue
ICC/IF: A431 cell
FCM: A431 cell
Validation Images & Assay Conditions
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Western blot analysis of UBC9/UBE2I using anti-UBC9/UBE2I antibody (A02295-1).
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 placenta tissue lysates,
Lane 2: human K562 whole cell lysates,
Lane 3: human HepG2 whole cell lysates,
Lane 4: rat kidney 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-UBC9/UBE2I antigen affinity purified polyclonal antibody (A02295-1) 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 UBC9/UBE2I at approximately 18 kDa. The expected band size for UBC9/UBE2I is at 18 kDa.
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IHC analysis of UBE2I/UBC9 using anti-UBE2I/UBC9 antibody (A02295-1).
UBE2I/UBC9 was detected in paraffin-embedded section of human colon 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-UBE2I/UBC9 Antibody (A02295-1) 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.
Click image to see more details
IHC analysis of UBE2I/UBC9 using anti-UBE2I/UBC9 antibody (A02295-1).
UBE2I/UBC9 was detected in paraffin-embedded section of human colon 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-UBE2I/UBC9 Antibody (A02295-1) 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.
Click image to see more details
IHC analysis of UBE2I/UBC9 using anti-UBE2I/UBC9 antibody (A02295-1).
UBE2I/UBC9 was detected in paraffin-embedded section of human mammary 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-UBE2I/UBC9 Antibody (A02295-1) 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.
Click image to see more details
IHC analysis of UBE2I/UBC9 using anti-UBE2I/UBC9 antibody (A02295-1).
UBE2I/UBC9 was detected in paraffin-embedded section of human tonsil 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-UBE2I/UBC9 Antibody (A02295-1) 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.
Click image to see more details
IHC analysis of UBE2I/UBC9 using anti-UBE2I/UBC9 antibody (A02295-1).
UBE2I/UBC9 was detected in paraffin-embedded section of mouse testis 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-UBE2I/UBC9 Antibody (A02295-1) 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.
Click image to see more details
IHC analysis of UBE2I/UBC9 using anti-UBE2I/UBC9 antibody (A02295-1).
UBE2I/UBC9 was detected in paraffin-embedded section of rat testis 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-UBE2I/UBC9 Antibody (A02295-1) 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.
Click image to see more details
IHC analysis of UBE2I/UBC9 using anti-UBE2I/UBC9 antibody (A02295-1).
UBE2I/UBC9 was detected in paraffin-embedded section of rat testis 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-UBE2I/UBC9 Antibody (A02295-1) 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.
Click image to see more details
IF analysis of UBE2I/UBC9 using anti-UBE2I/UBC9 antibody (A02295-1).
UBE2I/UBC9 was detected in immunocytochemical section of A431 cells. 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 5μg/mL rabbit anti-UBE2I/UBC9 Antibody (A02295-1) 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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Flow Cytometry analysis of A431 cells using anti-UBE2I/UBC9 antibody (A02295-1).
Overlay histogram showing A431 cells stained with A02295-1 (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-UBE2I/UBC9 Antibody (A02295-1, 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.
Specific Publications For Anti-UBE2I/UBC9 Antibody Picoband® (A02295-1)
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