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Product Info Summary
| SKU: | A04728-1 |
|---|---|
| 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-UBE2D1/2/3/4 Antibody Picoband®
View all UbcH5a/UBE2D1 Antibodies
SKU/Catalog Number
A04728-1
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-UBE2D1/2/3/4 Antibody Picoband® catalog # A04728-1. 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-UBE2D1/2/3/4 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A04728-1)
Host
Rabbit
Contents
Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.005mg NaN3.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E.coli-derived human UBE2D1/2/3/4 recombinant protein (Position: D116-M147).
*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
A04728-1 is reactive to UBE2D1/2/3/4 in Human, Mouse, Rat
Reconstitution
Add 0.2ml of distilled water will yield a concentration of 500μg/ml.
Observed Molecular Weight
17 kDa
Calculated molecular weight
32193 MW
Background of UbcH5a/UBE2D1
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 E2 ubiquitin-conjugating enzyme family. This enzyme is closely related to a stimulator of iron transport (SFT), and is up-regulated in hereditary hemochromatosis. It also functions in the ubiquitination of the tumor-suppressor protein p53 and the hypoxia-inducible transcription factor HIF1alpha by interacting with the E1 ubiquitin-activating enzyme and the E3 ubiquitin-protein ligases. Two transcript variants encoding different 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.
Application & Images
Applications
A04728-1 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), 2-5μg/ml, Human, Mouse, Rat
Immunocytochemistry/Immunofluorescence, 5μg/ml, Human
Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human, Mouse, Rat
ELISA, 0.1-0.5μg/ml, -
Positive Control
WB: rat heart tissue, rat lung tissue, rat liver tissue, human HEK293 whole cell, human Jurkat whole cell, human CACO-2 whole cell
IHC: mouse lung lymph node tissue, human testicular cancer tissue, human bladder cancer tissue, rat lung tissue
ICC/IF: A431 cell
FCM: A431 cell, RAW2647 cell, RH35 cell
Validation Images & Assay Conditions
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Figure 1. Western blot analysis of UBE2D1/2/3/4 using anti-UBE2D1/2/3/4 antibody (A04728-1).
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 50ug of sample under reducing conditions.
Lane 1: rat heart tissue lysates,
Lane 2: rat lung tissue lysates,
Lane 3: rat liver tissue lysates,
Lane 4: human HEK293 whole cell lysates,
Lane 5: human Jurkat whole cell lysates,
Lane 6: human CACO-2 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 rabbit anti-UBE2D1/2/3/4 antigen affinity purified polyclonal antibody (Catalog # A04728-1) 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 UBE2D1/2/3/4 at approximately 17KD. The expected band size for UBE2D1/2/3/4 is at 17KD.
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Figure 2. IHC analysis of UBE2D1/2/3/4 using anti-UBE2D1/2/3/4 antibody (A04728-1).
UBE2D1/2/3/4 was detected in paraffin-embedded section of mouse lung lymph node 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 2μg/ml rabbit anti-UBE2D1/2/3/4 Antibody (A04728-1) 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.
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Figure 3. IHC analysis of UBE2D1/2/3/4 using anti-UBE2D1/2/3/4 antibody (A04728-1).
UBE2D1/2/3/4 was detected in paraffin-embedded section of human testicular 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 2μg/ml rabbit anti-UBE2D1/2/3/4 Antibody (A04728-1) 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
Figure 4. IHC analysis of UBE2D1/2/3/4 using anti-UBE2D1/2/3/4 antibody (A04728-1).
UBE2D1/2/3/4 was detected in paraffin-embedded section of human bladder 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 2μg/ml rabbit anti-UBE2D1/2/3/4 Antibody (A04728-1) 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
Figure 5. IHC analysis of UBE2D1/2/3/4 using anti-UBE2D1/2/3/4 antibody (A04728-1).
UBE2D1/2/3/4 was detected in paraffin-embedded section of rat lung 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 2μg/ml rabbit anti-UBE2D1/2/3/4 Antibody (A04728-1) 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
Figure 6. IF analysis of UBE2D1/2/3/4 using anti-UBE2D1/2/3/4 antibody (A04728-1).
UBE2D1/2/3/4 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-UBE2D1/2/3/4 Antibody (A04728-1) 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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Figure 7. Flow Cytometry analysis of A431 cells using anti-UBE2D1/2/3/4 antibody (A04728-1).
Overlay histogram showing A431 cells stained with A04728-1 (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-UBE2D1/2/3/4 Antibody (A04728-1, 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.
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Figure 8. Flow Cytometry analysis of RAW264.7 cells using anti-UBE2D1/2/3/4 antibody (A04728-1).
Overlay histogram showing RAW264.7 cells stained with A04728-1 (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-UBE2D1/2/3/4 Antibody (A04728-1, 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.
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Figure 9. Flow Cytometry analysis of RH35 cells using anti-UBE2D1/2/3/4 antibody (A04728-1).
Overlay histogram showing RH35 cells stained with A04728-1 (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-UBE2D1/2/3/4 Antibody (A04728-1, 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.
Protein Target Info & Infographic
Gene/Protein Information For UBE2D1/2/3/4 (Source: Uniprot.org, NCBI)
Gene Name
UBE2D1/2/3/4
Full Name
Ubiquitin-conjugating enzyme E2 D1
Weight
32193 MW
Superfamily
ubiquitin-conjugating enzyme family
Alternative Names
Homeobox protein Hox-B1;Homeobox protein Hox-2I;HOXB1;HOX2I; UBE2D1 E2(17)KB1, SFT, UBC4/5, UBCH5, UBCH5A ubiquitin conjugating enzyme E2 D1 ubiquitin-conjugating enzyme E2 D1|(E3-independent) E2 ubiquitin-conjugating enzyme D1|E2 ubiquitin-conjugating enzyme D1|UBC4/5 homolog|stimulator of Fe transport|ubiquitin carrier protein D1|ubiquitin conjugating enzyme E2D 1|ubiquitin-conjugating enzyme E2(17)KB 1|ubiquitin-conjugating enzyme E2-17 kDa 1|ubiquitin-conjugating enzyme E2D 1 (UBC4/5 homolog, yeast)|ubiquitin-protein ligase D1
*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 UBE2D1/2/3/4, check out the UBE2D1/2/3/4 Infographic
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In this infographic, you will see the following information for UBE2D1/2/3/4: 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-UBE2D1/2/3/4 Antibody Picoband® (A04728-1)
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