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Product Info Summary
| SKU: | A00547 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human, Mouse, Rat |
| Host: | Rabbit |
| Application: | Flow Cytometry, IF, IHC, ICC, WB |
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Product info
Product Name
Anti-NEDD8 Antibody Picoband®
SKU/Catalog Number
A00547
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-NEDD8 Antibody Picoband® catalog # A00547. Tested in 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-NEDD8 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A00547)
Host
Rabbit
Contents
Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
A synthetic peptide corresponding to a sequence in the middle region of human NEDD8, 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
A00547 is reactive to NEDD8 in Human, Mouse, Rat
Reconstitution
Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Observed Molecular Weight
12 kDa
Calculated molecular weight
9072 MW
Background of NEDD8
NEDD8 is a protein that in humans is encoded by the NEDD8 gene. Human NEDD8 shares 60% amino acid sequence identity to ubiquitin. The most substrates of NEDD8 modification are the Cullin subunits of Cullin-based E3 ubiquitin ligases, which are active only when neddylated. Their NEDDylation is critical for the recruitment of E2 to the ligase complex, thus facilitating ubiquitin conjugation. NEDD8 modification has therefore been implicated in cell cycle progression and cytoskeletal regulation.
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
A00547 is guaranteed for 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, 0.1-0.5μg/ml, Human, Mouse, Rat
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Human, By Heat
Immunocytochemistry/Immunofluorescence, 2μg/ml, Human
Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human
Positive Control
WB: human HepG2 whole cell, human MCF-7 whole cell, human RT4 whole cell, human PC-3 whole cell, rat brain tissue, rat thymus tissue, mouse brain tissue, mouse thymus tissue
IHC: human lung cancer tissue
ICC/IF: A431 cell
FCM: A431 cell
Validation Images & Assay Conditions
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Figure 1. Western blot analysis of NEDD8 using anti-NEDD8 antibody (A00547).
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 HepG2 whole cell lysates,
Lane 2: human MCF-7 whole cell lysates,
Lane 3: human RT4 whole cell lysates,
Lane 4: human PC-3 whole cell lysates,
Lane 5: rat brain tissue lysates,
Lane 6: rat thymus tissue lysates,
Lane 7: mouse brain tissue lysates,
Lane 8: mouse 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-NEDD8 antigen affinity purified polyclonal antibody (Catalog # A00547) 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 NEDD8 at approximately 10-12 kDa. The expected band size for NEDD8 is at 9 kDa.
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Figure 2. IHC analysis of NEDD8 using anti-NEDD8 antibody (A00547).
NEDD8 was detected in paraffin-embedded section of human lung cancer tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-NEDD8 Antibody (A00547) 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. Flow Cytometry analysis of A431 cells using anti-NEDD8 antibody (A00547).
Overlay histogram showing A431 cells stained with A00547 (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-NEDD8 Antibody (A00547,1μg/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 4. IF analysis of NEDD8 using anti-NEDD8 antibody (A00547).
NEDD8 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 2μg/mL rabbit anti-NEDD8 Antibody (A00547) 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.
Protein Target Info & Infographic
Gene/Protein Information For NEDD8 (Source: Uniprot.org, NCBI)
Gene Name
NEDD8
Full Name
NEDD8
Weight
9072 MW
Superfamily
ubiquitin family
Alternative Names
NEDD8;Neddylin;Neural precursor cell expressed developmentally down-regulated protein 8;NEDD-8;Ubiquitin-like protein Nedd8;NEDD8; NEDD8 NEDD-8 NEDD8 ubiquitin like modifier NEDD8|neddylin|neural precursor cell expressed, developmentally down-regulated 8|ubiquitin-like protein Nedd8
*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 NEDD8, check out the NEDD8 Infographic
We have 30,000+ of these available, one for each gene! Check them out.
In this infographic, you will see the following information for NEDD8: 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-NEDD8 Antibody Picoband® (A00547)
Hello CJ!
A00547 has been cited in 1 publications:
*The publications in this section are manually curated by our staff scientists. They may differ from Bioz's machine gathered results. Both are accurate. If you find a publication citing this product but is missing from this list, please let us know we will issue you a thank-you coupon.
Jiang B,Adams Z,Moonah S,Shi H,Maupin-Furlow J,Moskovitz J.The Antioxidant Enzyme Methionine Sulfoxide Reductase A (MsrA) Interacts with Jab1/CSN5 and Regulates Its Function.Antioxidants (Basel).2020 May 24;9(5):452.doi:10.3390/antiox9050452.PMID:32456285
Species: Mouse
A00547 usage in article: APP:WB, SAMPLE:BRAIN AND LIVER EXTRACTS, DILUTION:NA
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Customer Q&As
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1 Customer Q&As for Anti-NEDD8 Antibody Picoband®
Question
We are currently using anti-NEDD8 antibody A00547 for human tissue, and we are content with the WB results. The species of reactivity given in the datasheet says human, mouse, rat. Is it possible that the antibody can work on primate tissues as well?
Verified Customer
Verified customer
Asked: 2020-02-03
Answer
The anti-NEDD8 antibody (A00547) has not been validated for cross reactivity specifically with primate tissues, but there is a good chance of cross reactivity. We have an innovator award program that if you test this antibody and show it works in primate you can get your next antibody for free. Please contact me if I can help you with anything.
Boster Scientific Support
Answered: 2020-02-03
