Anti-NEDD8 Antibody Picoband™

Product Name

Anti-NEDD8 Antibody Picoband™

View all NEDD8 Antibodies

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.

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

Applications

A00547 is guaranteed for Flow Cytometry, IF, IHC, ICC, WB Boster Guarantee

Observed Molecular Weight

12 kDa

Calculated molecular weight

9.072kDa

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.

View more details

Innovating Scientists Reward

If you are the first to review this product, or if you have results for a special sample, species or application this product is not validated in, share your results with us and receive product credits you can use towards any Boster products! Applicable to all scientists worldwide.

Submit A Review

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

Validation Images & Assay Conditions

Click image to see more details

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 50ug of sample under reducing conditions.
Lane 1: rat testis cell lysates,
Lane 2: mouse thymus tissue lysates,
Lane 3: mouse brain tissue lysates,
Lane 4: HELA 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-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 1:10000 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 12KD. The expected band size for NEDD8 is at 9KD.

Click image to see more details

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.

Click image to see more details

Figure 3. Flow Cytometry analysis of A431 cells using anti-NEDD8 antibody (A00547).
Overlay histogram showing A431 cells stained with A00547 (Blue line).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 (Red line) was also used as a control.

Click image to see more details

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.

Gene/Protein Information For NEDD8 (Source: Uniprot.org, NCBI)

Alternative Names

FLJ43224; MGC104393; MGC125896; MGC125897; NEDD8; NEDD-8; Neddylin; Neural precursor cell expressed developmentally down-regulated protein 8; neural precursor cell expressed, developmentally down-regulated 8; Rub1; Ubiquitin-like protein Nedd8 NEDD8 NEDD-8 NEDD8 ubiquitin like modifier NEDD8|neddylin|neural precursor cell expressed, developmentally down-regulated 8|ubiquitin-like protein Nedd8

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].

Take me to the NEDD8 Infographic

Here are featured tools and databases that you might find useful.

• Boster's Pathways Library
• Protein Databases
• Bioscience Research Protocol Resources
• Data Processing & Analysis Software
• Photo Editing Software
• Scientific Literature Resources
• Research Paper Management Tools
• Molecular Biology Software
• Primer Design Tools
• Bioinformatics Tools
• Phylogenetic Tree Analysis