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Anti-NEDD8 Antibody

Catalog Number: orb389427

DispatchUsually dispatched within 3-4 weeks
$ 210.00
Catalog Numberorb389427
CategoryAntibodies
DescriptionAnti-NEDD8 Antibody
Species/HostRabbit
ClonalityPolyclonal
Tested applicationsFC, ICC, IF, IHC, WB
ReactivityHuman, Mouse, Rat
IsotypeRabbit IgG
ImmunogenA synthetic peptide corresponding to a sequence in the middle region of human NEDD8, identical to the related mouse and rat sequences.
Antibody TypePrimary Antibody
ConcentrationAdding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
Form/AppearanceLyophilized
ConjugationUnconjugated
MW12 kDa
UniProt IDQ15843
StorageMaintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
Alternative namesNEDD8; Neddylin; Neural precursor cell expressed d
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NoteFor research use only
Application notesWestern blot, 0.1-0.5μg/ml, Human, Mouse, Rat Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Human Immunocytochemistry/Immunofluorescence, 2μg/ml, Human Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human. Add 0.2ml of distilled water will yield a concentration of 500ug/ml
Expiration Date12 months from date of receipt.
Anti-NEDD8 Antibody

Flow Cytometry analysis of A431 cells using anti-NEDD8 antibody. Overlay histogram showing A431 cells (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 (1 µg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (5-10 µg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 µg/1x10^6) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

Anti-NEDD8 Antibody

IF analysis of NEDD8 using anti-NEDD8 antibody. NEDD8 was detected in immunocytochemical section of A431 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2 µg/mL rabbit anti-NEDD8 Antibody overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG 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.

Anti-NEDD8 Antibody

IHC analysis of NEDD8 using anti-NEDD8 antibody. 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 1 µg/ml rabbit anti-NEDD8 Antibody 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) with DAB as the chromogen.

Anti-NEDD8 Antibody

Western blot analysis of NEDD8 using anti-NEDD8 antibody. 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 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 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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