NEDD8 Recombinant Rabbit Monoclonal Antibody [JF69-09]
Catalog# ET1702-84
NEDD8 Recombinant Rabbit Monoclonal Antibody [JF69-09]
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WB
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IF-Cell
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IF-Tissue
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IHC-P
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IP
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Human
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Mouse
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Rat
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Green monkey
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unconjugated
Overview
Product Name
NEDD8 Recombinant Rabbit Monoclonal Antibody [JF69-09]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Synthetic peptide within Human NEDD8 aa 12-57 / 81.
Species Reactivity
Human, Mouse, Rat, Green monkey
Validated Applications
WB, IF-Cell, IF-Tissue, IHC-P, IP
Molecular Weight
Predicted band size: 9 kDa
Positive Control
COS-1 cell lysate, HeLa cell lysate, Raji cell lysate, HepG2 cell lysate, K-562 cell lysate, RAW264.7 cell lysate, NIH/3T3 cell lysate, PC-12 cell lysate, C6 cell lysate, HepG2, RAW264.7, human lung carcinoma tissue, human breast carcinoma tissue, human testis tissue, rat bladder tissue, mouse colon tissue, mouse heart tissue, mouse lung tissue, mouse skeletal muscle tissue, mouse spleen tissue, rat brain tissue, rat colon tissue, rat liver tissue, rat lung tissue, rat skeletal muscle tissue, rat testis tissue.
Conjugation
unconjugated
Clone Number
JF69-09
RRID
Product Features
Form
Liquid
Concentration
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.
Storage Buffer
1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
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WB
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1:500-1:1,000
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IF-Cell
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1:50-1:200
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IF-Tissue
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1:50-1:200
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IHC-P
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1:50-1:1,000
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IP
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1:50
Target
Function
NEDD8 is a ubiquitin (Ub)-like molecule that shares 80% homology with ubiquitin, a protein known to modify and target various proteins for proteolytic degradation. NEDD8 and the corresponding yeast homolog Rub1 are activated by the E1 ubiquitin activating enzyme UBA2, which forms isopeptide linkages between thio esters. Similar to the Ub-mediated proteolytic pathway, NEDD8 is covalently coupled to an E3 Ub ligase by the E2 conjugating enzyme, Ubc12. NEDD8 preferentially associates with the CUL-2 protein in the E3 ligase complex CBCVHL, which consists of cullin proteins associating with elongin B/C and the tumor suppressor F-box protein, von Hippel Lindau (VHL). NEDD8 is predominantly localized to the nucleus and is highly expressed in adult heart and skeletal muscle. In vitro studies indicate that NEDD8 tetramers are also able to bind to the 26S Proteasome, and that they assemble into complexes with conserved Ub-like moieties, suggesting that NEDD8 may regulate proteolysis of intracellular proteins similar to other Ub-mediated pathways.
Background References
1. Lu R et al. COPS5 amplification and overexpression confers tamoxifen-resistance in ERa-positive breast cancer by degradation of NCoR. Nat Commun 7:12044 (2016).
2. Yu C et al. CRL4-DCAF1 ubiquitin E3 ligase directs protein phosphatase 2A degradation to control oocyte meiotic maturation. Nat Commun 6:8017 (2015).
Sequence Similarity
Belongs to the ubiquitin family.
Tissue Specificity
Highly expressed in heart, skeletal muscle, spleen, thymus, prostate, testis, ovary, colon and leukocytes.
Post-translational Modification
Cleavage of precursor form by UCHL3 or SENP8 is necessary for function.
Subcellular Location
Nucleus.
Synonyms
FLJ43224 antibody
MGC104393 antibody
MGC125896 antibody
MGC125897 antibody
NED8 antibody
NEDD 8 antibody
NEDD-8 antibody
Nedd8 antibody
NEDD8_HUMAN antibody
Neddylin antibody
ExpandFLJ43224 antibody
MGC104393 antibody
MGC125896 antibody
MGC125897 antibody
NED8 antibody
NEDD 8 antibody
NEDD-8 antibody
Nedd8 antibody
NEDD8_HUMAN antibody
Neddylin antibody
Neural precursor cell expressed developmentally down regulated 8 antibody
Neural precursor cell expressed developmentally down regulated gene 8 antibody
Neural precursor cell expressed developmentally down-regulated protein 8 antibody
Rub1 antibody
Ubiquitin like protein Nedd 8 antibody
Ubiquitin like protein Nedd8 antibody
Ubiquitin-like protein Nedd8 antibody
CollapseImages
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Western blot analysis of NEDD8 on different lysates with Rabbit anti-NEDD8 antibody (ET1702-84) at 1/1,000 dilution.
Lane 1: COS-1 cell lysate
Lane 2: HeLa cell lysate
Lane 3: Raji cell lysate
Lane 4: HepG2 cell lysate
Lane 5: K-562 cell lysate
Lane 6: RAW264.7 cell lysate
Lane 7: NIH/3T3 cell lysate
Lane 8: PC-12 cell lysate
Lane 9: C6 cell lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 9 kDa
Observed band size: 9-90 kDa
Exposure time: 3 minutes;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1702-84) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
ICC staining of NEDD8 in HepG2 cells (red). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1702-84, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®594 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
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Immunocytochemistry analysis of RAW264.7 cells labeling NEDD8 with Rabbit anti-NEDD8 antibody (ET1702-84) at 1/100 dilution.
Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-NEDD8 antibody (ET1702-84) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
Immunohistochemical analysis of paraffin-embedded human lung carcinoma tissue using anti-NEDD8 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1702-84, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using anti-NEDD8 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1702-84, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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Immunohistochemical analysis of paraffin-embedded human testis tissue with Rabbit anti-NEDD8 antibody (ET1702-84) at 1/400 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1702-84) at 1/400 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded rat bladder tissue with Rabbit anti-NEDD8 antibody (ET1702-84) at 1/400 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1702-84) at 1/400 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded mouse colon tissue with Rabbit anti-NEDD8 antibody (ET1702-84) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1702-84) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded mouse heart tissue with Rabbit anti-NEDD8 antibody (ET1702-84) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1702-84) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded mouse lung tissue with Rabbit anti-NEDD8 antibody (ET1702-84) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1702-84) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded mouse skeletal muscle tissue with Rabbit anti-NEDD8 antibody (ET1702-84) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1702-84) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded mouse spleen tissue with Rabbit anti-NEDD8 antibody (ET1702-84) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1702-84) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-NEDD8 antibody (ET1702-84) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1702-84) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded rat colon tissue with Rabbit anti-NEDD8 antibody (ET1702-84) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1702-84) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded rat liver tissue with Rabbit anti-NEDD8 antibody (ET1702-84) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1702-84) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded rat lung tissue with Rabbit anti-NEDD8 antibody (ET1702-84) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1702-84) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded rat skeletal muscle tissue with Rabbit anti-NEDD8 antibody (ET1702-84) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1702-84) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded rat testis tissue with Rabbit anti-NEDD8 antibody (ET1702-84) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1702-84) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
Citation
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Fibroblast-targeted MLN4924 suppresses autoimmune myocarditis by dually modulating IL-17A and TGF-β pathways
Journal: International Immunopharmacology
DOI: 10.1016/j.intimp.2026.116206
IF: 4.7
Application: IHC
Reactivity: Mouse
Publish date: 2026 Jan