WB
Western blot analysis of NEDD8 on different lysates with Rabbit anti-NEDD8 antibody 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. 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 at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.ICC/IF
Immunocytochemistry analysis of RAW264.7 cells labeling NEDD8 with Rabbit anti-NEDD8 antibody 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 at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.IHC
Immunohistochemical analysis of paraffin-embedded rat bladder tissue with Rabbit anti-NEDD8 antibody at 1/400 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) 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 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.IHC
Immunohistochemical analysis of paraffin-embedded rat colon tissue with Rabbit anti-NEDD8 antibody at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) 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 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.| Product Name | NEDD8 Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Product description | 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. |
| Immunogen | Synthetic peptide within Human NEDD8 aa 12-57 / 81. |
| Clonality | monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Recombinant rabbit |
| Tested Applications | ICC/IFIHCWB |
| WB:1:500-1:1000 ICC:1:50-1:200 IHC:1:50-1:1000 |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | NEDD8 |
|---|---|
| Gene Synonyms | NEDD-8 |
| Gene Full Name | NEDD8 ubiquitin like modifier |
| Alternative Names | FLJ43224 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 |
| Molecular Weight(MW) | 9kDa(Observed band size: 9-90kDa) |
| Cellular Localization | Nucleus. |
WB
Western blot analysis of NEDD8 on different lysates with Rabbit anti-NEDD8 antibody 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. 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 at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
ICC/IF
Immunocytochemistry analysis of RAW264.7 cells labeling NEDD8 with Rabbit anti-NEDD8 antibody 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 at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.
IHC
Immunohistochemical analysis of paraffin-embedded rat bladder tissue with Rabbit anti-NEDD8 antibody at 1/400 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) 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 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.
IHC
Immunohistochemical analysis of paraffin-embedded rat colon tissue with Rabbit anti-NEDD8 antibody at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) 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 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.| Positive Control | HUVEC, MCF-7, Hela, HepG2, human liver tissue, mouse liver tissue, mouse brain tissue, human lung tissue. |
|---|---|
| Application Notes | WB:1:500-1:1000 ICC:1:50-1:200 IHC:1:50-1:1000 |
| Form | Liquid |
|---|---|
| Storage Instructions | Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | 1*TBS (pH7.4), 1%BSA, 40%Glycerol. Preservative: 0.05% Sodium Azide. |
