Rabbit Recombinant Monoclonal NEDD8 antibody. Suitable for IP, WB, ICC/IF, Flow Cyt (Intra), IHC-P and reacts with Human, Rat, Mouse samples. Cited in 42 publications.
IgG
Rabbit
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 50% Glycerol (glycerin, glycerine), 49% PBS, 0.05% BSA
Liquid
Monoclonal
IP | WB | ICC/IF | Flow Cyt (Intra) | IHC-P | |
---|---|---|---|---|---|
Human | Tested | Tested | Tested | Tested | Tested |
Mouse | Expected | Tested | Expected | Expected | Expected |
Rat | Expected | Tested | Expected | Expected | Expected |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/20 - 1/50 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Rat, Mouse | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info 1/1000 - 1/10000 | Notes - |
Species Mouse | Dilution info 1/1000 - 1/10000 | Notes - |
Species Human | Dilution info 1/1000 - 1/10000 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/100 - 1/250 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Rat, Mouse | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/300 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Rat, Mouse | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/100 - 1/500 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Rat, Mouse | Dilution info Use at an assay dependent concentration. | Notes - |
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Ubiquitin-like protein which plays an important role in cell cycle control and embryogenesis via its conjugation to a limited number of cellular proteins, such as cullins or p53/TP53 (PubMed:10318914, PubMed:10597293, PubMed:11953428, PubMed:14690597, PubMed:15242646, PubMed:9694792, PubMed:38605244). Attachment of NEDD8 to cullins is critical for the recruitment of E2 to the cullin-RING-based E3 ubiquitin-protein ligase complex, thus facilitating polyubiquitination and proteasomal degradation of cyclins and other regulatory proteins (PubMed:10318914, PubMed:10597293, PubMed:11953428, PubMed:20688984, PubMed:9694792, PubMed:38605244). Attachment of NEDD8 to p53/TP53 inhibits p53/TP53 transcriptional activity (PubMed:15242646). Covalent attachment to its substrates requires prior activation by the E1 complex UBE1C-APPBP1 and linkage to the E2 enzyme UBE2M (PubMed:14690597).
Ubiquitin-like protein NEDD8, Neddylin, Neural precursor cell expressed developmentally down-regulated protein 8, NEDD-8, NEDD8
Rabbit Recombinant Monoclonal NEDD8 antibody. Suitable for IP, WB, ICC/IF, Flow Cyt (Intra), IHC-P and reacts with Human, Rat, Mouse samples. Cited in 42 publications.
IgG
Rabbit
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 50% Glycerol (glycerin, glycerine), 49% PBS, 0.05% BSA
Liquid
Monoclonal
Y297
Affinity purification Protein A
This antibody detects NEDD8 bound to cullins proteins (neddylated cullins) as well as free NEDD8.
Blue Ice
1-2 weeks
+4°C
-20°C
Upon delivery aliquot
Stable for 12 months at -20°C
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
This product is a recombinant monoclonal antibody, which offers several advantages including:
For more information, read more on recombinant antibodies.
This supplementary information is collated from multiple sources and compiled automatically.
NEDD8 also known as neural precursor cell expressed developmentally downregulated protein 8 is a small ubiquitin-like protein with a molecular weight of approximately 9 kDa. It is expressed in various tissues prominently in the brain and muscles. NEDD8 conjugates to target proteins modifying their functions by altering their stability activity or location. It plays a role in the regulation of protein degradation.
The NEDD8 protein is part of a modification system that activates cullin-RING ligase complexes. This activation promotes the ubiquitination and subsequent proteasomal degradation of targeted proteins. As a post-translational modification system similar to ubiquitin NEDD8 is important for maintaining protein homeostasis by ensuring timely degradation and turnover of cellular proteins. These processes impact cell cycle progression signal transduction and stress responses.
NEDD8 is a central player in the ubiquitin-proteasome system an essential pathway for protein turnover. It is particularly important in the NEDDylation pathway where it regulates protein functions by transiently attaching to substrate proteins. Through these processes NEDD8 interacts closely with proteins like cullins and ubiquitin affecting cellular responses and maintaining cellular integrity under different conditions.
NEDD8 influences several pathological states. Aberrant NEDD8 conjugation can lead to cancer by disrupting the controlled degradation of proteins involved in cell cycle regulation such as p27 and p21. Additionally improper NEDDylation has links to neurodegenerative diseases like Alzheimer's as it affects proteins related to synaptic function and neuronal survival. Understanding the role of NEDD8 in these diseases aids in the development of potential therapeutic strategies.
We have tested this species and application combination and it works. It is covered by our product promise.
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our product promise.
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All lanes: Western blot - Anti-NEDD8 antibody [Y297] (ab81264) at 1/1000 dilution
Lane 1: C6 Cell Lysate at 20 µg
Lane 2: Raw264.7 Cell Lysate at 20 µg
Lane 3: PC-12 Cell Lysate at 20 µg
Lane 4: 3T3 Cell Lysate at 20 µg
All lanes: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution
Predicted band size: 9 kDa
Immunohistochemical staining of paraffin embedded human cervical carcinoma with purified ab81264 at a working dilution of 1/400. The secondary antibody used is Goat Anti-Rabbit IgG H&L (HRP) ab97051, a goat anti-rabbit IgG (H&L) at a dilution of 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
Immunofluorescence staining of HeLa cells with purified ab81264 at a working dilution of 1/100, counter-stained with DAPI. The secondary antibody was Alexa Fluor® 488 goat anti-rabbit (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077), used at a dilution of 1/1000. Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 (Alexa Fluor® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified ab81264 was used at a dilution of 1/500 followed by an Alexa Fluor® 594 goat anti-mouse antibody (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) at a dilution of 1/500. For negative control 2, Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor® 488 goat anti-rabbit antibody (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at a dilution of 1/400.
ab81264 (purified) at 1/20 immunoprecipitating NEDD8 in 10 μg K562 cell lysate (Lanes 1 and 2, observed at 10 and 90 kDa). Lane 3 - Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730). For western blotting, VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366), was used for detection at 1/10,000 dilution. Blocking buffer and concentration: 5% NFDM/TBST Dilution buffer and concentration: 5% NFDM/TBST
All lanes: Immunoprecipitation - Anti-NEDD8 antibody [Y297] (ab81264)
Predicted band size: 9 kDa
Intracellular Flow Cytometry analysis of A-673 (Human muscle Ewing's Sarcoma) cells labeling NEDD8 (red) with purified ab81264 at a 1/300 dilution (1ug/mL). Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A goat anti rabbit IgG (Alexa Fluorr®488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) was used as the secondary antibody at a 1/2000 dilution. Black - Rabbit monoclonal IgG (Black) (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730). Blue (unlabeled control) - Cell without incubation with primary antibody and secondary antibody (Blue).
Western blot analysis of immunoprecipitation pellet from HeLa cell lysate using unpurified ab81264 at 1/500 dilution. Band at 90 kDa shows Neddylated cullins, 50 kDa the IgG fraaction and 9 kDa Free NEDD8.
All lanes: Immunoprecipitation - Anti-NEDD8 antibody [Y297] (ab81264)
Predicted band size: 9 kDa
Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
The band at 12kDa is free NEDD8, and at 90kDa is Neddylated cullins.
All lanes: Western blot - Anti-NEDD8 antibody [Y297] (ab81264) at 1/1000 dilution
All lanes: PC-12 whole cell lysate at 10 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 9 kDa
Observed band size: 12 kDa, 90 kDa
Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
The band at 12kDa is free NEDD8, and at 90kDa is Neddylated cullins.
All lanes: Western blot - Anti-NEDD8 antibody [Y297] (ab81264) at 1/1000 dilution
All lanes: K562 whole cell lysate at 10 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 9 kDa
Observed band size: 12 kDa, 90 kDa
Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
The band at 12kDa is free NEDD8, and at 90kDa is Neddylated cullins.
All lanes: Western blot - Anti-NEDD8 antibody [Y297] (ab81264) at 1/1000 dilution
All lanes: HEK293 cell lysate at 10 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 9 kDa
Observed band size: 12 kDa, 90 kDa
Other Bands: 90kDa Neddylated cullins
All lanes: Western blot - Anti-NEDD8 antibody [Y297] (ab81264) at 1/500 dilution
Lane 1: HeLa cell lysate at 10 µg
Lane 2: K562 cell lysate at 10 µg
All lanes: HRP labelled goat anti-rabbit at 1/2000 dilution
Predicted band size: 9 kDa
Observed band size: 9 kDa
Unpurified ab81264, at 1/100 dilution, staining NEDD8 in human lung carcinoma tissue by Immunohistochemistry using formalin-fixed, paraffin-embedded section.
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