重组Anti-NEDD8抗体[Y297] - BSA and Azide free (ab220816)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [Y297] to NEDD8 - BSA and Azide free
- Suitable for: Flow Cyt (Intra), WB, IP, IHC-P, ICC/IF
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
概述
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产品名称
Anti-NEDD8抗体[Y297] - BSA and Azide free
参阅全部 NEDD8 一抗 -
描述
兔单克隆抗体[Y297] to NEDD8 - BSA and Azide free -
宿主
Rabbit -
特异性
This antibody detects NEDD8 bound to cullins proteins (neddylated cullins) as well as free NEDD8.
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经测试应用
适用于: Flow Cyt (Intra), WB, IP, IHC-P, ICC/IFmore details -
种属反应性
与反应: Mouse, Rat, Human -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- Hela and K562 cell lysates and human lung carcinoma.
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常规说明
ab220816 is the carrier-free version of ab81264.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
性能
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形式
Liquid -
存放说明
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
存储溶液
pH: 7.20
Constituent: PBS -
无载体
是 -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
Y297 -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
- Alexa Fluor® 647 Anti-NEDD8 antibody [Y297] (ab200849)
- Alexa Fluor® 488 Anti-NEDD8 antibody [Y297] (ab201249)
- Alexa Fluor® 594 Anti-NEDD8 antibody [Y297] (ab207247)
- Alexa Fluor® 405 Anti-NEDD8 antibody [Y297] (ab207248)
- Alexa Fluor® 555 Anti-NEDD8 antibody [Y297] (ab207249)
- PE Anti-NEDD8 antibody [Y297] (ab305725)
- APC Anti-NEDD8 antibody [Y297] (ab305726)
- HRP Anti-NEDD8 antibody [Y297] (ab305727)
- Alexa Fluor® 568 Anti-NEDD8 antibody [Y297] (ab312568)
- Anti-NEDD8 antibody [Y297] (ab81264)
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Compatible Secondaries
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Conjugation kits
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Isotype control
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Recombinant Protein
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab220816于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。
应用 | Ab评论 | 说明 |
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Flow Cyt (Intra) |
Use at an assay dependent concentration.
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WB |
Use at an assay dependent concentration. Detects a band of approximately 9 kDa (predicted molecular weight: 9 kDa).
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IP |
Use at an assay dependent concentration.
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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ICC/IF |
Use at an assay dependent concentration.
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说明 |
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Flow Cyt (Intra)
Use at an assay dependent concentration. |
WB
Use at an assay dependent concentration. Detects a band of approximately 9 kDa (predicted molecular weight: 9 kDa). |
IP
Use at an assay dependent concentration. |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
ICC/IF
Use at an assay dependent concentration. |
靶标
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功能
Ubiquitin-like protein which plays an important role in cell cycle control and embryogenesis. Covalent attachment to its substrates requires prior activation by the E1 complex UBE1C-APPBP1 and linkage to the E2 enzyme UBE2M. Attachment of NEDD8 to cullins activates their associated E3 ubiquitin ligase activity, and thus promotes polyubiquitination and proteasomal degradation of cyclins and other regulatory proteins. -
组织特异性
Highly expressed in heart, skeletal muscle, spleen, thymus, prostate, testis, ovary, colon and leukocytes. -
序列相似性
Belongs to the ubiquitin family. -
翻译后修饰
Cleavage of precursor form by UCHL3 or SENP8 is necessary for function. -
细胞定位
Nucleus. Mainly nuclear. - Information by UniProt
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数据库链接
- Entrez Gene: 4738 Human
- Entrez Gene: 18002 Mouse
- Entrez Gene: 25490 Rat
- Omim: 603171 Human
- SwissProt: Q15843 Human
- SwissProt: P29595 Mouse
- SwissProt: Q71UE8 Rat
- Unigene: 531064 Human
see all -
别名
- FLJ43224 antibody
- MGC104393 antibody
- MGC125896 antibody
see all
图片
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Immunohistochemical staining of paraffin embedded human cervical carcinoma with purified ab81264 at a working dilution of 1/400. The secondary antibody used is 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab81264).
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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 (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with 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 (ab150120) at a dilution of 1/500. For negative control 2, ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor® 488 goat anti-rabbit antibody (ab150077) at a dilution of 1/400.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab81264).
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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 Fluor®488) (ab150077) was used as the secondary antibody at a 1/2000 dilution. Black - Rabbit monoclonal IgG (Black) (ab172730). Blue (unlabeled control) - Cell without incubation with primary antibody and secondary antibody (Blue).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab81264).
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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 (ab172730). For western blotting, HRP Veriblot for IP (ab131366) was used for detection at 1/1000 dilution. Blocking buffer and concentration: 5% NFDM/TBST Dilution buffer and concentration: 5% NFDM/TBST
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab81264).
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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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab81264).
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Unpurified ab81264, at 1/100 dilution, staining NEDD8 in human lung carcinoma tissue by Immunohistochemistry using formalin-fixed, paraffin-embedded section.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab81264).
实验方案
数据表及文件
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Datasheet download
Certificate of Compliance
文献 (4)
ab220816 被引用在 4 文献中.
- 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). Human . PubMed: 27375289
- Yu C et al. CRL4-DCAF1 ubiquitin E3 ligase directs protein phosphatase 2A degradation to control oocyte meiotic maturation. Nat Commun 6:8017 (2015). ICC/IF ; Mouse . PubMed: 26281983
- Godbersen JC et al. The Nedd8-activating enzyme inhibitor MLN4924 thwarts microenvironment-driven NF-?B activation and induces apoptosis in chronic lymphocytic leukemia B cells. Clin Cancer Res 20:1576-89 (2014). PubMed: 24634471
- van Knippenberg I et al. The transient nature of Bunyamwera orthobunyavirus NSs protein expression: effects of increased stability of NSs protein on virus replication. PLoS One 8:e64137 (2013). WB ; Human . PubMed: 23667701