重组Anti-NF-kB p65抗体[EP2161Y] (ab76311)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP2161Y] to NF-kB p65
- Suitable for: WB, ICC/IF, Flow Cyt (Intra)
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
概述
-
产品名称
Anti-NF-kB p65抗体[EP2161Y]
参阅全部 NF-kB p65 一抗 -
描述
兔单克隆抗体[EP2161Y] to NF-kB p65 -
宿主
Rabbit -
特异性
ab76311 detects both phosphorylated and non-phosphorylated versions of human NF-kB p65. -
经测试应用
适用于: WB, ICC/IF, Flow Cyt (Intra)more details
不适用于: IHC-P or IP -
种属反应性
与反应: Human -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
-
阳性对照
- WB: HeLa, Daudi and Jurkat cell lysates. Flow Cyt (intra): HeLa and HT-29 cells. ICC: HT-29 cells
-
常规说明
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.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
性能
-
形式
Liquid -
存放说明
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
存储溶液
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.5% BSA -
Concentration information loading...
-
纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EP2161Y -
同种型
IgG -
研究领域
相关产品
-
Alternative Versions
-
Assay kits
-
Isotype control
-
Positive Controls
-
Recombinant Protein
-
Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab76311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 |
---|---|---|
WB | (1) |
1/1000 - 1/5000. Detects a band of approximately 70 kDa (predicted molecular weight: 65 kDa).
|
ICC/IF |
1/100 - 1/500.
|
|
Flow Cyt (Intra) |
1/1000.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
|
说明 |
---|
WB
1/1000 - 1/5000. Detects a band of approximately 70 kDa (predicted molecular weight: 65 kDa). |
ICC/IF
1/100 - 1/500. |
Flow Cyt (Intra)
1/1000. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
|
靶标
-
功能
NF-kappa-B is a pleiotropic transcription factor which is present in almost all cell types and is involved in many biological processed such as inflammation, immunity, differentiation, cell growth, tumorigenesis and apoptosis. NF-kappa-B is a homo- or heterodimeric complex formed by the Rel-like domain-containing proteins RELA/p65, RELB, NFKB1/p105, NFKB1/p50, REL and NFKB2/p52 and the heterodimeric p65-p50 complex appears to be most abundant one. The dimers bind at kappa-B sites in the DNA of their target genes and the individual dimers have distinct preferences for different kappa-B sites that they can bind with distinguishable affinity and specificity. Different dimer combinations act as transcriptional activators or repressors, respectively. NF-kappa-B is controlled by various mechanisms of post-translational modification and subcellular compartmentalization as well as by interactions with other cofactors or corepressors. NF-kappa-B complexes are held in the cytoplasm in an inactive state complexed with members of the NF-kappa-B inhibitor (I-kappa-B) family. In a conventional activation pathway, I-kappa-B is phosphorylated by I-kappa-B kinases (IKKs) in response to different activators, subsequently degraded thus liberating the active NF-kappa-B complex which translocates to the nucleus. NF-kappa-B heterodimeric p65-p50 and p65-c-Rel complexes are transcriptional activators. The NF-kappa-B p65-p65 complex appears to be involved in invasin-mediated activation of IL-8 expression. The inhibitory effect of I-kappa-B upon NF-kappa-B the cytoplasm is exerted primarily through the interaction with p65. p65 shows a weak DNA-binding site which could contribute directly to DNA binding in the NF-kappa-B complex. Associates with chromatin at the NF-kappa-B promoter region via association with DDX1. -
序列相似性
Contains 1 RHD (Rel-like) domain. -
结构域
the 9aaTAD motif is a transactivation domain present in a large number of yeast and animal transcription factors. -
翻译后修饰
Ubiquitinated, leading to its proteasomal degradation. Degradation is required for termination of NF-kappa-B response.
Monomethylated at Lys-310 by SETD6. Monomethylation at Lys-310 is recognized by the ANK repeats of EHMT1 and promotes the formation of repressed chromatin at target genes, leading to down-regulation of NF-kappa-B transcription factor activity. Phosphorylation at Ser-311 disrupts the interaction with EHMT1 without preventing monomethylation at Lys-310 and relieves the repression of target genes.
Phosphorylation at Ser-311 disrupts the interaction with EHMT1 and promotes transcription factor activity (By similarity). Phosphorylation on Ser-536 stimulates acetylation on Lys-310 and interaction with CBP; the phosphorylated and acetylated forms show enhanced transcriptional activity.
Reversibly acetylated; the acetylation seems to be mediated by CBP, the deacetylation by HDAC3. Acetylation at Lys-122 enhances DNA binding and impairs association with NFKBIA. Acetylation at Lys-310 is required for full transcriptional activity in the absence of effects on DNA binding and NFKBIA association. Acetylation can also lower DNA-binding and results in nuclear export. Interaction with BRMS1 promotes deacetylation of 'Lys-310'. -
细胞定位
Nucleus. Cytoplasm. Nuclear, but also found in the cytoplasm in an inactive form complexed to an inhibitor (I-kappa-B). Colocalized with RELA in the nucleus upon TNF-alpha induction. - Information by UniProt
-
数据库链接
- Entrez Gene: 5970 Human
- Omim: 164014 Human
- SwissProt: Q04206 Human
- Unigene: 502875 Human
-
别名
- Avian reticuloendotheliosis viral (v rel) oncogene homolog A antibody
- MGC131774 antibody
- NF kappa B p65delta3 antibody
see all
图片
-
Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: NF-κB p65 knockout HAP1 cell lysate (20 µg)
Lane 3: HeLa cell lysate (20 µg)
Lane 4: A431 cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab76311 observed at 70 kDa. Red - ab8245 loading control, observed at 37 kDa.
ab76311 was shown to specifically react with NF-κB p65 in wild-type HAP1 cells along with additional cross-reactive bands. No band was observed when NF-κB p65 knockout samples were used. Wild-type and NF-κB p65 knockout samples were subjected to SDS-PAGE. ab76311 (NF-κB p65) and ab8245 (loading control to GAPDH) were both diluted 1/1000 and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 h at room temperature before imaging. -
Immunocytochemistry/Immunofluorescence analysis HT-29 (human colorectal adenocarcinoma) labelling NF-kB p65 with purified ab76311 at 1/500. Cells were fixed with 4% PFA and permeabilized with 0.1% Triton X-100. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody (Ab150077). Nuclei counterstained with DAPI (blue).
Control: PBS only
-
Flow Cytometry analysis of HT-29 (Human colorectal adenocarcinoma epithelial cell) cells labeling NF-kB p65 with purified ab76311 at 1/1000 dilution (1 µg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
-
All lanes : Anti-NF-kB p65 antibody [EP2161Y] (ab76311) at 1/20000 dilution
Lane 1 : HeLa cell lysate
Lane 2 : Daudi cell lysate
Lane 3 : Jurkat cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution
Predicted band size: 65 kDa
Observed band size: 70 kDa why is the actual band size different from the predicted? -
Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling NF-kB p65 with purified ab76311 at 1/1000 dilution (1 µg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
实验方案
数据表及文件
-
SDS download
-
Datasheet download
文献 (19)
ab76311 被引用在 19 文献中.
- Liu H et al. MicroRNA-421 attenuates macrophage-mediated inflammation by inhibiting PDCD4 in vitro. Mol Med Rep 24:N/A (2021). PubMed: 34036390
- Hu M et al. Smad7 attenuates TGF-β-mediated aging-related hypofunction of submandibular glands. Exp Biol Med (Maywood) 246:1269-1273 (2021). PubMed: 33641444
- Liu Y et al. The TGF-β/miR-31/CEACAM1-S axis inhibits CD4+ CD25+ Treg differentiation in systemic lupus erythematosus. Immunol Cell Biol 99:697-710 (2021). PubMed: 33655578
- Sotillo WS et al. Breast cancer cell line toxicity of a flavonoid isolated from Baccharis densiflora. BMC Complement Med Ther 21:188 (2021). PubMed: 34215226
- Li Q et al. Carminic acid supplementation protects against fructose-induced kidney injury mainly through suppressing inflammation and oxidative stress via improving Nrf-2 signaling. Aging (Albany NY) 13:10326-10353 (2021). PubMed: 33819919