重组Anti-Rel B抗体[EP614Y] (ab33907)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP614Y] to Rel B
- Suitable for: Flow Cyt (Intra), ICC/IF, WB, IP
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
概述
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产品名称
Anti-Rel B抗体[EP614Y]
参阅全部 Rel B 一抗 -
描述
兔单克隆抗体[EP614Y] to Rel B -
宿主
Rabbit -
经测试应用
适用于: Flow Cyt (Intra), ICC/IF, WB, IPmore details
不适用于: ChIP or IHC-P -
种属反应性
与反应: Human -
免疫原
Synthetic peptide within Human Rel B aa 550-650 (C terminal). The exact sequence is proprietary.
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阳性对照
- WB: HeLa, Raji and Daudi cell lysate. IP: Daudi cell lysate, Raji whole cell lysate ICC/IF: Raji cells. Flow Cyt (intra): Raji cells.
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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
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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), 0.05% BSA -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EP614Y -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
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Recombinant Protein
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab33907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) |
1/70.
For unpurified, use 1/1000. ab172730 - Rabbit monoclonal IgG is suitable for use as an isotype control with this antibody. |
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ICC/IF |
1/100.
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WB |
1/2000 - 1/20000. Predicted molecular weight: 62 kDa.
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IP |
1/40 - 1/50.
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说明 |
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Flow Cyt (Intra)
1/70. For unpurified, use 1/1000. ab172730 - Rabbit monoclonal IgG is suitable for use as an isotype control with this antibody. |
ICC/IF
1/100. |
WB
1/2000 - 1/20000. Predicted molecular weight: 62 kDa. |
IP
1/40 - 1/50. |
靶标
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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. 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 RelB-p50 and RelB-p52 complexes are transcriptional activators. RELB neither associates with DNA nor with RELA/p65 or REL. Stimulates promoter activity in the presence of NFKB2/p49. -
序列相似性
Contains 1 RHD (Rel-like) domain. -
结构域
Both N- and C-terminal domains are required for transcriptional activation. -
翻译后修饰
Phosphorylation at 'Thr-103' and 'Ser-573' is followed by proteasomal degradation. -
细胞定位
Nucleus. Cytoplasm > cytoskeleton > centrosome. Co-localizes with NEK6 in the centrosome. - Information by UniProt
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数据库链接
- Entrez Gene: 5971 Human
- Omim: 604758 Human
- SwissProt: Q01201 Human
- Unigene: 654402 Human
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别名
- I REL antibody
- I-Rel antibody
- IREL antibody
see all
图片
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All lanes : Anti-Rel B antibody [EP614Y] (ab33907) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : RELB knockout HeLa cell lysate
Lane 3 : Raji cell lysate
Lane 4 : LnCap cell lysate
Lysates/proteins at 40 µg per lane.
Performed under reducing conditions.
Predicted band size: 62 kDa
Observed band size: 70 kDa why is the actual band size different from the predicted?Lanes 1- 4: Merged signal (red and green). Green - ab33907 observed at 70 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab33907 was shown to react with Rel B in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265948 (knockout cell lysate ab257635) was used. Wild-type HeLa and RELB knockout HeLa cell lysates were subjected to SDS-PAGE. ab33907 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at a 1 in 1000 Dilution and a 1 in 20000 dilution respectively. 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 in 20000 dilution for 1 hour at room temperature before imaging.
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All lanes : Anti-Rel B antibody [EP614Y] (ab33907) at 1/2000 dilution (purified)
Lane 1 : Raji cell lysate
Lane 2 : Daudi cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 62 kDa
Observed band size: 70 kDa why is the actual band size different from the predicted?Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST -
Immunofluorescence staining of Raji cells with purified ab33907 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 ab33907 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.
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ab33907 (purified) at 1/20 immunoprecipitating Rel B in 10 μg Daudi cell lysate (Lanes 1 and 2, observed at 70 kDa). Lane 3 - Rabbit monoclonal IgG (ab172730). For western blotting, HRP Veriblot for IP (ab131366) was used for detection (1/1000). Blocking buffer and concentration: 5% NFDM/TBST Dilution buffer and concentration: 5% NFDM/TBST
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Rel B was immunoprecipitated from 0.35 mg of Raji (human Burkitt's lymphoma B lymphocyte), whole cell lysate with ab33907 at 1/30 dilution. Western blot was performed from the immunoprecipitate using 33907 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.
Lane 1: Raji whole cell lysate 10 μg (Input).
Lane 2: ab33907 IP in Raji whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab33907 in Raji whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 180 seconds.
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Overlay histogram showing Raji cells fixed in 80% methanol and stained with purified ab33907 at a dilution of 1/70 (red line). The secondary antibody used was Alexa Fluorr® 488 goat anti-rabbit at a dilution of 1/500. Rabbit monoclonal IgG was used as an isotype control (black line) and cells incubated in the absence of both primary and secondary antibody were used as a negative control (blue line).
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Anti-Rel B antibody [EP614Y] (ab33907) at 1/20000 dilution (unpurified) + Raji Cell Lysate
Predicted band size: 62 kDa
Observed band size: 62 kDa -
Overlay histogram showing Raji cells stained with unpurified ab33907 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab33907, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluorr® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1µg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in Raji cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
实验方案
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
数据表及文件
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SDS download
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Datasheet download
文献 (3)
ab33907 被引用在 3 文献中.
- Yuan J et al. MiRNA-223-3p Affects Mantle Cell Lymphoma Development by Regulating the CHUK/NF-?B2 Signaling Pathway. Onco Targets Ther 14:1553-1564 (2021). PubMed: 33688203
- Wang RW et al. Aneuploid senescent cells activate NF-κB to promote their immune clearance by NK cells. EMBO Rep 22:e52032 (2021). PubMed: 34105235
- Chen X et al. The NF-kappaB factor RelB and histone H3 lysine methyltransferase G9a directly interact to generate epigenetic silencing in endotoxin tolerance. J Biol Chem 284:27857-65 (2009). IP ; Human . PubMed: 19690169