重组Anti-NFkB p105 / p50抗体[E381] - BSA and Azide free (ab220803)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [E381] to NFkB p105 / p50 - BSA and Azide free
- Suitable for: WB, IHC-P
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
概述
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产品名称
Anti-NFkB p105 / p50抗体[E381] - BSA and Azide free
参阅全部 NFkB p105 / p50 一抗 -
描述
兔单克隆抗体[E381] to NFkB p105 / p50 - BSA and Azide free -
宿主
Rabbit -
特异性
This antibody will detect both forms: p50 and p105.
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经测试应用
适用于: WB, IHC-Pmore details
不适用于: Flow Cyt -
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
- WB: HeLa, HAP1, Jurkat, MCF-7 and PC-12 cell lysates. IHC-P: Human prostate carcinoma and bladder carcinoma tissues.
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常规说明
ab220803 is the carrier-free version of ab32360.
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
E381 -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
- HRP Anti-NFkB p105 / p50 antibody [E381] (ab195854)
- Alexa Fluor® 488 Anti-NFkB p105 / p50 antibody [E381] (ab311049)
- Alexa Fluor® 647 Anti-NFkB p105 / p50 antibody [E381] (ab311174)
- Alexa Fluor® 594 Anti-NFkB p105 / p50 antibody [E381] (ab311829)
- Alexa Fluor® 568 Anti-NFkB p105 / p50 antibody [E381] (ab313112)
- Alexa Fluor® 555 Anti-NFkB p105 / p50 antibody [E381] (ab313305)
- Anti-NFkB p105 / p50 antibody [E381] (ab32360)
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Compatible Secondaries
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Conjugation kits
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Isotype control
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Positive Controls
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab220803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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WB |
Use at an assay dependent concentration. Detects a band of approximately 50, 105 kDa (predicted molecular weight: 50 kDa).
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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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说明 |
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WB
Use at an assay dependent concentration. Detects a band of approximately 50, 105 kDa (predicted molecular weight: 50 kDa). |
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. |
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 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 RelB-p50 complexes are transcriptional activators. The NF-kappa-B p50-p50 homodimer is a transcriptional repressor, but can act as a transcriptional activator when associated with BCL3. NFKB1 appears to have dual functions such as cytoplasmic retention of attached NF-kappa-B proteins by p105 and generation of p50 by a cotranslational processing. The proteasome-mediated process ensures the production of both p50 and p105 and preserves their independent function, although processing of NFKB1/p105 also appears to occur post-translationally. p50 binds to the kappa-B consensus sequence 5'-GGRNNYYCC-3', located in the enhancer region of genes involved in immune response and acute phase reactions. In a complex with MAP3K8, NFKB1/p105 represses MAP3K8-induced MAPK signaling; active MAP3K8 is released by proteasome-dependent degradation of NFKB1/p105. -
序列相似性
Contains 7 ANK repeats.
Contains 1 death domain.
Contains 1 RHD (Rel-like) domain. -
结构域
The C-terminus of p105 might be involved in cytoplasmic retention, inhibition of DNA-binding, and transcription activation.
Glycine-rich region (GRR) appears to be a critical element in the generation of p50. -
翻译后修饰
While translation occurs, the particular unfolded structure after the GRR repeat promotes the generation of p50 making it an acceptable substrate for the proteasome. This process is known as cotranslational processing. The processed form is active and the unprocessed form acts as an inhibitor (I kappa B-like), being able to form cytosolic complexes with NF-kappa B, trapping it in the cytoplasm. Complete folding of the region downstream of the GRR repeat precludes processing.
Phosphorylation at 'Ser-903' and 'Ser-907' primes p105 for proteolytic processing in response to TNF-alpha stimulation. Phosphorylation at 'Ser-927' and 'Ser-932' are required for BTRC/BTRCP-mediated proteolysis.
Polyubiquitination seems to allow p105 processing.
S-nitrosylation of Cys-61 affects DNA binding. -
细胞定位
Nucleus. Cytoplasm. Nuclear, but also found in the cytoplasm in an inactive form complexed to an inhibitor. - Information by UniProt
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数据库链接
- Entrez Gene: 4790 Human
- Entrez Gene: 18033 Mouse
- Entrez Gene: 81736 Rat
- Omim: 164011 Human
- SwissProt: P19838 Human
- SwissProt: P25799 Mouse
- SwissProt: Q63369 Rat
- Unigene: 618430 Human
see all -
别名
- DKFZp686C01211 antibody
- DNA binding factor KBF1 antibody
- DNA binding factor KBF1 EBP1 antibody
see all
图片
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All lanes : Anti-NFkB p105 / p50 antibody [E381] (ab32360)
Lane 1 : Wild-type HAP1 cell lysate
Lane 2 : NFkB p105 / p50 knockout HAP1 cell lysate
Lane 3 : HeLa cell lysate
Lane 4 : Jurkat cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 50 kDaLanes 1 - 4: Merged signal (red and green). Green - ab32360 observed at 120, 50 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab32360 was shown to specifically react with NFkB p105 / p50 when NFkB p105 / p50 knockout samples were used. Wild-type and NFkB p105 / p50 knockout samples were subjected to SDS-PAGE. ab32360 and ab8245 (loading control to GAPDH) were diluted at 1/1000 and 1/10 000 respectively and 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 hour at room temperature before imaging.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32360).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human bladder carcinoma tissue labelling NFkB p105 / p50 with purified ab32360 at 1/250. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32360).
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All lanes : Anti-NFkB p105 / p50 antibody [E381] (ab32360) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : NFKB1 CRISPR/Cas9 edited HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 50 kDa
Observed band size: 105 kDa why is the actual band size different from the predicted?This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32360).
Lanes 1- 2: Merged signal (red and green). Green - ab32360 observed at 105 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab32360 was shown to react with NFkB p105 / p50 in wild-type HeLa cells in western blot. The band observed in CRISPR/Cas9 edited cell line ab264823 (CRISPR/Cas9 edited cell lysate ab257003) lane below 105kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type HeLa and NFKB1 CRISPR/Cas9 edited HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab32360 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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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human prostate carcinoma tissue labelling NFkB p105 / p50 with unpurified ab32360 at a dilution of 1/250.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32360).
数据表及文件
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Datasheet download
Certificate of Compliance
文献 (37)
ab220803 被引用在 37 文献中.
- Zhang T et al. Amyloid precursor protein binds with TNFRSF21 to induce neural inflammation in Alzheimer's Disease. Eur J Pharm Sci 157:105598 (2021). PubMed: 33075465
- Hu C et al. Inhibition of Long Noncoding RNA SNHG15 Ameliorates Hypoxia/Ischemia-Induced Neuronal Damage by Regulating miR-302a-3p/STAT1/NF-?B Axis. Yonsei Med J 62:325-337 (2021). PubMed: 33779086
- Zhang X et al. miR-139-5p protects septic mice with acute lung injury by inhibiting Toll-like receptor 4/Myeloid differentiation factor 88/Nuclear factor-&mac_kgr;B signaling pathway. Clinics (Sao Paulo) 76:e2484 (2021). PubMed: 33681946
- Huang Z et al. C1q/TNF-related protein 9 decreases cardiomyocyte hypoxia/reoxygenation-induced inflammation by inhibiting the TLR4/MyD88/NF-κB signaling pathway. Exp Ther Med 22:1139 (2021). PubMed: 34504585
- Bai S et al. Exosomal circ_DLGAP4 promotes diabetic kidney disease progression by sponging miR-143 and targeting ERBB3/NF-?B/MMP-2 axis. Cell Death Dis 11:1008 (2020). PubMed: 33230102