Объем | 100 мкг |
Синонимы | Transcription factor p65 (Nuclear factor NF-kappa-B p65 subunit) (Nuclear factor of kappa light polypeptide gene enhancer in B-cells 3), RELA, NFKB3 |
Клональность | Polyclonal Antibody |
Организм | Human |
uniprot | Q04206 |
Иммуноген | Recombinant Human Transcription factor p65 protein (1-210AA) |
Источник | Rabbit |
Видовая специфичность | Human, Mouse |
Применение | ELISA, WB, IHC, IF, ChIP, Recommended dilution: WB:1:500-1:5000, IHC:1:500-1:1000, IF:1:50-1:500 |
Примечание | NF-kappa-B is a pleiotropic transcription factor present in almost all cell types and is the endpoint of a series of signal transduction events that are initiated by a vast array of stimuli related to many biological processes 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. |
Клональность1 | Polyclonal |
Изотип | IgG |
Коньюгат | Non-conjugated |
Буффер | Preservative: 0.03% Proclin 300<br />Constituents: 50% Glycerol, 0.01M PBS, PH 7.4 |
Форма | Liquid |
Хранение | Upon receipt, store at -20°C or -80°C. Avoid repeated freeze. |
Метод очистки | >95%, Protein G purified |
Области исследований | Epigenetics and Nuclear Signaling, Neuroscience, Cancer, Cardiovascular, Cell biology, Metabolism, Microbiology, Signal transduction |
Ссылка на страницу на сайте производителя | ссылка |
Western Blot
Positive WB detected in: 293 whole cell lysate, NIH/3T3 whole cell lysate
All lanes: RELA antibody at 3µg/ml
Secondary
Goat polyclonal to rabbit IgG at 1/50000 dilution
Predicted band size: 61, 59, 60 kDa
Observed band size: 61 kDa
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IHC image of CSB-PA03987A0Rb diluted at 1:600 and staining in paraffin-embedded human placenta tissue performed on a Leica BondTM system. After dewaxing and hydration, antigen retrieval was mediated by high pressure in a citrate buffer (pH 6.0). Section was blocked with 10% normal goat serum 30min at RT. Then primary antibody (1% BSA) was incubated at 4°C overnight. The primary is detected by a biotinylated secondary antibody and visualized using an HRP conjugated SP system.
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IHC image of CSB-PA03987A0Rb diluted at 1:600 and staining in paraffin-embedded human breast cancer performed on a Leica BondTM system. After dewaxing and hydration, antigen retrieval was mediated by high pressure in a citrate buffer (pH 6.0). Section was blocked with 10% normal goat serum 30min at RT. Then primary antibody (1% BSA) was incubated at 4°C overnight. The primary is detected by a biotinylated secondary antibody and visualized using an HRP conjugated SP system.
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Immunofluorescence staining of PC-3 cells with CSB-PA03987A0Rb at 1:200, counter-stained with DAPI. The cells were fixed in 4% formaldehyde, permeabilized using 0.2% Triton X-100 and blocked in 10% normal Goat Serum. The cells were then incubated with the antibody overnight at 4°C. The secondary antibody was Alexa Fluor 488-congugated AffiniPure Goat Anti-Rabbit IgG(H+L).
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Chromatin Immunoprecipitation Hela (1.2*106) were cross-linked with formaldehyde, sonicated, and immunoprecipitated with 4µg anti-NF-kB p65 or a control normal rabbit IgG. The resulting ChIP DNA was quantified using real-time PCR with primers (CSB-PP03987HU) against the IkBα promoter.
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IHC image of CSB-PA03987A0Rb diluted at 1:200 and staining in paraffin-embedded human lung tissue performed on a Leica BondTM system. After dewaxing and hydration, antigen retrieval was mediated by high pressure in a citrate buffer (pH 6.0). Section was blocked with 10% normal goat serum 30min at RT. Then primary antibody (1% BSA) was incubated at 4°C overnight. The primary is detected by a biotinylated secondary antibody and visualized using an HRP conjugated SP system.
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Immunofluorescence staining of HepG2 cells with CSB-PA03987A0Rb at 1:66, counter-stained with DAPI. The cells were fixed in 4% formaldehyde, permeabilized using 0.2% Triton X-100 and blocked in 10% normal Goat Serum. The cells were then incubated with the antibody overnight at 4°C. The secondary antibody was Alexa Fluor 488-congugated AffiniPure Goat Anti-Rabbit IgG(H+L).
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