NFkB p65 Polyclonal Antibody/NFkB p65多克隆抗体

货号：51-0500

规格：100 µg

价格：4809

产品类型：抗体和染料

品牌：Thermo Fisher

抗原：A recombinant protein derived from the carboxyl te

物种：其它

宿主：兔

抗体亚型：IgG

荧光染料：其它

类型:	多抗	同型对照：
浓度：	0.25 mg/mL	用法：	5 µg cells(ChIP)；0.1-1.0 µg/mL(ELISA)；1:100-1:500(ICC)；1:100-1:500(IF)；0.5-2 µg/mL(IHC )；1-3 µg/mL(WB)

靶标信息Nuclear factor kB p65 (NF-kB p65) is encoded by the RELA gene and is present on chromosome 11 in humans. NF-kB P65 is also known as RelA (v-rel avian reticuloendotheliosis viral oncogene homolog A) and belongs to the Rel family of proteins. It is one of the two subunits of NF-kB (Nuclear factor kappa-light-chain-enhancer of activated B cells) that heterodimerizes with the other subunits p50 or p52. However, binding of TNF alpha to its cognate receptor phosphorylates IKK which in turn phosphorylates I kB allowing proteasomal degradation of I kB. Phosphorylation of p65 at Ser 536 located in the carboxy-terminal transactivation domains (TAD) leads to nuclear localization of the transcriptionally active complex and NF-kB mediated transactivation of several downstream genes. It specifically plays a key role in transcription of immunoglobulin k (kappa) gene in mature B-lymphoid cells.

数据：

NFkB p65 Antibody (51-0500) in IFImmunofluorescence analysis of NFkB p65 was done on 70% confluent log phase ME-180 cells. The cells were either mock treated or treated with TNF-alpha (50 ng/mL for 20 min), fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were subsequently labeled with NFkB p65 (Green) Rabbit Polyclonal Antibody (510500) at 1:300 in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature. Nuclei (Blue) were stained with SlowFade® Gold Antifade Mountant DAPI (Product # S36938). F-actin (Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). TNFalpha induced nuclear translocation of NFkB, a downstream target in the TNFR1, TRADD and IKK alpha was observed in control cell line (panels a, e) and not in the TNFR1, TRADD and IKK alpha knockout (KO) cell lines (panels b-d; f-h). The images were captured at 40X magnification. NFkB p65 Antibody (51-0500) in WBWestern blot analysis of NFkB (p65) was performed by loading 30 µg of HeLa (lane1), HEK-293 (lane2), A-431 (lane3), K562 (lane4) Cos-7 (lane5), NIH/3T3 (lane 6), MDCK (lane 7) cell lysate using Novex®NuPAGE®4-12 % Bis-Tris gel (Product # NP0321BOX), XCell SureLock Electrophoresis System (Product # EI0002), Novex® Sharp Pre-Stained Protein Standard (LC5800), and iBlot® Dry Blotting System (IB21001). Proteins were transferred to a nitrocellulose membrane and blocked with 5% skim milk for 1 hour at room temperature. NFkB (p65) was detected at ~ 65 kDa using NFkB (p65) Rabbit Polyclonal Antibody (Product # 51-0500) at 1-3 µg/mL in 5% skim milk at 4°C overnight on a rocking platform. Goat Anti-Rabbit IgG - HRP Secondary Antibody (G21234) at 1:5000 dilution was used and chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

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