NFkB p65 Recombinant Rabbit Monoclonal Antibody (4-2H22L23)/NFkB p65重组兔单克隆抗体（4-2H22L23）

货号：701079

规格：100 µg

价格：4809

产品类型：抗体和染料

品牌：Thermo Fisher

抗原：Recombinant protein corresponding to amino acids 3

物种：人

宿主：兔

抗体亚型：IgG

克隆号：4-2H22L23

荧光染料：其它

类型:	单抗	同型对照：
浓度：	0.5 mg/mL	用法：	1-5 µg x 10^6 cells(ChIP)；1:10-1:50(IHC (P))；1-5 µg/mL(WB)

产品详细信息Intact IgG appears on a non-reducing gel as ~150 kDa band and upon reduction generating a ~25 kDa light chain band and a ~50 kDa heavy chain.Recombinant rabbit monoclonal antibodies are produced using in vitro expression systems. The expression systems are developed by cloning in the specific antibody DNA sequences from immunoreactive rabbits. Then, individual clones are screened to select the best candidates for production. The advantages of using recombinant rabbit monoclonal antibodies include: better specificity and sensitivity, lot-to-lot consistency, animal origin-free formulations, and broader immunoreactivity to diverse targets due to larger rabbit immune repertoire.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 (701079) in IHC (P)Immunohistochemistry analysis of NFKB P65 showing staining in the cytoplasm of paraffin-embedded human breast carcinoma (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a NFKB P65 Recombinant Rabbit Monoclonal Antibody (Product # 701079) diluted in 3% BSA-PBS at a dilution of 1:20 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting. NFkB p65 Antibody (701079) in WBWestern blot analysis was performed on whole cell extracts (30 µg lysate) of HT-1080 (Lane 1), THP-1 (Lane 2), A431 (Lane 3), MCF7 (lane 4) and HEK-293 (lane 5). The blots were probed with Anti-NFkB p65 Recombinant Rabbit Monoclonal Antibody (Product # 701079, 2-3 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). A 60 kDa band corresponding to NFkB p65 was observed across cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 10 % Bis-Tris gel (Product # NP0302BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western blotting Substrate (Product # 32106).

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