NFkB p50 Monoclonal Antibody (5D10D11)/NFkB p50单克隆抗体（5D10D11）

货号：MA5-15870

规格：100 µg

价格：5468

产品类型：抗体和染料

品牌：Thermo Fisher

抗原：Purified recombinant fragment of human NFKB1 expre

物种：人

宿主：小鼠

抗体亚型：其它

克隆号：5D10D11

荧光染料：其它

类型:	单抗	同型对照：
浓度：	1 mg/mL	用法：	2 µg/1x10^6 cells(ChIP)；1:50-1:200(Flow)；1:100-1:200(ICC)；1:100-1:200(IF)；1:50-1:200(IHC (P))；1:1000(WB)

产品详细信息MA5-15870 targets NFKB1 in FACS, IF, IHC, ChIP, and WB applications and shows reactivity with Human samples.靶标信息NFkB is a nuclear transcription factor activated by various extra and intracellular stimuli such as cytokines, UV radiation, stress, in injury, and by bacterial and viral products. NFkB is involved in regulation of various cellular events including cell growth, differentiation, proliferation, apoptosis and inflammation. NFKB1 (p50) is a 50KDa functional sub-unit of NF-kB and a member of Rel protein family which is synthesized as a p105 precursor protein and consists of an N-terminal conserved RHD-region containing nuclear localization signal, DNA-binding and dimerization domains. NFKB1 (p50) forms homodimers or heterodimerizes with p65, forming the functional NFkB factor. Further, NFKB1 (p50) directs the nuclear translocation of NF-kB and is instrumental in its DNA-binding. The NFkB complex is expressed in most cell types, and is primarily found in the cytoplasm in an inactive state in association with I-kappa-B. Phosphorylation of I-kappa-B releases the complex, allowing translocation to the nucleus and modification of gene expression. Phosphorylation of the p50 subunit at the Ser-337 residue has been shown to significantly increase DNA binding efficiency of the NFkB protein complex. The pathological role of NFkB has been suggested in AIDS, hematogenic cancer cell metastasis and rheumatoid arthritis.

数据：

NFkB p50 Antibody (MA5-15870) in IFImmunofluorescent analysis of NFkB1 (green) in A549 cells either left untreated or treated 20 ng/mL TNF-alpha for 20 hours. The cells were fixed with 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 15 minutes, and blocked with 3% BSA for 30 minutes at room temperature. Cells were stained with a NFkB1mouse monoclonal antibody (Product # MA5-15870) at a concentration of 5 µg/mL in blocking buffer for 1 hour at room temperature, and then incubated with a Goat anti-Mouse IgG (H+L) Secondary Antibody, Alexa Fluor Plus 488 conjugate (Product # A32731) at a dilution of 1:500 for at least 30 minutes at a room temperature in the dark (green). Nuclei (blue) were stained with Hoechst 33342 (Product # 62249). Images were taken on a Thermo Scientific ToxInsight Instrument at 20X magnification. NFkB p50 Antibody (MA5-15870) in ChIPMultiplex microplate Matrix ChIP has been described in detail (http://www.ncbi.nlm.nih.gov/pubmed/25959381). Briefly HTC116 cells were starved followed by addition of serum and samples of cells were cross-linked with formaldehyde after the time points indicated on the x-axis (0, 5 and 30 min). Chromatin was sheared using a Bioruptor and ChIP assays were performed using protein A-coated 96-well polypropylene microplates with 1uL/100uL well volume of NF kappa B monoclonal antibody (Product # MA5-15870). Quantitative real-time PCRs were performed in quadruplicate using 1 to 2uL of DNA with primers to -15kb downstream of Egr1 and exon 1 of Erg1. PCR calibration curves were generated for each primer pair from a dilution series of total human genomic DNA. The PCR primer efficiency curve was fit to cycle threshold (Ct) versus log [genomic DNA concentration] by using an r2 best fit. DNA concentration values for each ChIP and input DNA sample were calculated from their respective average Ct values. Final results are expressed as fraction of input DNA. Schematic representations of Erg1 loci is shown where boxes represent exons (black boxes = translated regions, white boxes = untranslated regions), the zigzag line represents an intron, and the straight line represents upstream sequence. Regions amplified by the primers are represented by black bars. Data courtesy of Dr. Karol Bomsztyk’s laboratory

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