| 货号 | 13586S |
| 反应种属 | Human/Mouse/Rat |
| 来源宿主 | Rabbit |
| 应用 | W/IP/IF-IC/F/ChIP |
| 使用方法 | WB(1:1000) IP (1:100) F (1:100) IF-IC (1:200) ChIP (1:50) |
| 供应商 | CST |
| 背景 | Transcription factors of the nuclear factor κB (NF-κB)/Rel family play a pivotal role in inflammatory and immune responses (1,2). There are five family members in mammals: RelA, c-Rel, RelB, NF-κB1 (p105/p50), and NF-κB2 (p100/p52). Both p105 and p100 are proteolytically processed by the proteasome to produce p50 and p52, respectively. Rel proteins bind p50 and p52 to form dimeric complexes that bind DNA and regulate transcription. In unstimulated cells, NF-κB is sequestered in the cytoplasm by IκB inhibitory proteins (3-5). NF-κB-activating agents can induce the phosphorylation of IκB proteins, targeting them for rapid degradation through the ubiquitin-proteasome pathway and releasing NF-κB to enter the nucleus where it regulates gene expression (6-8). NIK and IKKα (IKK1) regulate the phosphorylation and processing of NF-κB2 (p100) to produce p52, which translocates to the nucleus (9-11). |
| 存放说明 | -20C |
| 计算分子量 | 50 Active form. 120 Precursor |
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HeLa cells treated with Human Tumor Necrosis Factor-α (hTNF-α) #8902 (30 ng/ml, 1 hr) and either 10 μl of NF-κB1 p105/p50 (D4P4D) Rabbit mAb or 2 μl of Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human IκBα Promoter Primers #5552, human IL-8 promoter primers, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one. | |
Western blot analysis of extracts from various cell lines and rat spleen using NF-κB1 p105/p50 (D4P4D) Rabbit mAb. | |
Immunoprecipitation of NF-κB1 p105/p50 from Raji cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or NF-κB1 (D4P4D) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot was performed using NF-κB1 p105/p50 (D4P4D) Rabbit mAb. Mouse Anti-rabbit IgG (Light-Chain Specific) (L57A3) mAb #3677 was used as a secondary antibody. | |
Confocal immunofluorescent analysis of C2C12 cells, untreated (left) or treated with Mouse Tumor Necrosis Factor-α (mTNF-α) #5178 (20 ng/ml, 30 min; right), using NF-κB1 p105/p50 (D4P4D) Rabbit mAb (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red). Blue pseudocolor= DRAQ5® #4084 (fluorescent DNA dye). | |
Flow cytometric analysis of C2C12 cells using NF-κB1 p105/p50 (D4P4D) Rabbit mAb (blue) compared to Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (red). Anti-rabbit IgG (H+L), F(ab)2 Fragment (Alexa Fluor® 647 Conjugate) #4414 was used as a secondary antibody. |
