货号 | 14894S |
反应种属 | Human/Mouse/Rat/Monkey |
来源宿主 | Rabbit |
应用 | W/IP/ChIP |
使用方法 | WB(1:1000) IP (1:50) ChIP (1:50) |
供应商 | CST |
背景 | Negative Elongation Factor (NELF) consists of four subunits: WHSC2 (NELF-A), COBRA-1 (NELF-B), TH1L (NELF-C/D), and NELF-E (1). NELF, together with DRB-sensitivity inducing factor (DSIF), inhibits RNA Polymerase II (RNAPII) elongation resulting in RNAPII promoter proximal pausing, where it waits additional signaling to resume transcription (2,3). The release of RNAPII from promoter proximal pausing is a critical regulatory point during transcription and is signaled by positive transcription elongation factor (p-TEF-b) phosphorylation of both NELF and the carboxy-terminal domain (CTD) within the largest subunit of RNAPII (3,4). WHSC2 is thought to connect the NELF complex to RNAPII machinery, while NELF-E contains an RNA binding motif that is necessary for NELF function (1,5,6). TH1L, together with COBRA-1, are integral subunits that bring WHSC2 and NELF-E together in the NELF complex (1). |
存放说明 | -20C |
计算分子量 | 65 |
Western blot analysis of extracts from various cell lines using COBRA1 (D6K9A) Rabbit mAb. | |
Immunoprecipitation of COBRA1 from MCF7 cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or COBRA1 (D6K9A) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using COBRA1 (D6K9A) Rabbit mAb. | |
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 MCF7 cells grown in phenol red free medium and 5% charcoal stripped FBS for 4 d then treated with β-estradiol (10 nM) for 45 min and either 10 μl of COBRA1 (D6K9A) 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 c-Myc Intron 1 Primers #14905, human PMAIP1 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. |