Rpb1 CTD Antibody Sampler Kit【科瑞奥博】

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Rpb1 CTD Antibody Sampler Kit

货号： 54020T 基本售价： 4822.0 元规格： -

产品信息

概述
货号	54020T
目标/特异性	Rpb1 NTD (D8L4Y) Rabbit mAb recognizes endogenous levels of total Rpb1 protein at the amino terminal domain (NTD). Phospho-Rpb1 CTD (Ser2) (E1Z3G) Rabbit mAb recognizes endogenous levels of Rpb1 only when the carboxy-terminal domain (CTD) heptapeptide repeat [Tyr1, Ser2, Pro3, Thr4, Ser5, Pro6, Ser7] is phosphorylated at Ser2. This antibody does not cross-react with Rpb1 CTD phosphorylated at Ser5 or Ser7. Phospho-Rpb1 CTD (Ser5) (D9N5I) Rabbit mAb recognizes endogenous levels of Rpb1 only when the carboxy-terminal domain (CTD) heptapeptide repeat [Tyr1, Ser2, Pro3, Thr4, Ser5, Pro6, Ser7] is phosphorylated at Ser5. This antibody does not cross-react with Rpb1 CTD phosphorylated at Ser2 or Ser7. Phospho-Rpb1 CTD (Ser2/Ser5) (D1G3K) Rabbit mAb recognizes endogenous levels of Rpb1 only when the carboxy-terminal domain (CTD) heptapeptide repeat [Tyr1, Ser2, Pro3, Thr4, Ser5, Pro6, Ser7] is dually phosphorylated at Ser2 and Ser5. This antibody does not cross-react with Rpb1 CTD that is singly phosphorylated at Ser2, Ser5, or Ser7. Phospho-Rpb1 CTD (Ser7) (E2B6W) Rabbit mAb recognizes endogenous levels of Rpb1 protein only when the carboxy-terminal domain (CTD) heptapeptide repeat [Tyr1, Ser2, Pro3, Thr4, Ser5, Pro6, Ser7] is phosphorylated at Ser7. This antibody does not cross-react with Rpb1 CTD phosphorylated at Ser2 or Ser5.

性能
供应商	CST
背景	RNA polymerase II (RNAPII) is a large multi-protein complex that functions as a DNA-dependent RNA polymerase, catalyzing the transcription of DNA into RNA using the four ribonucleoside triphosphates as substrates (1). The largest subunit, RNAPII subunit B1 (Rpb1), also known as RNAPII subunit A (POLR2A), contains a unique heptapeptide sequence (Tyr1,Ser2,Pro3,Thr4,Ser5,Pro6,Ser7), which is repeated up to 52 times in the carboxy-terminal domain (CTD) of the protein (1). This CTD heptapeptide repeat is subject to multiple post-translational modifications, which dictate the functional state of the polymerase complex. Phosphorylation of the CTD during the active transcription cycle integrates transcription with chromatin remodeling and nascent RNA processing by regulating the recruitment of chromatin modifying enzymes and RNA processing proteins to the transcribed gene (1). During transcription initiation, RNAPII contains a hypophosphorylated CTD and is recruited to gene promoters through interactions with DNA-bound transcription factors and the Mediator complex (1). The escape of RNAPII from gene promoters requires phosphorylation at Ser5 by CDK7, the catalytic subunit of transcription factor IIH (TFIIH) (2). Phosphorylation at Ser5 mediates the recruitment of RNA capping enzymes, in addition to histone H3 Lys4 methyltransferases, which function to regulate transcription initiation and chromatin structure (3,4). After promoter escape, RNAPII proceeds down the gene to an intrinsic pause site, where it is halted by the negative elongation factors NELF and DSIF (5). At this point, RNAPII is unstable and frequently aborts transcription and dissociates from the gene. Productive transcription elongation requires phosphorylation at Ser2 by CDK9, the catalytic subunit of the positive transcription elongation factor P-TEFb (6). Phosphorylation at Ser2 creates a stable transcription elongation complex and facilitates recruitment of RNA splicing and polyadenylation factors, in addition to histone H3 Lys36 methyltransferases, which function to promote elongation-compatible chromatin (7,8). Ser2/Ser5-phosphorylated RNAPII then transcribes the entire length of the gene to the 3 end, where transcription is terminated. RNAPII dissociates from the DNA and is recycled to the hypophosphorylated form by various CTD phosphatases (1).
运输条件	0.75
存放说明	-20C
参考文献	Brookes, E. and Pombo, A. (2009) EMBO Rep 10, 1213-9. Komarnitsky, P. et al. (2000) Genes Dev 14, 2452-60. Ho, C.K. and Shuman, S. (1999) Mol Cell 3, 405-11. Ng, H.H. et al. (2003) Mol Cell 11, 709-19. Cheng, B. and Price, D.H. (2007) J Biol Chem 282, 21901-12. Marshall, N.F. et al. (1996) J Biol Chem 271, 27176-83. Krogan, N.J. et al. (2003) Mol Cell Biol 23, 4207-18. Proudfoot, N.J. et al. (2002) Cell 108, 501-12. Chapman, R.D. et al. (2007) Science 318, 1780-2. Egloff, S. et al. (2007) Science 318, 1777-9. Egloff, S. et al. (2008) Biochem Soc Trans 36, 590-4. Baillat, D. et al. (2005) Cell 123, 265-76. Akhtar, M.S. et al. (2009) Mol Cell 34, 387-93. Egloff, S. et al. (2010) J Biol Chem 285, 20564-9. Egloff, S. et al. (2012) Mol Cell 45, 111-22.

参考图片

After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO^® is added and emits light during enzyme catalyzed decomposition.

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 10⁶ HeLa cells and either 10 μl of Phospho-Rpb1 CTD (Ser2) (E1Z3G) 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 β-Actin Promoter Primers #13653, human β-Actin intron 1 primers, SimpleChIP^® Human β-Actin 3 UTR Primers #13669, 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.

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 10⁶ HeLa cells and either 10 μl of Phospho-Rpb1 CTD (Ser5) (D9N5I) 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 β-Actin Promoter Primers #13653, human β-Actin intron 1 primers, SimpleChIP^® Human β-Actin 3 UTR Primers #13669, 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.

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 10⁶ HeLa cells and either 10 μl of Phospho-Rpb1 CTD (Ser2/Ser5) (D1G3K) 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 β-Actin Promoter Primers #13653, human Β-Actin intron 1 primers, SimpleChIP^® Human β-Actin 3 UTR Primers #13669, 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 C2C12, H-4-II-E, and COS-7 cells using Phospho-Rpb1 CTD (Ser5) (D9N5I) Rabbit mAb.

Western blot analysis of extracts from C2C12, H-4-II-E, and COS-7 cells using Phospho-Rpb1 CTD (Ser2/Ser5) (D1G3K) Rabbit mAb.

Immunoprecipitation of Rpb1 from HeLa cell extracts using Rabbit (DA1E) mAb IgG XP^® Isotype Control #3900 (lane 2) or Phospho-Rpb1 CTD (Ser2/Ser5) (D1G3K) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using Phospho-Rpb1 CTD (Ser2/Ser5) (D1G3K) Rabbit mAb.

Immunoprecipitation of Rpb1 from HeLa cell extracts using Rabbit (DA1E) mAb IgG XP^® Isotype Control #3900 (lane 2) or Phospho-Rpb1 CTD (Ser5) (D9N5I) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using Phospho-Rpb1 CTD (Ser5) (D9N5I) Rabbit mAb.

Immunoprecipitation of Rpb1 from HeLa cell extracts using Rabbit (DA1E) mAb IgG XP^® Isotype Control #3900 (lane 2) or Phospho-Rpb1 CTD (Ser2) (E1Z3G) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using Phospho-Rpb1 CTD (Ser2) (E1Z3G) Rabbit mAb.

Western blot analysis of extracts from C2C12, H-4-II-E, and COS-7 cells using Phospho-Rpb1 CTD (Ser2) (E1Z3G) Rabbit mAb.

Western blot analysis of extracts from various cell lines using Phospho-Rpb1 CTD (Ser7) (E2B6W) Rabbit mAb.

Immunoprecipitation of Rpb1 from HeLa cell extracts using Rabbit (DA1E) mAb IgG XP^® Isotype Control #3900 (lane 2) or Phospho-Rpb1 CTD (Ser7) (E2B6W) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using Phospho-Rpb1 CTD (Ser7) (E2B6W) Rabbit mAb.

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 10⁶ HeLa cells and either 10 μl of Phospho-Rpb1 CTD (Ser7) (E2B6W) 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 human β-Actin upstream primers, SimpleChIP^®Human β-Actin 3 UTR Primers #13669, human RNU2 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 HeLa, KNRK, and COS-7 cells using Rpb1 NTD (D8L4Y) Rabbit mAb.

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 10⁶ HeLa cells and either 10 μl of Rpb1 NTD (D8L4Y) 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 GAPDH Exon 1 Primers #5516, SimpleChIP^® Human γ-Actin Promoter Primers #5037, SimpleChIP^® Human AFM1 Intron 1 Primers #5098, 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.