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#54020 Rpb1 CTD Antibody Sampler Kit

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2019年3月20日11時40分 現在
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キット内容 容量 用途 種交差性 ホモロジー† 検出分子量 アイソタイプ
Phospho-Rpb1 CTD (Ser2) (E1Z3G) Rabbit mAb #13499 20 µl WB, IP, ChIP, ChIP-seq H, M, R, Mk Hm, Dm, X, Z, B, Pg, Sc, Ce 250 Rabbit IgG
Phospho-Rpb1 CTD (Ser5) (D9N5I) Rabbit mAb #13523 20 µl WB, IP, ChIP, ChIP-seq H, M, R, Mk Hm, Dm, X, B, Pg, Sc, Ce 250 Rabbit IgG
Phospho-Rpb1 CTD (Ser2/Ser5) (D1G3K) Rabbit mAb #13546 20 µl WB, IP, ChIP, ChIP-seq H, M, R, Mk Hm, Dm, X, Z, B, Pg, Sc, Ce 250 Rabbit IgG
Phospho-Rpb1 CTD (Ser7) (E2B6W) Rabbit mAb #13780 20 µl WB, IP, ChIP H, M, R, Mk 250 Rabbit IgG
Rpb1 NTD (D8L4Y) Rabbit mAb #14958 20 µl WB, ChIP, ChIP-seq H, M, R, Mk Hm, B 250 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody #7074 100 µl WB Goat

†Species predicted to react based on 100% sequence homology.

Applications Key: W=Western Blotting, IP=Immunoprecipitation, ChIP=Chromatin IP, ChIP-seq=Chromatin IP-seq
Reactivity Key: H=Human, M=Mouse, R=Rat, Mk=Monkey

貯法
-20℃
社内データ

Western Blotting

Western Blotting

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.

Western Blotting

Western Blotting

Western blot analysis of extracts from C2C12, H-4-II-E, and COS-7 cells using Phospho-Rpb1 CTD (Ser5) (D9N5I) Rabbit mAb.

Western Blotting

Western Blotting

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


Western Blotting

Western Blotting

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

Western Blotting

Western Blotting

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

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa, KNRK, and COS-7 cells using Rpb1 NTD (D8L4Y) Rabbit mAb.


Chromatin IP-seq

Chromatin IP-seq

Chromatin immunoprecipitations were performed with cross-linked chromatin from Hela cells and Rpb1 NTD (D8L4Y) Rabbit mAb, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. The figure shows binding across the ZNF740 gene on chromosome 12. For additional ChIP-seq tracks, please download the product data sheet.

Chromatin IP-seq

Chromatin IP-seq

Chromatin immunoprecipitations were performed with cross-linked chromatin from Hela cells and either Phospho-Rpb1 CTD (Ser2) (E1Z3G) Rabbit mAb or Rpb1 NTD (D8L4Y) Rabbit mAb 314958, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. The figure shows binding across the ZNF740 gene on chromosome 12. For additional ChIP-seq tracks, please download the product data sheet.

Chromatin IP-seq

Chromatin IP-seq

Chromatin immunoprecipitations were performed with cross-linked chromatin from Hela cells and either Phospho-Rpb1 CTD (Ser5) (D9N5I) Rabbit mAb or Rpb1 NTD (D8L4Y) Rabbit mAb #14958, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. The figure shows binding across the ZNF740 gene on chromosome 12. For additional ChIP-seq tracks, please download the product data sheet.


Chromatin IP-seq

Chromatin IP-seq

Chromatin immunoprecipitations were performed with cross-linked chromatin from Hela cells and either Phospho-Rpb1 CTD (Ser2/Ser5) (D1G3K) Rabbit mAb or Rpb1 NTD (D8L4Y) Rabbit mAb #14958, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. The figure shows binding across the ZNF740 gene on chromosome 12. For additional ChIP-seq tracks, please download the product data sheet.

Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells and either Phospho-Rpb1 CTD (Ser2) (E1Z3G) Rabbit mAb or 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 IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells and either Phospho-Rpb1 CTD (Ser5) (D9N5I) Rabbit mAb or 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 IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells and either Phospho-Rpb1 CTD (Ser2/Ser5) (D1G3K) Rabbit mAb or 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.

IP

IP

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.

IP

IP

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.


IP

IP

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.

IP

IP

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 IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells and either Phospho-Rpb1 CTD (Ser7) (E2B6W) Rabbit mAb or 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.


Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells and either Rpb1 NTD (D8L4Y) Rabbit mAb or 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.

バックグラウンド

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).

In addition to Ser2/Ser5 phosphorylation, Ser7 of the CTD heptapeptide repeat is also phosphorylated during the active transcription cycle. Phosphorylation at Ser7 is required for efficient transcription of small nuclear (sn) RNA genes (9,10). snRNA genes, which are neither spliced nor poly-adenylated, are structurally different from protein-coding genes. Instead of a poly(A) signal found in protein-coding RNAs, snRNAs contain a conserved 3'-box RNA processing element, which is recognized by the Integrator snRNA 3' end processing complex (11,12). Phosphorylation at Ser7 by CDK7 during the early stages of transcription facilitates recruitment of RPAP2, which dephosphorylates Ser5, creating a dual Ser2/Ser7 phosphorylation mark that facilitates recruitment of the Integrator complex and efficient processing of nascent snRNA transcripts (13-15).

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13499   Phospho-Rpb1 CTD (Ser2) (E1Z3G) Rabbit mAb
13523   Phospho-Rpb1 CTD (Ser5) (D9N5I) Rabbit mAb
13546   Phospho-Rpb1 CTD (Ser2/Ser5) (D1G3K) Rabbit mAb
13780   Phospho-Rpb1 CTD (Ser7) (E2B6W) Rabbit mAb
14958   Rpb1 NTD (D8L4Y) Rabbit mAb
6883   SignalFire™ ECL Reagent
7074   Anti-rabbit IgG, HRP-linked Antibody
7075   Anti-biotin, HRP-linked Antibody
7727   Biotinylated Protein Ladder Detection Pack

Illumina is a registered trademark of Illumina, Inc.
Tween is a registered trademark of ICI Americas, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.
SimpleChIP is a registered trademark of Cell Signaling Technology, Inc.
LumiGLO is a registered trademark of Kirkegaard & Perry Laboratories.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

本製品は試験研究用です。

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