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54020
Rpb1 CTD Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

Rpb1 CTD Antibody Sampler Kit #54020

Citations (4)
Simple Western™ analysis of lysates (1 mg/ml) from C2C12 cells using Phospho-Rpb1 CTD (Ser2) (E1Z3G) Rabbit mAb #13499. The virtual lane view (left) shows the target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 66-440 kDa.
Simple Western™ analysis of lysates (0.1 mg/mL) from H-4-II-E cells using Phospho-Rpb1 CTD (Ser5) (D9N5I) Rabbit mAb #13523. The virtual lane view (left) shows the target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 66-440 kDa separation module.
Simple Western™ analysis of lysates (0.1 mg/mL) from H-4-II-E cells using Phospho-Rpb1 CTD (Ser2/Ser5) (D1G3K) Rabbit mAb #13546. The virtual lane view (left) shows the target band (as indicated) at 1:50 and 1:250 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:50 (blue line) and 1:250 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 66-440 kDa separation module.
Simple WesternTM analysis of lysates (0.1 mg/mL) from Hela cells using Phospho-Rpb1 CTD (Ser7) (E2B6W) Rabbit mAb #13780. The virtual lane view (left) shows the target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the JessTM Simple Western instrument from ProteinSimple, a BioTechne brand, using the 44-660 kDa separation module.
Western blot analysis of extracts from C2C12, H-4-II-E, and COS-7 cells using Phospho-Rpb1 CTD (Ser2) (E1Z3G) Rabbit mAb.
CUT&Tag was performed with HeLa cells and Phospho-Rpb1 CTD (Ser2) (E1Z3G) Rabbit mAb, using CUT&Tag Assay Kit #77552. DNA library was prepared using CUT&Tag Dual Index Primers and PCR Master Mix for Illumina Systems #47415. The figure shows binding across ACTB gene.
Western blot analysis of extracts from C2C12, H-4-II-E, and COS-7 cells using Phospho-Rpb1 CTD (Ser5) (D9N5I) Rabbit mAb.
CUT&RUN was performed with HeLa cells and Phospho-Rpb1 CTD (Ser5) (D9N5I) Rabbit mAb, using CUT&RUN Assay Kit #86652. DNA Library was prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figures show binding across chromosome 7 (upper), including ACTB (lower), a known target gene of Rpb1 (see additional figure containing CUT&RUN-qPCR data).
Western blot analysis of extracts from C2C12, H-4-II-E, and COS-7 cells using Phospho-Rpb1 CTD (Ser2/Ser5) (D1G3K) Rabbit mAb.
CUT&Tag was performed with HeLa cells and Phospho-Rpb1 CTD (Ser2/Ser5) (D1G3K) Rabbit mAb, using CUT&Tag Assay Kit #77552. DNA library was prepared using CUT&Tag Dual Index Primers and PCR Master Mix for Illumina Systems #47415. The figure shows binding across ACTB, a known target gene of Rpb1 (see our ChIP-qPCR figure).
CUT&RUN was performed with HeLa cells and Phospho-Rpb1 CTD (Ser2/Ser5) (D1G3K) Rabbit mAb, using CUT&RUN Assay Kit #86652. DNA Library was prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across ACTB gene.
Western blot analysis of extracts from various cell lines using Phospho-Rpb1 CTD (Ser7) (E2B6W) Rabbit mAb.

Chromatin immunoprecipitations were performed with cross-linked chromatin from Hela cells and either Phospho-Rpb1 CTD (Ser7) (E2B6W) 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 datasheet.
Western blot analysis of extracts from HeLa, KNRK, and COS-7 cells using Rpb1 NTD (D8L4Y) Rabbit mAb.
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.
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.
CUT&Tag was performed with HeLa cells and Phospho-Rpb1 CTD (Ser2) (E1Z3G) Rabbit mAb, using CUT&Tag Assay Kit #77552. DNA library was prepared using CUT&Tag Dual Index Primers and PCR Master Mix for Illumina Systems #47415. The figures show binding across chromosome 7 (upper), including ACTB gene (lower).
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.
CUT&RUN was performed with HeLa cells and Phospho-Rpb1 CTD (Ser5) (D9N5I) Rabbit mAb, using CUT&RUN Assay Kit #86652. DNA Library was prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across ACTB, a known target gene of Rpb1 (see additional figure containing CUT&RUN-qPCR data).
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.
CUT&Tag was performed with HeLa cells and Phospho-Rpb1 CTD (Ser2/Ser5) (D1G3K) Rabbit mAb, using CUT&Tag Assay Kit #77552. DNA library was prepared using CUT&Tag Dual Index Primers and PCR Master Mix for Illumina Systems #47415. The figures show binding across chromosome 7 (upper), including ACTB (lower), a known target gene of Rpb1 (see our ChIP-qPCR figure).
CUT&RUN was performed with HeLa cells and Phospho-Rpb1 CTD (Ser2/Ser5) (D1G3K) Rabbit mAb, using CUT&RUN Assay Kit #86652. DNA Library was prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figures show binding across chromosome 7 (upper), including ACTB gene (lower).
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 Hela cells and either Phospho-Rpb1 CTD (Ser7) (E2B6W) 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 chromosome 7 (upper), including ACTB (medium), a known target gene of Phospho-Rpb1 CTD (Ser7) (see additional figure containing ChIP-qPCR data), and ZNF740 gene on chromosome 12 (lower).
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 DNA Library Prep Kit for Illumina Systems (ChIP-seq, CUT&RUN) #56795. The figure shows binding across the ZNF740 gene on chromosome 12. For additional ChIP-seq tracks, please download the product datasheet.
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 DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across the ZNF740 gene on chromosome 12.
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 DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across the ZNF740 gene on chromosome 12. For additional ChIP-seq tracks, please download the product datasheet.
CUT&RUN was performed with HeLa cells and either Phospho-Rpb1 CTD (Ser5) (D9N5I) Rabbit mAb or Rabbit (DA1E) mAb IgG XP® Isotype Control (CUT&RUN) #66362, using CUT&RUN Assay Kit #86652. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human β-Actin Promoter Primers #13653, human β-Actin intron 1 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 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 DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across the ZNF740 gene on chromosome 12. For additional ChIP-seq tracks, please download the product datasheet.
CUT&RUN was performed with HeLa cells and either Phospho-Rpb1 CTD (Ser2/Ser5) (D1G3K) Rabbit mAb or Rabbit (DA1E) mAb IgG XP® Isotype Control (CUT&RUN) #66362, using CUT&RUN Assay Kit #86652. The enriched DNA was quantified by real-time PCR using human CKS1B exon 1 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 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 DNA Library Prep Kit for Illumina Systems (ChIP-seq, CUT&RUN) #56795. The figure shows binding across chromosome 7 (upper), including ACTB (medium), a known target gene of Rpb1-NTD (see additional figure containing ChIP-qPCR data), and ZNF740 gene on chromosome 12 (lower).
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 DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across chromosome 7 (upper), including ACTB (medium), a known target gene of Phospho-Rpb1 CTD (Ser2) (see additional figure containing ChIP-qPCR data), and ZNF740 gene on chromosome 12 (lower).
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 DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across chromosome 7 (upper), including ACTB (medium), a known target gene of Phospho-Rpb1 CTD (Ser5) (see additional figure containing ChIP-qPCR data), and ZNF740 gene on chromosome 12 (lower).
Chromatin immunoprecipitations were performed with cross-linked chromatin from Hela cells and either Phospho-Rpb1 CTD (Ser2/Ser5) (D1G3K) Rabbit mAb or of Rpb1 NTD (D8L4Y) Rabbit mAb #14958, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across chromosome 7 (upper), including ACTB (medium), a known target gene of Phospho-Rpb1 CTD (Ser2/Ser5) (see additional figure containing ChIP-qPCR data), and ZNF740 gene on chromosome 12 (lower).
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 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.
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 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 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.
CUT&RUN was performed with HCT 116 cells and Phospho-Rpb1 CTD (Ser2) (E1Z3G) Rabbit mAb, using CUT&RUN Assay Kit #86652. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across ACTB, a known target gene of Rpb1 (see additional figure containing CUT&RUN-qPCR data).
CUT&RUN was performed with HCT 116 cells and Phospho-Rpb1 CTD (Ser2) (E1Z3G) Rabbit mAb, using CUT&RUN Assay Kit #86652. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figures show binding across chromosome 7 (upper), including ACTB (lower), a known target gene of Rpb1 (see additional figure containing CUT&RUN-qPCR data).
CUT&RUN was performed with HeLa cells and either Phospho-Rpb1 CTD (Ser2) (E1Z3G) Rabbit mAb or Rabbit (DA1E) mAb IgG XP® Isotype Control (CUT&RUN) #66362, using CUT&RUN Assay Kit #86652. The enriched DNA was quantified by real-time PCR using human β-actin intron 1 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.
To Purchase # 54020
Cat. # Size Qty. Price
54020T
1 Kit  (5 x 20 microliters)

Product Includes Quantity Applications Reactivity MW(kDa) Isotype
Phospho-Rpb1 CTD (Ser2) (E1Z3G) Rabbit mAb 13499 20 µl
  • WB
  • IP
  • ChIP
  • C&R
  • C&T
H M R Mk 250 Rabbit IgG
Phospho-Rpb1 CTD (Ser5) (D9N5I) Rabbit mAb 13523 20 µl
  • WB
  • IP
  • ChIP
  • C&R
H M R Mk 250 Rabbit IgG
Phospho-Rpb1 CTD (Ser2/Ser5) (D1G3K) Rabbit mAb 13546 20 µl
  • WB
  • IP
  • ChIP
  • C&R
  • C&T
H M R Mk 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
H M R Mk 250 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

Product Description

The Rpb1 CTD Antibody Sampler Kit provides an economical means of evaluating total Rpb1 NTD levels as well as Rpb1 CTD phosphorylated and specific sites. The kit includes enough primary antibodies to perform two western blot experiments per primary antibody.

Specificity / Sensitivity

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.

Source / Purification

Monoclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser2 of the human Rpb1 CTD heptapeptide repeat, Ser5 of the human Rpb1 CTD heptapeptide repeat, Ser2/5 of the human Rpb1 CTD heptapeptide repeat, Ser7 of the human Rpb1 CTD heptapeptide repeat, and a synthetic peptide corresponding to residues surrounding Glu613 of human Rpb1 protein.

Background

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).

Pathways

Explore pathways related to this product.

Limited Uses

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