Cell Signaling Technology

Product Pathways - Chromatin Regulation / Epigenetics

TH1L (D5G6W) Rabbit mAb #12265

Applications Reactivity Sensitivity MW (kDa) Isotype
W IP ChIP H M R Mk (Hm) (B) (Dg) (Hr) Endogenous 66 Rabbit IgG

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  ChIP=Chromatin IP
Reactivity Key:  H=Human  M=Mouse  R=Rat  Hm=Hamster  Mk=Monkey  B=Bovine  Dg=Dog  Hr=Horse
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

TH1L (D5G6W) Rabbit mAb recognizes endogenous levels of total TH1L protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ala132 of human TH1L protein.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using TH1L (D5G6W) Rabbit mAb.

IP

IP

Immunoprecipitation of TH1L from HCT 116 cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or TH1L (D5G6W) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using TH1L (D5G6W) Rabbit mAb.

Chromatin IP

Chromatin IP

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 and either 5 μl of TH1L (D5G6W) 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 PMAIP1 Intron 1 Primers #12558, human c-Myc 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.


Background

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 awaits 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 C-terminal domain (CTD) within the largest subunit of RNAPII (4,5). 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 (1,6,7). TH1L, together with COBRA-1, are integral subunits that bring WHSC2 and NELF-E together in the NELF complex (1).

  1. Narita, T. et al. (2003) Mol Cell Biol 23, 1863-73.
  2. Yamaguchi, Y. et al. (1999) Cell 97, 41-51.
  3. Nechaev, S. and Adelman, K. (2011) Biochim Biophys Acta 1809, 34-45.
  4. Buratowski, S. (2009) Mol Cell 36, 541-6.
  5. Yamaguchi, Y. et al. (1999) Cell 97, 41-51.
  6. Yamaguchi, Y. et al. (2001) Science 293, 124-7.
  7. Yamaguchi, Y. et al. (2002) Mol Cell Biol 22, 2918-27.

Application References

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For Research Use Only. Not For Use In Diagnostic Procedures.

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