Cell Signaling Technology

Product Pathways - Chromatin Regulation / Epigenetics

SPT4 (D3P2W) Rabbit mAb #64828

No. Size Price
64828S 100 µl ( 10 western blots ) ¥3,250.00 现货查询 购买询价 防伪查询
64828 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human,Mouse,Rat,Monkey, Endogenous 13 Rabbit IgG
IP 1:50
ChIP 1:100

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IP=Immunoprecipitation, ChIP=Chromatin IP,


Species predicted to react based on 100% sequence homology: Hamster, S. cerevisiae,

Specificity / Sensitivity

SPT4 (D3P2W) Rabbit mAb recognizes endogenous levels of total SPT4 protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of human SPT4 protein.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using SPT4 (D3P2W) Rabbit mAb.



Immunoprecipitation of SPT4 from HCT 116 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is SPT4 (D3P2W) Rabbit mAb. Western blot analysis was performed using SPT4 (D3P2W) Rabbit mAb.

Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HCT 116 cells and either 5 μl of SPT4 (D3P2W) 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, SimpleChIP® Human GAPDH Exon 1 Primers #5516, 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.


DRB-sensitivity inducing factor (DSIF), a heterodimer composed of SPT4 and SPT5, is capable of both facilitating and inhibiting RNA polymerase II (RNAPII) activity (1-3). DSIF, together with NELF (Negative Elongation Factor), inhibits RNAPII elongation, resulting in promoter proximal pausing of RNAPII as it awaits additional signaling to resume transcription (4). The release of promoter proximal pausing is signaled through phosphorylation of the RNAPII C-terminal domain (CTD) and NELF by positive transcription elongation factor (P-TEFb) (5). P-TEFb also phosphorylates SPT5 at Thr4 within the evolutionarily conserved heptapeptide repeat motif. This phosphorylation event switches DSIF from a transcriptional repressor to an activator where it becomes a critical factor for transcriptional elongation (6,7).

  1. Wada, T. et al. (1998) Genes Dev 12, 343-56.
  2. Saunders, A. et al. (2006) Nat Rev Mol Cell Biol 7, 557-67.
  3. Gomes, N.P. et al. (2006) Genes Dev 20, 601-12.
  4. Yamaguchi, Y. et al. (1999) Cell 97, 41-51.
  5. Buratowski, S. (2009) Mol Cell 36, 541-6.
  6. Wada, T. et al. (1998) EMBO J 17, 7395-403.
  7. Yamada, T. et al. (2006) Mol Cell 21, 227-37.

Application References

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Companion Products

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

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

XP is a registered trademark of Cell Signaling Technology, Inc.

SimpleChIP is a registered trademark of Cell Signaling Technology, Inc.

Tween is a registered trademark of ICI Americas, Inc.

Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.

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