Cell Signaling Technology

Product Pathways - Chromatin Regulation / Epigenetics

SP1 (D4C3) Rabbit mAb #9389

Applications Reactivity Sensitivity MW (kDa) Isotype
W IP IHC-P F ChIP H Mk Endogenous 90 Rabbit IgG

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)  F=Flow Cytometry  ChIP=Chromatin IP
Reactivity Key:  H=Human  Mk=Monkey
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

SP1 (D4C3) Rabbit mAb recognizes endogenous levels of total SP1 protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Pro593 of human SP1 protein.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using SP1 (D4C3) Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human lung carcinoma using SP1 (D4C3) Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human colon carcinoma using SP1 (D4C3) Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human prostate carcinoma using SP1 (D4C3) Rabbit mAb.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of HeLa cells using SP1 (D4C3) Rabbit mAb #9389 (blue) compared to concentration matched Rabbit (DA1E) mAb XP® Isotype Control #3900 (red). Anti-Rabbit IgG (H+L), (F(ab')2 Fragment (Alexa Fluor 488 Conjugate) #4412 was used as a secondary antibody

Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 293 cells and either 5 μl of SP1 (D4C3) 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 using SimpleChIP® Human DHFR Intron 1 Primers #7531, human TIN2 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

Specificity protein 1 (SP1) is a ubiquitously expressed transcription factor belonging to the family of C2H2-type zinc finger containing DNA-binding proteins. SP1 binds GC-rich motifs with high affinity and regulates the expression of numerous mammalian genes (1,2). It interacts with many other transcription factors, such as c-Myc, EGR1, and Stat1, and with basal transcription machinery components. SP1 interacts with chromatin-modifying factors, such as histone deacetylases (HDACs) and p300 in chromatin remodeling. Transcriptional activity and stability of SP1 are regulated by post-translational modification, including phosphorylation, acetylation, ubiquitination, and glycosylation (3). Glycosylation of SP1 following insulin treatment leads to increased nuclear localization, while glucagon treatment increases cytoplasmic SP1 levels (4-6). Investigators have found high levels of SP1 in patients with Alzheimer's disease (7).

  1. Kadonaga, J.T. et al. (1987) Cell 51, 1079-90.
  2. Song, J. et al. (2003) Int J Mol Med 11, 547-53.
  3. Tan, N.Y. and Khachigian, L.M. (2009) Mol Cell Biol 29, 2483-8.
  4. Majumdar, G. et al. (2003) Am J Physiol Endocrinol Metab 285, E584-91.
  5. Majumdar, G. et al. (2006) J Biol Chem 281, 3642-50.
  6. Solomon, S.S. et al. (2008) Life Sci 83, 305-12.
  7. Citron, B.A. et al. (2008) J Neurosci Res 86, 2499-504.

Application References

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Companion Products

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

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