Overview

  • Product name

  • Description

    Rabbit polyclonal to SP1
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IP, IHC-P, ChIP, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
    Predicted to work with: Cow, Rhesus monkey
  • Immunogen

    Recombinant fragment within Human SP1 (internal sequence). The exact sequence is proprietary.
    Database link: P08047

  • Positive control

    • WB: HEK-293T and THP-1 whole cell lysate. ICC/IF: HeLa cells. IHC: HeLa and C2C12 xenografts. IP: THP-1 whole cell lysate. ChIP: HEK-293T chromatin extract.

Properties

Applications

Our Abpromise guarantee covers the use of ab227383 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/500 - 1/20000.
IP 1/100 - 1/500.
IHC-P 1/100 - 1/1000.
ChIP Use at an assay dependent concentration.
ICC/IF 1/100 - 1/1000.

Target

  • Function

    Transcription factor that can activate or repress transcription in response to physiological and pathological stimuli. Binds with high affinity to GC-rich motifs and regulates the expression of a large number of genes involved in a variety of processes such as cell growth, apoptosis, differentiation and immune responses. Highly regulated by post-translational modifications (phosphorylations, sumoylation, proteolytic cleavage, glycosylation and acetylation). Binds also the PDGFR-alpha G-box promoter. May have a role in modulating the cellular response to DNA damage. Implicated in chromatin remodeling. Plays a role in the recruitment of SMARCA4/BRG1 on the c-FOS promoter. Plays an essential role in the regulation of FE65 gene expression. In complex with ATF7IP, maintains telomerase activity in cancer cells by inducing TERT and TERC gene expression.
  • Tissue specificity

    Up-regulated in adenocarcinomas of the stomach (at protein level).
  • Sequence similarities

    Belongs to the Sp1 C2H2-type zinc-finger protein family.
    Contains 3 C2H2-type zinc fingers.
  • Post-translational
    modifications

    Phosphorylated on multiple serine and threonine residues. Phosphorylation is coupled to ubiquitination, sumoylation and proteolytic processing. Phosphorylation on Ser-59 enhances proteolytic cleavage. Phosphorylation on Ser-7 enhances ubiquitination and protein degradation. Hyperphosphorylation on Ser-101 in response to DNA damage has no effect on transcriptional activity. MAPK1/MAPK3-mediated phosphorylation on Thr-453 and Thr-739 enhances VEGF transcription but, represses FGF2-triggered PDGFR-alpha transcription. Also implicated in the repression of RECK by ERBB2. Hyperphosphorylated on Thr-278 and Thr-739 during mitosis by MAPK8 shielding SP1 from degradation by the ubiquitin-dependent pathway. Phosphorylated in the zinc-finger domain by calmodulin-activated PKCzeta. Phosphorylation on Ser-641 by PKCzeta is critical for TSA-activated LHR gene expression through release of its repressor, p107. Phosphorylation on Thr-668, Ser-670 and Thr-681 is stimulated by angiotensin II via the AT1 receptor inducing increased binding to the PDGF-D promoter. This phosphorylation is increased in injured artey wall. Ser-59 and Thr-681 can both be dephosphorylated by PP2A during cell-cycle interphase. Dephosphorylation on Ser-59 leads to increased chromatin association during interphase and increases the transcriptional activity. On insulin stimulation, sequentially glycosylated and phosphorylated on several C-terminal serine and threonine residues.
    Acetylated. Acetylation/deacetylation events affect transcriptional activity. Deacetylation leads to an increase in the expression the 12(s)-lipooxygenase gene though recruitment of p300 to the promoter.
    Ubiquitinated. Ubiquitination occurs on the C-terminal proteolytically-cleaved peptide and is triggered by phosphorylation.
    Sumoylated by SUMO1. Sumoylation modulates proteolytic cleavage of the N-terminal repressor domain. Sumoylation levels are attenuated during tumorigenesis. Phosphorylation mediates SP1 desumoylation.
    Proteolytic cleavage in the N-terminal repressor domain is prevented by sumoylation. The C-terminal cleaved product is susceptible to degradation.
    O-glycosylated; contains at least 8 N-acetylglucosamine side chains. Levels are controlled by insulin and the SP1 phosphorylation states. Insulin-mediated O-glycosylation locates SP1 to the nucleus, where it is sequentially deglycosylated and phosphorylated. O-glycosylation affects transcriptional activity through disrupting the interaction with a number of transcription factors including ELF1 and NFYA. Also inhibits interaction with the HIV1 promoter. Inhibited by peroxisomome proliferator receptor gamma (PPARgamma).
  • Cellular localization

    Nucleus. Cytoplasm. Nuclear location is governed by glycosylated/phosphorylated states. Insulin promotes nuclear location, while glucagon favors cytoplasmic location.
  • Information by UniProt
  • Database links

  • Alternative names

    • SP 1 antibody
    • SP1 antibody
    • Sp1 transcription factor antibody
    • SP1_HUMAN antibody
    • Specificity protein 1 antibody
    • Transcription factor Sp1 antibody
    • TSFP 1 antibody
    • TSFP1 antibody
    see all

Images

  • All lanes : Anti-SP1 antibody (ab227383) at 1/10000 dilution

    Lane 1 : Non-transfected HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate
    Lane 2 : SP1 shRNA transfected HEK-293T whole cell lysate

    Lysates/proteins at 50 µg per lane.

    Secondary
    All lanes : HRP-conjugated anti-rabbit IgG

    Developed using the ECL technique.

    Performed under reducing conditions.
  • HeLa (human epithelial cell line from cervix adenocarcinoma) cells stained for SP1 (green) using ab227383 at 1/500 dilution in ICC/IF. Cells were fixed in 4% paraformaldehyde at RT for 15 minutes. Red: phalloidin, a cytoskeleton marker, diluted at 1/200.

  • ChIP was performed with HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen)chromatin extract and 5 μg of either normal rabbit IgG or anti-SP1 antibody. The precipitated DNA was detected by PCR with primer set targeting to MGARP promoter.

  • SP1 was immunoprecipitated from THP-1 (human monocytic leukemia cell line) whole cell lysate with ab227383. Western blot was performed from the immunoprecipitate using ab227383. Anti-rabbit IgG was used as secondary antibody.

    Lane 1: THP-1 whole cell lysate (Input).

    Lane 2: Rabbit  IgG instead of ab227383 in THP-1 whole cell lysate.

    Lane 3: ab227383 IP in THP-1 whole cell lysate.

  • Paraffin-embedded HeLa (human epithelial cell line from cervix adenocarcinoma) xenograft stained for SP1 using ab227383 at 1/500 dilution in immunohistochemical analysis.

  • Paraffin-embedded C2C12 (mouse myoblast cell line) xenograft stained for SP1 using ab227383 at 1/500 dilution in immunohistochemical analysis.

  • Anti-SP1 antibody (ab227383) at 1/2000 dilution + THP-1 (human monocytic leukemia cell line) whole cell lysate at 30 µg

    Secondary
    HRP-conjugated anti-rabbit IgG

    Developed using the ECL technique.

    Performed under reducing conditions.


    7.5% SDS-PAGE

References

This product has been referenced in:

  • Dong H  et al. Activation of LncRNA TINCR by H3K27 acetylation promotes Trastuzumab resistance and epithelial-mesenchymal transition by targeting MicroRNA-125b in breast Cancer. Mol Cancer 18:3 (2019). Read more (PubMed: 30621694) »
  • Yu S  et al. SP1-induced lncRNA TINCR overexpression contributes to colorectal cancer progression by sponging miR-7-5p. Aging (Albany NY) 11:1389-1403 (2019). Read more (PubMed: 30853664) »
See all 5 Publications for this product

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