Anti-SP1 (phospho T453) 抗体 - ChIP Grade (ab59257)

製品の概要

  • 製品名Anti-SP1 (phospho T453) antibody - ChIP Grade
    SP1 一次抗体 製品一覧
  • 製品の詳細
    Rabbit polyclonal to SP1 (phospho T453) - ChIP Grade
  • 特異性ab59257 detects endogenous levels of SP1 only when phosphorylated at threonine 453.
  • アプリケーション適用あり: ELISA, IHC-P, WB, ChIP, ICC/IFmore details
  • 種交差性
    交差種: Mouse, Rat, Human
  • 免疫原

    Synthetic phosphopeptide derived from human SP1 around the phosphorylation site of threonine 453 (I-R-TP-P-T).

  • ポジティブ・コントロール
    • IHC-P: Human brain tissue. WB: A549 cell extracts.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab59257 in the following tested applications.

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

アプリケーション Abreviews 特記事項
ELISA 1/5000.
IHC-P Use at an assay dependent concentration.
WB 1/500 - 1/1000. Detects a band of approximately 90 kDa (predicted molecular weight: 81 kDa).
ChIP Use at an assay dependent concentration. PubMed: 20375015
ICC/IF Use at an assay dependent concentration. PubMed: 21115498

ターゲット情報

  • 機能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.
  • 組織特異性Up-regulated in adenocarcinomas of the stomach (at protein level).
  • 配列類似性Belongs to the Sp1 C2H2-type zinc-finger protein family.
    Contains 3 C2H2-type zinc fingers.
  • 翻訳後修飾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).
  • 細胞内局在Nucleus. Cytoplasm. Nuclear location is governed by glycosylated/phosphorylated states. Insulin promotes nuclear location, while glucagon favors cytoplasmic location.
  • Information by UniProt
  • 参照データベース
  • 別名
    • 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

Anti-SP1 (phospho T453) antibody - ChIP Grade 画像

  • Immunofluorescence analysis of HeLa cells, using ab59257 Antibody. The picture on the right is treated with the synthesized peptide.

  • Wild type MEF cells (2 × 107 cells) were cross-linked with formaldehyde, quenched with glycine, resuspended in SDS lysis buffer (1% SDS, 10 mm EDTA, 50 mm Tris-HCl, pH 8.0, with protease inhibitors and phosphatase inhibitors), sonicated on ice, and centrifuged at 4 °C. Supernatant (400 μl) were diluted to a final volume of 4 ml in a mixture of 9 parts dilution buffer (1% Triton X-100, 150 mm NaCl, 2 mm EDTA, 20 mm Tris-HCl, with protease inhibitors, pH 8.0) and 1 part lysis buffer.

    Mixtures were incubated with 4 μg of anti-p-Sp1 (ab 59257) or anti-Sp1 antibodies with rotating at 4 °C overnight followed with incubation with 100 μl of protein A beads with rotating at 4 °C for 4 h. After gentle centrifugation (2000 rpm), beads were resuspended in 1 ml of wash buffer (1% Triton X-100, 0.1% SDS, 150 mm NaCl, 2 mm EDTA, 20 mm Tris-HCl, with protease inhibitors, pH 8.0) and washed with wash buffer 3 times followed by one wash with a final wash buffer (1% Triton X-100, 0.1% SDS, 500 mm NaCl, 2 mm EDTA, 20 mm Tris-HCl, pH 8.0, with protease inhibitors). The immune complexes were eluted with elution buffer (1% SDS, 100 mm NaHCO3) followed by incubation with proteinase K and RNase A (500 μg/ml each) at 37 °C for 30 min. Reverse cross-links were performed by placing the tubes at 65 °C overnight. Immunoprecipitated DNA was extracted and dissolved in sterile water and Q-PCR was performed.

  • All lanes : Anti-SP1 (phospho T453) antibody - ChIP Grade (ab59257) at 1/500 dilution

    Lane 1 : A549 cell extracts
    Lane 2 : A549 cell extracts with immunising phospho peptide


    Predicted band size : 81 kDa
    Observed band size : 90 kDa (why is the actual band size different from the predicted?)
  • ab59257, at 1/50 dilution, staining SP1 in paraffin embedded human brain tissue by Immunohistochemistry in the absence or presence of the immunising peptide.
  • ab59257 (1:1000) Antibody detects endogenous levels of SP1 only when phosphorylated at Thr453. 

Anti-SP1 (phospho T453) antibody - ChIP Grade (ab59257) 使用論文

This product has been referenced in:
  • Shimoyama S  et al. Dephosphorylation of Sp1 at Ser-59 by protein phosphatase 2A (PP2A) is required for induction of CYP1A1 transcription after treatment with 2,3,7,8-tetrachlorodibenzo-p-dioxin or omeprazole. Biochim Biophys Acta 1839:107-15 (2014). Read more (PubMed: 24382322) »
  • Du Y  et al. NF-?B and enhancer-binding CREB protein scaffolded by CREB-binding protein (CBP)/p300 proteins regulate CD59 protein expression to protect cells from complement attack. J Biol Chem 289:2711-24 (2014). Human . Read more (PubMed: 24338025) »

See all 13 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (HEK293)
Gel Running Conditions Reduced Denaturing
Loading amount 25 µg
Specification HEK293
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
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Abcam user community

Verified customer

投稿 Nov 11 2016

Application Western blot
Sample Mouse Cell lysate - whole cell (Cardiomyocytes)
Gel Running Conditions Reduced Denaturing
Loading amount 25 µg
Specification Cardiomyocytes
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
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投稿 Sep 21 2016

Application Western blot
Loading amount 25 µg
Gel Running Conditions Reduced Denaturing
Sample Human Cell lysate - whole cell (Hep G2)
Specification Hep G2
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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投稿 Dec 29 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Tissue lysate - whole (Melanoma cancer)
Loading amount 30 µg
Specification Melanoma cancer
Treatment 5gy Irradiation
Gel Running Conditions Non-reduced Denaturing (8%)
Blocking step Milk as blocking agent for 1 hour(s) and 30 minute(s) · Concentration: 3% · Temperature: 25°C
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Ms. Seontae Kim

Verified customer

投稿 Jan 14 2013

Thank you for your call today and for letting us know about the trouble with this antibody.

As we discussed, I'm sending a free of charge vial of ab59257 on the order ***, which should arrive on Monday.

Please keep me updated about ...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Rat Cell lysate - nuclear (Intestine)
Loading amount 50 µg
Specification Intestine
Treatment DFO
Gel Running Conditions Non-reduced Denaturing (7.5% gel SDS)
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 25°C
Username

Liwei Xie

Verified customer

投稿 Jun 17 2011

SP1 has a number of glycosylation sites and so this is probably the reason why we see a difference between the expected MW (80-90kDa) (ab59257) and the observed (120kDa) in our product ab37707. We have run ab37707 alongside ab59257. No information wa...

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