Overview

  • Product nameAnti-SP1 antibody - ChIP GradeSee all SP1 primary antibodies ...
  • Description
    Rabbit polyclonal to SP1 - ChIP Grade
  • Tested applicationsIP, WB, IHC-P, ChIP more details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
    Predicted to work with: Rabbit, Goat, Horse, Chicken, Cow, Dog, Pig, Xenopus laevis, Chimpanzee, Zebrafish, Rhesus monkey, Gorilla, Orangutan, Xenopus tropicalis
  • Immunogen

    Synthetic peptide (Unfortunately the amino acid sequence is considered to be commercially sensitive). The peptide sequence is within the region between aa750 and the C-terminus (aa785) of human SP1 using the numbering given in entry XP_028606.4 (GeneID 666).

  • Positive control
    • IHC: Breast Carcinoma, Lung Adenocarcinoma, Ovarian Carcinoma.

Properties

Applications

Our Abpromise guarantee covers the use of ab13370 in the following tested applications.

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

Application Notes
IP Use at 1-4 µg/mg of lysate.
WB 1/2500 - 1/10000. Detects a band of approximately 90 kDa (predicted molecular weight: 90 kDa).
IHC-P Use at an assay dependent concentration.
ChIP 1/10.

Target

  • FunctionTranscription 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 specificityUp-regulated in adenocarcinomas of the stomach (at protein level).
  • Sequence similaritiesBelongs 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 localizationNucleus. Cytoplasm. Nuclear location is governed by glycosylated/phosphorylated states. Insulin promotes nuclear location, while glucagon favors cytoplasmic location.
  • Target information above from: UniProt accession P08047 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    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

Anti-SP1 antibody - ChIP Grade images

  • ChIP analysis using ab13370 binding SP1 in Human primary macrophage. Cells were cross-linked for 15 minutes with formaldehyde. Samples were incubated with primary antibody (1/500) for 12 hours at 4°C. Protein binding was detected using real-time PCR.

     

    See Abreview

  • Detection of Human SP1 by Western Blot and Immunoprecipitation.  Samples: A. Whole cell lysate (50 mcg for E; 10 mcg for T) from HEK 293T cells that were mock transfected (E) or transfected with a Sp1 expression construct (T).  B. Whole cell lysate (700 mcg) from HEK 293T cells.  Antibody: A. Affinity purified rabbit anti-Sp1 antibody  (ab13370) used at the indicated concentrations.  B. SP1 was immunoprecipitated using affinity purified rabbit anti-SP1 antibodies (lane 2: ab13405, lane 3: ab13370; lane 1: SP-1 antibody, details unknown) using each antibody at 3.75 mcg/mg lysate.  Subsequently, ab13370 was used at 0.04 mcg/ml for WB. Detection: Chemiluminescence with an exposure time of 10 seconds (A) or 1 min (B).

  • ab13370 (2µg/ml) staining SP1 in human colon using an automated system (DAKO Autostainer Plus). Using this protocol there is nuclear staining.
    Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer EDTA pH 9.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.

References for Anti-SP1 antibody - ChIP Grade (ab13370)

This product has been referenced in:
  • Sharma A  et al. The role of SIRT6 protein in aging and reprogramming of human induced pluripotent stem cells. J Biol Chem 288:18439-47 (2013). ChIP . Read more (PubMed: 23653361) »
  • Lee SJ  et al. 5-Lipoxygenase plays a pivotal role in endothelial adhesion of monocytes via an increased expression of Mac-1. Cardiovasc Res 99:724-33 (2013). Read more (PubMed: 23720274) »

See all 12 Publications for this product

Product Wall

DISCOUNT CODE: 4XXXXX Expiration date: 28-04-2012 I am very pleased to hear you would like to accept our offer and test ab13370 in ChIP on Rat samples. This code will give you 1 free PRIMARY ANTIBODY before the expiration date. To redeem this offer, pl...

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Application ChIP
Detection step Real-time PCR
Sample Human Cell lysate - whole cell (primary human macrophages)
Specification primary human macrophages
Type Cross-linking (X-ChIP)
Duration of cross-linking step: 15 minute(s) and 0 second(s)
Specification of the cross-linking agent: formaldehyde
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Submitted Jan 17 2014

Application Western blot
Sample Mouse Cell lysate - whole cell (Intestine tissue)
Loading amount 50 µg
Specification Intestine tissue
Gel Running Conditions Reduced Denaturing (7.5% gel SDS)
Blocking step (agent) for 1 hour(s) and 0 minute(s) · Concentration: 5µg/mL · Temperature: 25°C
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Liwei Xie

Verified customer

Submitted Jul 05 2012

DISCOUNT CODE: ***
Expiration date: November 3rd, 2012

I am very pleased to hear you would like to accept our offer and test ab13370 in mouse. This code will give you: 1 freeprimary antibodybefore the expiration date. To redeem this offe...

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Application Western blot
Sample Rat Cell lysate - nuclear (Intestine tissue)
Loading amount 50 µg
Specification Intestine tissue
Gel Running Conditions Reduced Denaturing (7.5% gel SDS)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Liwei Xie

Verified customer

Submitted Jul 03 2012

Application ChIP
Sample Rat Cell lysate - nuclear (rat PC12 cell)
Specification rat PC12 cell
Type Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: formaldehyde (final conc: 1% )
Detection step Semiquantitative PCR
Positive control In the DNA electrophoretogram, lane 4 is another anti-SP1 Ab (millipore, Cat. 17-601) that reacts with rat.
Negative control In the DNA electrophoretogram, lane2 is anti-rabbit IgG.
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Submitted May 02 2012

DISCOUNT CODE: ***
Expiration date: August 4th, 2012

I am very pleased to hear you would like to accept our offer and test ab13370 in mouse or rat. This code will give you 1 free primary antibody before the expiration date. To redeem thi...

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Application Immunoprecipitation
Sample Human Cell lysate - nuclear (K562)
Total protein in input 120 µg
Specification K562
Immuno-precipitation step Protein G
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Submitted Feb 15 2012

Thanks for your call today and for letting us know about the trouble with ab13370. I'm sorry to hear that the results were so poor with this antibody. As we discussed, I am sending a free of charge replacement with ab77441 on the order ***, which shoul...

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DISCOUNT CODE: 10*** Expiration date: February 8, 2012 I am very pleased to hear you would like to accept our offer and test ab13370 in IHC-Fr. This code will give you 1 free primary antibody before the expiration date. To redeem this offer, pl...

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"