Overview

  • Product name
    Anti-SP1 antibody - ChIP Grade
    See all SP1 primary antibodies
  • Description
    Rabbit polyclonal to SP1 - ChIP Grade
  • Host species
    Rabbit
  • Tested applications
    Suitable for: IP, WB, IHC-P, ChIPmore details
  • Species reactivity
    Reacts with: Rat, Human
    Predicted to work with: Rabbit, Goat, Horse, Chicken, Cow, Dog, Pig, Xenopus laevis, Chimpanzee, Rhesus monkey, Gorilla, Orangutan, Zebra finch, 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 Abreviews 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

  • 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

  • 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

  • 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.
  • SP1 was immunoprecipitated from HeLa cells whole cell lysate with ab13370. Anti-SP1 antibody - ChIP Grade (ab13370) used at 6µg per reaction. SP1 was also immunoprecipitated by a previous lot of this antibody. For blotting immunoprecipitated SP1, ab13370 was used at 1µg/ml. Chemiluminescence with an exposure time of 10 seconds.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue sections labeling SP1 with ab13370 at 1/5000 dilution. DAB detection.

  • 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).

References

This product has been referenced in:
  • Hashemi P  et al. Compounds producing an effective combinatorial regimen for disruption of HIV-1 latency. EMBO Mol Med 10:160-174 (2018). WB ; Human . Read more (PubMed: 29246970) »
  • Li S  et al. Long non-coding RNA metastasis-associated lung adenocarcinoma transcript 1 promotes lung adenocarcinoma by directly interacting with specificity protein 1. Cancer Sci 109:1346-1356 (2018). Read more (PubMed: 29575609) »
See all 40 Publications for this product

Customer reviews and Q&As

1-10 of 18 Abreviews or Q&A

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

Abcam user community

Verified customer

Submitted Jan 17 2014

Answer

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, please submit an Abreview for ChIP on Rat samples] and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews. Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code. Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research. The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.

Read More
Application
ChIP
Sample
Mouse Cell lysate - whole cell (Breast)
Specification
Breast
Detection step
Real-time PCR
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: Formalin

Abcam user community

Verified customer

Submitted Feb 25 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
ChIP
Sample
Human Cell lysate - whole cell (Breast)
Specification
Breast
Detection step
Real-time PCR
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: Formalin

Abcam user community

Verified customer

Submitted Feb 25 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (Breast)
Gel Running Conditions
Reduced Denaturing (12.5%)
Loading amount
1e+006 cells
Specification
Breast
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C

Abcam user community

Verified customer

Submitted Feb 10 2016

Application
Western blot
Sample
Mouse Cell lysate - whole cell (Breast)
Gel Running Conditions
Reduced Denaturing (12.5)
Loading amount
1e+006 cells
Specification
Breast
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C

Abcam user community

Verified customer

Submitted Feb 10 2016

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

Liwei Xie

Verified customer

Submitted Jul 05 2012

Question
Answer

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 offer, please submit an Abreview formouse and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. Please remember that submission of the Abreview is sufficient for the discount code to become active.
For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews.

Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code.

Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.

The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
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

Liwei Xie

Verified customer

Submitted Jul 03 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
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.

Abcam user community

Verified customer

Submitted May 02 2012

1-10 of 18 Abreviews or Q&A

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