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遺伝子ノックアウト細胞株 検証済リコンビナントRabMAb

Anti-SP1 抗体 [EPR6662(B)] (ab124804)

Reviews (1) Submit a question References (30)
ChIC/CUT&RUN sequencing - Anti-SP1 antibody [EPR6662(B)] (ab124804)
  • Western blot - Anti-SP1 antibody [EPR6662(B)] (ab124804)
  • Flow Cytometry (Intracellular) - Anti-SP1 antibody [EPR6662(B)] (ab124804)
  • Immunocytochemistry/ Immunofluorescence - Anti-SP1 antibody [EPR6662(B)] (ab124804)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SP1 antibody [EPR6662(B)] (ab124804)
  • Western blot - Anti-SP1 antibody [EPR6662(B)] (ab124804)
  • Western blot - Anti-SP1 antibody [EPR6662(B)] (ab124804)
  • Western blot - Anti-SP1 antibody [EPR6662(B)] (ab124804)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SP1 antibody [EPR6662(B)] (ab124804)
  • Western blot - Anti-SP1 antibody [EPR6662(B)] (ab124804)
  • OI-RD Scanning - Anti-SP1 antibody [EPR6662(B)] (ab124804)
  • Anti-SP1 antibody [EPR6662(B)] (ab124804)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR6662(B)] to SP1
  • Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF, ChIC/CUT&RUN-seq
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 555 Alexa Fluor® 568 Alexa Fluor® 594 Alexa Fluor® 647 Alexa Fluor® 750 Alkaline Phosphatase APC Carrier Free HRP PE

製品の概要

  • 製品名

    Anti-SP1 antibody [EPR6662(B)]
    SP1 一次抗体 製品一覧

  • 製品の詳細

    Rabbit monoclonal [EPR6662(B)] to SP1

  • 由来種

    Rabbit

  • アプリケーション

    適用あり: Flow Cyt (Intra), WB, IHC-P, ICC/IF, ChIC/CUT&RUN-seqmore details
    適用なし: IP

  • 種交差性

    交差種: Human
    非交差種: Mouse, Rat

  • 免疫原

    Synthetic peptide within Human SP1 aa 550-650. The exact sequence is proprietary.

  • ポジティブ･コントロール

    • WB: Ramos, HAP1, HeLa K562 and Raji cell lysates. IHC-P: human gastric carcinoma, Human colon tissue. ICC/IF: HeLa cells. Flow Cyt (intra): HeLa cells. ChIC/CUT&RUN-Seq: HeLa cells.

  • 特記事項

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

製品の特性

アプリケーション

The Abpromise guarantee

Abpromise保証は、 次のテスト済みアプリケーションにおけるab124804の使用に適用されます

アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。

アプリケーション Abreviews 特記事項
Flow Cyt (Intra)
1/100.
WB (1)
1/1000 - 1/10000. Detects a band of approximately 90 kDa (predicted molecular weight: 81 kDa).
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

(Heat to 98&degC, allow to cool for 10-20 minutes)

See IHC antigen retrieval protocols
ICC/IF
1/100 - 1/250.
ChIC/CUT&RUN-seq
Use at an assay dependent concentration.

5µg
特記事項
Flow Cyt (Intra)
1/100.
WB
1/1000 - 1/10000. Detects a band of approximately 90 kDa (predicted molecular weight: 81 kDa).
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

(Heat to 98&degC, allow to cool for 10-20 minutes)

See IHC antigen retrieval protocols
ICC/IF
1/100 - 1/250.
ChIC/CUT&RUN-seq
Use at an assay dependent concentration.

5µg
追加情報
Is unsuitable for IP.

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

    画像

    • ChIC/CUT&RUN sequencing - Anti-SP1 antibody [EPR6662(B)] (ab124804)
      ChIC/CUT&RUN sequencing - Anti-SP1 antibody [EPR6662(B)] (ab124804)

      ChIC/CUT&RUN was performed using a pAG-MNAse at a final concentration of 700 ng/mL. 2.5X10^5 of Human wild-type HeLa cell line (ab255928) or Human SP1 knockout HeLa cell line (ab265519) were used along with 5µg of ab124804 [EPR6662(B)]. Assay Quality Control was conducted using 5µg Anti-CTCF (ab188408) on the same cell lines. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads.  The negative IgG control ab172730 is also shown.

      Additional screenshots of mapped reads can be downloaded here.

      The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.

    • Western blot - Anti-SP1 antibody [EPR6662(B)] (ab124804)
      Western blot - Anti-SP1 antibody [EPR6662(B)] (ab124804)
      All lanes : Anti-SP1 antibody [EPR6662(B)] (ab124804) at 1/5000 dilution

      Lane 1 : Wild-type HeLa cell lysate
      Lane 2 : SP1 knockout HeLa cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 81 kDa
      Observed band size: 100 kDa why is the actual band size different from the predicted?



      Lanes 1-2: Merged signal (red and green). Green - ab124804 observed at 100 kDa. Red - loading control ab8245 observed at 37 kDa.

       ab124804 Anti-SP1 antibody [EPR6662(B)] was shown to specifically react with SP1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265519 (knockout cell lysate ab257698) was used. Wild-type and SP1 knockout samples were subjected to SDS-PAGE. ab124804 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 5000 and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

       

       

    • Flow Cytometry (Intracellular) - Anti-SP1 antibody [EPR6662(B)] (ab124804)
      Flow Cytometry (Intracellular) - Anti-SP1 antibody [EPR6662(B)] (ab124804)

      Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling SP1 with Purified ab124804 at 1/100 dilution (10 µg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

       

    • Immunocytochemistry/ Immunofluorescence - Anti-SP1 antibody [EPR6662(B)] (ab124804)
      Immunocytochemistry/ Immunofluorescence - Anti-SP1 antibody [EPR6662(B)] (ab124804)
      Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling SP1 with Purified ab124804 at 1:200 dilution (4 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with None. Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SP1 antibody [EPR6662(B)] (ab124804)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SP1 antibody [EPR6662(B)] (ab124804)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human gastric carcinoma tissue sections labeling SP1 with Purified ab124804 at 1:2000 dilution (0.41 µg/ml). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). Purified)ImmunoHistoProbe one step HRP Polymer (ready to use) was used for detection. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
    • Western blot - Anti-SP1 antibody [EPR6662(B)] (ab124804)
      Western blot - Anti-SP1 antibody [EPR6662(B)] (ab124804)
      All lanes : Anti-SP1 antibody [EPR6662(B)] (ab124804) at 1/5000 dilution (Purified)

      Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates at 20 µg


      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 81 kDa
      Observed band size: 100 kDa why is the actual band size different from the predicted?



      We are unsure how to define the extra bands.

    • Western blot - Anti-SP1 antibody [EPR6662(B)] (ab124804)
      Western blot - Anti-SP1 antibody [EPR6662(B)] (ab124804)
      Anti-SP1 antibody [EPR6662(B)] (ab124804) at 1/5000 dilution (Purified) + Ramos (Human Burkitt's lymphoma B lymphocyte) whole cell lysates at 15 µg

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 81 kDa
      Observed band size: 100 kDa why is the actual band size different from the predicted?



      We are unsure how to define the extra bands.

    • Western blot - Anti-SP1 antibody [EPR6662(B)] (ab124804)
      Western blot - Anti-SP1 antibody [EPR6662(B)] (ab124804)
      All lanes : Anti-SP1 antibody [EPR6662(B)] (ab124804) at 1/1000 dilution

      Lane 1 : Wild-type HAP1 cell lysate
      Lane 2 : SP1 knockout HAP1 cell lysate
      Lane 3 : HeLa cell lysate
      Lane 4 : K562 cell lysate

      Lysates/proteins at 20 µg per lane.

      Predicted band size: 81 kDa



      Lanes 1 - 4: Merged signal (red and green). Green - ab124804 observed at 100 kDa. Red - loading control, ab8245, observed at 37 kDa.

      Ab16032 detected the expected band for SP1 in wild-type HAP1 cells along with additional cross-reactive bands. The band was not seen in SP1 knockout HAP1 cells. Wild-type and SP1 knockout samples were subjected to SDS-PAGE. ab124804 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/2000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

       

       

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SP1 antibody [EPR6662(B)] (ab124804)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SP1 antibody [EPR6662(B)] (ab124804)

      ab124804, at 1/100 dilution staining SP1 in paraffin-embedded Human colon tissue, by Immunohistochemistry.

      Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    • Western blot - Anti-SP1 antibody [EPR6662(B)] (ab124804)
      Western blot - Anti-SP1 antibody [EPR6662(B)] (ab124804)
      All lanes : Anti-SP1 antibody [EPR6662(B)] (ab124804)

      Lane 1 : Wild-type HAP1 cell lysate
      Lane 2 : SP1 knockout HAP1 cell lysate
      Lane 3 : HeLa cell lysate
      Lane 4 : K562 cell lysate

      Lysates/proteins at 20 µg per lane.

      Predicted band size: 81 kDa



      Lanes 1 - 4: Merged signal (red and green). Green - ab124804 observed at 100 kDa. Red - loading control, ab8245, observed at 37 kDa.

      This western blot image is a comparison between ab124804 and a competitor's top cited rabbit polyclonal antibody.

    • OI-RD Scanning - Anti-SP1 antibody [EPR6662(B)] (ab124804)
      OI-RD Scanning - Anti-SP1 antibody [EPR6662(B)] (ab124804)
      Equilibrium disassociation constant (KD)
      Learn more about KD

      Click here to learn more about KD
    • Anti-SP1 antibody [EPR6662(B)] (ab124804)
      Anti-SP1 antibody [EPR6662(B)] (ab124804)

    参考文献 (30)

    ab124804 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

    ab124804 は 30 報の論文で使用されています。

    • Xu X  et al. Sp1 promotes tumour progression by remodelling the mitochondrial network in cervical cancer. J Transl Med 21:307 (2023). PubMed: 37147632
    • Shi M  et al. UVRAG Promotes Tumor Progression through Regulating SP1 in Colorectal Cancer. Cancers (Basel) 15:N/A (2023). PubMed: 37173968
    • Wu F  et al. TGF-βRII regulates glucose metabolism in oral cancer-associated fibroblasts via promoting PKM2 nuclear translocation. Cell Death Discov 8:3 (2022). PubMed: 35013150
    • Sheikh A  et al. Enterotoxigenic Escherichia coli heat-labile toxin drives enteropathic changes in small intestinal epithelia. Nat Commun 13:6886 (2022). PubMed: 36371425
    • Yuen JG  et al. Development of 5-FU-modified tumor suppressor microRNAs as a platform for novel microRNA-based cancer therapeutics. Mol Ther 30:3450-3461 (2022). PubMed: 35933584
    View all Publications for this product

    レビューと Q&A

    Show All レビュー Q&A
    レビューを送る 質問を送る

    Western blot abreview for Anti-SP1 antibody [EPR6662(B)]

     Excellent
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - nuclear (OCI-LY1)
    Gel Running Conditions
    Reduced Denaturing (10)
    Loading amount
    20 µg
    Specification
    OCI-LY1
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Dr. Xiao Qing Lu

    Verified customer

    投稿 Jan 27 2016

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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