Overview

  • Product name

    Anti-CGBP antibody [EPR19199] - ChIP Grade
    See all CGBP primary antibodies
  • Description

    Rabbit monoclonal [EPR19199] to CGBP - ChIP Grade
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ChIP, WB, IHC-P, ICC/IF, Flow Cyt, IPmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Mouse CGBP aa 100-200. The exact sequence is proprietary.
    Database link: Q9CWW7

  • Positive control

    • WB: 293T, K562, Jurkat, MOLT-4, HeLa, RAW 264,7, PC-12, L-929, NIH/3T3, L6 and F9 whole cell lysates; Human fetal heart and fetal kidney lysates; Mouse brain, mouse heart, rat brain and rat spleen lysates. IHC-P: Mouse cerebrum and rat cerebellum tissues. ICC/IF: Hela and NIH/3T3 cells. Flow Cyt: 293T and NIH/3T3 cells. IP: HeLa whole cell lysate.
  • General notes

     

     

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

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab198977 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
ChIP Use at an assay dependent concentration.
WB 1/2000. Detects a band of approximately 76 kDa (predicted molecular weight: 76 kDa).
IHC-P 1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

IHC is recommended for rat and mouse only

ICC/IF 1/250.
Flow Cyt 1/150.
IP 1/40.

Target

  • Function

    Transcriptional activator that exhibits a unique DNA binding specificity for [AC]CpG[AC] unmethylated CpG motifs.
  • Tissue specificity

    Ubiquitous.
  • Sequence similarities

    Contains 1 CXXC-type zinc finger.
    Contains 1 PHD-type zinc finger.
  • Domain

    The acidic domain carries the potential to activate transcription.
  • Post-translational
    modifications

    May be regulated by proteolysis.
  • Cellular localization

    Nucleus speckle. Associated with euchromatin. During mitosis, excluded from condensed chromosomes.
  • Information by UniProt
  • Database links

  • Alternative names

    • 2410002I16Rik antibody
    • 5830420C16Rik antibody
    • AI426635 antibody
    • CFP 1 antibody
    • CFP1 antibody
    • CpG binding protein antibody
    • CpG-binding protein antibody
    • CXXC 1 antibody
    • CXXC finger 1 antibody
    • CXXC finger 1 PHD domain antibody
    • CXXC finger protein 1 antibody
    • CXXC type zinc finger protein 1 antibody
    • CXXC-type zinc finger protein 1 antibody
    • CXXC1 antibody
    • CXXC1_HUMAN antibody
    • DNA binding protein with PHD finger and CXXC domain antibody
    • hCGBP antibody
    • HsT2645 antibody
    • PCCX 1 antibody
    • PCCX1 antibody
    • PHD finger and CXXC domain containing protein 1 antibody
    • PHD finger and CXXC domain-containing protein 1 antibody
    • PHF 18 antibody
    • PHF18 antibody
    • Protein containing CXXC domain 1 antibody
    • SPP 1 antibody
    • SPP1 antibody
    • ZCGPC 1 antibody
    • ZCGPC1 antibody
    • Zinc finger, CpG binding type containing 1 antibody
    see all

Images

  • All lanes : Anti-CGBP antibody [EPR19199] - ChIP Grade (ab198977) at 1/20000 dilution

    Lane 1 : 293T (Human epithelial cell line from embryonic kidney) whole cell lysate
    Lane 2 : K562 (Human chronic myelogenous leukemia cell line from bone marrow) whole cell lysate
    Lane 3 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
    Lane 4 : MOLT-4 (Human lymphoblastic leukemia cell line) whole cell lysate
    Lane 5 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 76 kDa
    Observed band size: 76 kDa



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1/2/3: 30 seconds; Lane 4: 3 minutes; Lane 5: 10 seconds.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling CGBP with ab198977 at 1/250 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

    Confocal image showing nuclear staining on HeLa cell line.

    The nuclear counterstain is DAPI (blue).

    Tubulin is detected with Anti-alpha Tubulin-Loading Control (ab7291)  at 1/1000 dilution and Goat Anti-Mouse IgG (AlexaFluor®594) preadsorbed (ab150120) at 1/1000 dilution (red).

    The negative controls are as follows:-

    -ve control 1: ab198977 at 1/250 dilution followed by ab150120  at 1/1000 dilution.

    -ve control 2: ab7291  at 1/1000 dilution followed by ab150077  at 1/1000 dilution.

  • Immunohistochemical analysis of paraffin-embedded Rat cerebellum tissue labeling CGBP with ab198977 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. 

    Nuclear staining on rat cerebellum tissue is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Chromatin was prepared from HeLa cells according to the Abcam Dual X-ChIP protocol*. Cells were fixed with EGS for 30 minutes, then formaldehyde for 10 minutes.
    The ChIP was performed with 25 µg of chromatin, 5 µg of ab198977 (red), and 20 µl of Protein A/G sepharose beads. 5 µg of rabbit normal IgG was added to the beads control (gray). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).
    Primers and probes are located in the first kb of the transcribed region.
    *http://www.abcam.com/resources?keywords=X%20ChIP%20protocol
  • All lanes : Anti-CGBP antibody [EPR19199] - ChIP Grade (ab198977) at 1/2000 dilution

    Lane 1 : Human fetal heart lysate
    Lane 2 : Human fetal kidney lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

    Predicted band size: 76 kDa
    Observed band size: 76 kDa


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-CGBP antibody [EPR19199] - ChIP Grade (ab198977) at 1/2000 dilution

    Lane 1 : Mouse brain lysate
    Lane 2 : Mouse heart lysate
    Lane 3 : Rat brain lysate
    Lane 4 : Rat spleen lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 76 kDa
    Observed band size: 76 kDa



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1: 10 seconds; Lane 2/3/4: 3 minutes.

  • Lanes 1-4 : Anti-CGBP antibody [EPR19199] - ChIP Grade (ab198977) at 1/2000 dilution
    Lanes 5-6 : Anti-CGBP antibody [EPR19199] - ChIP Grade (ab198977) at 1/10000 dilution

    Lane 1 : RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate
    Lane 2 : PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate
    Lane 3 : L-929 (Mouse connective tissue fibroblast cell line) whole cell lysate
    Lane 4 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate
    Lane 5 : L6 (Rat skeletal muscle cell line) whole cell lysate
    Lane 6 : F9 (Mouse embryonic testicular cancer cell line) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 76 kDa
    Observed band size: 76 kDa



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1/2: 10 seconds; Lane 3/4/6: 8 seconds; Lane 5: 3 minutes.

  • All lanes : Anti-CGBP antibody [EPR19199] - ChIP Grade (ab198977) at 1/500 dilution

    Lane 1 : WT HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 2 : CFP1(CGBP) KO HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate

    Secondary
    All lanes : Goat anti-Rabbit IgG (H+L), HRP at 1/5000 dilution

    Predicted band size: 76 kDa
    Observed band size: 76 kDa


    Exposure time: 15 seconds


    Blocking buffer: 5% NFDM/TBST.

    Dilution buffer: 1% BSA/TBST.

     

    This data is from our collaborator Hengyu-Fan's lab (Life Sciences Institute Zhejiang University).

  • Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling CGBP with ab198977 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Nuclear staining on mouse cerebrum tissue is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (Mouse embryonic fibroblast cell line) cells labeling CGBP with ab198977 at 1/250 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

    Confocal image showing nuclear staining on NIH/3T3 cell line.

    The nuclear counterstain is DAPI (blue).

    Tubulin is detected Anti-alpha Tubulin antibody - Loading Control (ab7291) at  1/1000 dilution and Goat Anti-Mouse IgG  (AlexaFluor®594) preadsorbed (ab150120) at 1/1000 dilution (red).

    The negative controls are as follows:-

    -ve control 1: ab198977 at 1/250 dilution followed by ab150120 at 1/1000 dilution.

    -ve control 2: ab7291  at 1/1000 dilution followed by ab150077  at 1/1000 dilution.

  • Flow cytometric analysis of 4% paraformaldehyde-fixed 293T (Human epithelial cell line from embryonic kidney) cells labeling CGBP with ab198977 at 1/150 dilution (red) compared with a Rabbit IgG,monoclonal [EPR25A] - Isotype control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti Rabbit IgG (FITC) at 1/500 dilution was used as the secondary antibody.

  • Flow cytometric analysis of 4% paraformaldehyde-fixed NIH/3T3 cells labeling CGBP with ab198977 at 1/150 dilution (red) compared with a Rabbit IgG,monoclonal-Isotype control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti Rabbit IgG (FITC) at a dilution of 1/500 was used as the secondary antibody.

  • CGBP was immunoprecipitated from 1mg of HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab198977 at 1/40 dilution.

    Western blot was performed from the immunoprecipitate using ab198977 at 1/1000 dilution.

    VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

    Lane 1: HeLa whole cell lysate, 10µg (Input).

    Lane 2: ab198977 IP in HeLa whole cell lysate.

    Lane 3: Rabbit IgG,monoclonal [EPR25A] -Isotype Control (ab172730) instead of ab198977 in HeLa whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 1 second.

References

This product has been referenced in:

  • Tian H  et al. CXXC1 is not essential for normal DNA double-strand break formation and meiotic recombination in mouse. PLoS Genet 14:e1007657 (2018). Read more (PubMed: 30365547) »
  • Yu C  et al. CFP1 Regulates Histone H3K4 Trimethylation and Developmental Potential in Mouse Oocytes. Cell Rep 20:1161-1172 (2017). WB, ICC . Read more (PubMed: 28768200) »
See all 3 Publications for this product

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