Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-CGBP antibody [EPR19199] - BSA and Azide free (ab242431)

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Overview

  • Product name

    Anti-CGBP antibody [EPR19199] - BSA and Azide free
    See all CGBP primary antibodies

  • Description

    Rabbit monoclonal [EPR19199] to CGBP - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: ChIP, IHC-P, WB, ICC/IF, IP, Flow Cytmore 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

    • 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

    Ab242431 is the carrier-free version of ab198977. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    ab242431 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    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.

Properties

Applications

Our Abpromise guarantee covers the use of ab242431 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.
IHC-P Use at an assay dependent concentration. 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
WB Use at an assay dependent concentration. Detects a band of approximately 76 kDa (predicted molecular weight: 76 kDa).
ICC/IF Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.

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

    see all

  • 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

  • Western blot - Anti-CGBP antibody [EPR19199] - BSA and Azide free (ab242431)
    Western blot - Anti-CGBP antibody [EPR19199] - BSA and Azide free (ab242431)
    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).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab198977).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CGBP antibody [EPR19199] - BSA and Azide free (ab242431)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CGBP antibody [EPR19199] - BSA and Azide free (ab242431)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab198977).

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

  • Immunocytochemistry/ Immunofluorescence - Anti-CGBP antibody [EPR19199] - BSA and Azide free (ab242431)
    Immunocytochemistry/ Immunofluorescence - Anti-CGBP antibody [EPR19199] - BSA and Azide free (ab242431)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab198977).

  • Flow Cytometry - Anti-CGBP antibody [EPR19199] - BSA and Azide free (ab242431)
    Flow Cytometry - Anti-CGBP antibody [EPR19199] - BSA and Azide free (ab242431)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab198977).

  • ChIP - Anti-CGBP antibody [EPR19199] - BSA and Azide free (ab242431)
    ChIP - Anti-CGBP antibody [EPR19199] - BSA and Azide free (ab242431)

    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
    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab198977).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CGBP antibody [EPR19199] - BSA and Azide free (ab242431)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CGBP antibody [EPR19199] - BSA and Azide free (ab242431)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab198977).

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

  • Immunocytochemistry/ Immunofluorescence - Anti-CGBP antibody [EPR19199] - BSA and Azide free (ab242431)
    Immunocytochemistry/ Immunofluorescence - Anti-CGBP antibody [EPR19199] - BSA and Azide free (ab242431)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab198977).

  • Flow Cytometry - Anti-CGBP antibody [EPR19199] - BSA and Azide free (ab242431)
    Flow Cytometry - Anti-CGBP antibody [EPR19199] - BSA and Azide free (ab242431)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab198977).

  • Immunoprecipitation - Anti-CGBP antibody [EPR19199] - BSA and Azide free (ab242431)
    Immunoprecipitation - Anti-CGBP antibody [EPR19199] - BSA and Azide free (ab242431)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab198977)

References

ab242431 has not yet been referenced specifically in any publications.

Publishing research using ab242431? Please let us know so that we can cite the reference in this datasheet.

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