Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-CGBP antibody [EPR19199] - ChIP Grade (ab198977)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR19199] to CGBP - ChIP Grade
  • Suitable for: ChIP, WB, IHC-P, ICC/IF, Flow Cyt, IP
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

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 & Species

    Application Species
    ChIP
    Human
    Flow Cyt
    Mouse
    Human
    ICC/IF
    Mouse
    Human
    IHC-P
    Mouse
    Rat
    IP
    Human
    WB
    Mouse
    Rat
    Human
    See all applications and species data

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • 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

     

     

    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.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

Images

  • Western blot - Anti-CGBP antibody [EPR19199] - ChIP Grade (ab198977)
    Western blot - Anti-CGBP antibody [EPR19199] - ChIP Grade (ab198977)
    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.

  • Immunocytochemistry/ Immunofluorescence - Anti-CGBP antibody [EPR19199] - ChIP Grade (ab198977)
    Immunocytochemistry/ Immunofluorescence - Anti-CGBP antibody [EPR19199] - ChIP Grade (ab198977)

    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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CGBP antibody [EPR19199] - ChIP Grade (ab198977)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CGBP antibody [EPR19199] - ChIP Grade (ab198977)

    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.

  • ChIP - Anti-CGBP antibody [EPR19199] - ChIP Grade (ab198977)
    ChIP - Anti-CGBP antibody [EPR19199] - ChIP Grade (ab198977)
    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
  • Western blot - Anti-CGBP antibody [EPR19199] - ChIP Grade (ab198977)
    Western blot - Anti-CGBP antibody [EPR19199] - ChIP Grade (ab198977)
    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.

  • Western blot - Anti-CGBP antibody [EPR19199] - ChIP Grade (ab198977)
    Western blot - Anti-CGBP antibody [EPR19199] - ChIP Grade (ab198977)
    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.

  • Western blot - Anti-CGBP antibody [EPR19199] - ChIP Grade (ab198977)
    Western blot - Anti-CGBP antibody [EPR19199] - ChIP Grade (ab198977)
    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.

  • Western blot - Anti-CGBP antibody [EPR19199] - ChIP Grade (ab198977)
    Western blot - Anti-CGBP antibody [EPR19199] - ChIP Grade (ab198977)
    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).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CGBP antibody [EPR19199] - ChIP Grade (ab198977)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CGBP antibody [EPR19199] - ChIP Grade (ab198977)

    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.

  • Immunocytochemistry/ Immunofluorescence - Anti-CGBP antibody [EPR19199] - ChIP Grade (ab198977)
    Immunocytochemistry/ Immunofluorescence - Anti-CGBP antibody [EPR19199] - ChIP Grade (ab198977)

    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 Cytometry - Anti-CGBP antibody [EPR19199] - ChIP Grade (ab198977)
    Flow Cytometry - Anti-CGBP antibody [EPR19199] - ChIP Grade (ab198977)

    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 Cytometry - Anti-CGBP antibody [EPR19199] - ChIP Grade (ab198977)
    Flow Cytometry - Anti-CGBP antibody [EPR19199] - ChIP Grade (ab198977)

    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.

  • Immunoprecipitation - Anti-CGBP antibody [EPR19199] - ChIP Grade (ab198977)
    Immunoprecipitation - Anti-CGBP antibody [EPR19199] - ChIP Grade (ab198977)

    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.

  • Anti-CGBP antibody [EPR19199] - ChIP Grade (ab198977)
    Anti-CGBP antibody [EPR19199] - ChIP Grade (ab198977)

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