Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)

Overview

  • Product name
    Rabbit IgG, polyclonal - Isotype Control (ChIP Grade)
    See all Rabbit isotype controls
  • Specificity
    ab171870 does not react with any antigen and was not produced using an immunogen.
  • Tested applications
    Suitable for: WB, ChIP, Flow Cytmore details

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer
    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituent: PBS
  • Concentration information loading...
  • Purity
    Protein G purified
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas
  • Alternative names
    • rabbit isotype control

Applications

Our Abpromise guarantee covers the use of ab171870 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
WB Use a concentration of 1 µg/ml.
ChIP Use a concentration of 1 µg/ml.
Flow Cyt Use at an assay dependent concentration.

Please note: This product should be diluted to the same concentration (not dilution) of the primary antibody to be used.

Images

  • Chromatin was prepared from HeLa cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25µg of chromatin, 2µg of ab171870 (blue), and 20µl of Protein A/G sepharose beads. No antibody was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Taqman approach for active and inactive loci, Sybr green approach for heterochromatic loci). Primers and probes are located in the first kb of the transcribed region.

  • Overlay histogram showing HeLa cells stained with ab75186 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Triton X-100 for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab75186, 0.05μg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150081) at 1/4000 dilution for 30 min at 22ºC.

    Isotype control antibody (black line) was rabbit IgG (polyclonal) (ab171870, 0.05μg/1x10cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control.

    Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

  • Lanes 1 - 6 : Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870) at 1 µg/ml
    Lanes 7 - 12 : Anti-beta Actin antibody (ab8227) at 1 µg/ml

    Lane 1 : Human liver tissue lysate - total protein (ab29889)
    Lane 2 : Liver (Mouse) Tissue Lysate
    Lane 3 : Liver (Rat) Tissue Lysate
    Lane 4 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 5 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
    Lane 6 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
    Lane 7 : Human liver tissue lysate - total protein (ab29889)
    Lane 8 : Liver (Mouse) Tissue Lysate
    Lane 9 : Liver (Rat) Tissue Lysate
    Lane 10 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 11 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
    Lane 12 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Exposure time : 2 minutes

    Please note that ab171870 in lanes 1-6 represents a negative control for Beta Actin, positively seen in lanes 7-12.

    This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 5% Bovine Serum Albumin before being incubated with ab171870 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.

  • All lanes : Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870) at 1 µg/ml

    Lane 1 : Liver (Human) Tissue Lysate - adult normal tissue
    Lane 2 : Brain (Mouse) Tissue Lysate
    Lane 3 : Rat Kidney Tissue Lysate
    Lane 4 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 5 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
    Lane 6 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Exposure time : 20 minutes

    This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 5% Bovine Serum Albumin before being incubated with ab171870 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.

References

This product has been referenced in:
  • Zhang D  et al. Non-CpG methylation by DNMT3B facilitates REST binding and gene silencing in developing mouse hearts. Nucleic Acids Res 45:3102-3115 (2017). ChIP . Read more (PubMed: 27956497) »
  • Roth-Walter F  et al. Janus-faced Acrolein prevents allergy but accelerates tumor growth by promoting immunoregulatory Foxp3+ cells: Mouse model for passive respiratory exposure. Sci Rep 7:45067 (2017). Read more (PubMed: 28332605) »

See all 22 Publications for this product

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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