Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-Vitamin D Receptor antibody [EPR4552] - ChIP Grade (ab109234)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR4552] to Vitamin D Receptor - ChIP Grade
  • Suitable for: WB, IP, ChIP
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-Vitamin D Receptor antibody [EPR4552] - ChIP Grade
    See all Vitamin D Receptor primary antibodies

  • Description

    Rabbit monoclonal [EPR4552] to Vitamin D Receptor - ChIP Grade

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    ChIP
    Human
    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: HeLa, U-937, T-47D and SKBR-3 cell lysates; Rat and mouse kidney tissue lysates. ChIP: Chromatin prepared from T-47D cells.

  • 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.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Properties

Images

  • Western blot - Anti-Vitamin D Receptor antibody [EPR4552] - ChIP Grade (ab109234)
    Western blot - Anti-Vitamin D Receptor antibody [EPR4552] - ChIP Grade (ab109234)
    All lanes : Anti-Vitamin D Receptor antibody [EPR4552] - ChIP Grade (ab109234) at 1/1000 dilution

    Lane 1 : Wild-type HeLa lysate
    Lane 2 : Vitamin D Receptor knockout HeLa lysate
    Lane 3 : U-937 lysate
    Lane 4 : SH-SY5Y lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 48 kDa



    Lanes 1-4: Merged signal (red and green). Green - ab109234 observed at 50 kDa. Red - loading control ab8245 observed at 37 kDa.

     ab109234 Anti-Vitamin D Receptor antibody [EPR4552] - ChIP Grade was shown to specifically react with Vitamin D Receptor in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265430 (knockout cell lysate ab257796) was used. Wild-type and Vitamin D Receptor knockout samples were subjected to SDS-PAGE. ab109234 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution 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.

     

     

  • ChIP - Anti-Vitamin D Receptor antibody [EPR4552] - ChIP Grade (ab109234)
    ChIP - Anti-Vitamin D Receptor antibody [EPR4552] - ChIP Grade (ab109234)

    Chromatin was prepared from T-47D cells according to the Abcam X-ChIP protocol. Cells were fixed with 1% formaldehyde for 10 minutes. The ChIP was performed with 25 µg of chromatin, 5 µg of ab109234 (red), and 20 µL of protein A/G sepharose beads slurry (10 µL of sepharose A beads + 10 µL of sepharose G beads). 5 μg of rabbit normal IgG was added to the beads as a control sample (grey). The immunoprecipitated DNA was quantified by real time PCR (SYBR Green chemistry) with primers to c-Fos.

  • Western blot - Anti-Vitamin D Receptor antibody [EPR4552] - ChIP Grade (ab109234)
    Western blot - Anti-Vitamin D Receptor antibody [EPR4552] - ChIP Grade (ab109234)
    Anti-Vitamin D Receptor antibody [EPR4552] - ChIP Grade (ab109234) at 1/5000 dilution (purified) + HeLa cell lysate at 20 µg

    Secondary
    Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 48 kDa
    Observed band size: 48 kDa



    Blocking and diluting buffer: 5% NFDM/TBST.

  • Immunoprecipitation - Anti-Vitamin D Receptor antibody [EPR4552] - ChIP Grade (ab109234)
    Immunoprecipitation - Anti-Vitamin D Receptor antibody [EPR4552] - ChIP Grade (ab109234)

    Lane 1 (input): T-47D (Human ductal breast epithelial tumor epithelial cell) whole cell lysate, 10 μg
    Lane 2(+): T-47D whole cell lysate
    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab109234 in T-47D whole cell lysate

    Ab109234 immunoprecipitating Vitamin D receptor in T-47D whole cell lysates. Capture antibody was used at a 1/30 dilution (2 μg in 0.35 mg lysates). For western blotting, primary antibody was used as ab109234 at 1/1000 dilution (0.62 μg/mL). VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.

  • Western blot - Anti-Vitamin D Receptor antibody [EPR4552] - ChIP Grade (ab109234)
    Western blot - Anti-Vitamin D Receptor antibody [EPR4552] - ChIP Grade (ab109234)
    Anti-Vitamin D Receptor antibody [EPR4552] - ChIP Grade (ab109234) at 1/5000 dilution (purified) + Mouse kidney tissue lysate at 20 µg

    Secondary
    Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 48 kDa
    Observed band size: 48 kDa



    Blocking and diluting buffer: 5% NFDM/TBST.

  • Western blot - Anti-Vitamin D Receptor antibody [EPR4552] - ChIP Grade (ab109234)
    Western blot - Anti-Vitamin D Receptor antibody [EPR4552] - ChIP Grade (ab109234)
    Anti-Vitamin D Receptor antibody [EPR4552] - ChIP Grade (ab109234) at 1/1000 dilution (purified) + Rat kidney tissue lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 48 kDa
    Observed band size: 48 kDa



    Blocking and diluting buffer: 5% NFDM/TBST.

  • Western blot - Anti-Vitamin D Receptor antibody [EPR4552] - ChIP Grade (ab109234)
    Western blot - Anti-Vitamin D Receptor antibody [EPR4552] - ChIP Grade (ab109234)
    All lanes : Anti-Vitamin D Receptor antibody [EPR4552] - ChIP Grade (ab109234) at 1/1000 dilution (unpurified)

    Lane 1 : T47D cell lysate
    Lane 2 : SKBR-3 cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : HRP-conjugated goat anti-rabbit IgG at 1/2000 dilution

    Predicted band size: 48 kDa
    Observed band size: 48 kDa

  • Anti-Vitamin D Receptor antibody [EPR4552] - ChIP Grade (ab109234)
    Anti-Vitamin D Receptor antibody [EPR4552] - ChIP Grade (ab109234)

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