Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-EED antibody [EPR23043-5] - BSA and Azide free (ab264566)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR23043-5] to EED - BSA and Azide free
  • Suitable for: IP, ChIP-sequencing, WB, ChIP
  • Knockout validated
  • Reacts with: Mouse, Human

Overview

  • Product name

    Anti-EED antibody [EPR23043-5] - BSA and Azide free
    See all EED primary antibodies

  • Description

    Rabbit monoclonal [EPR23043-5] to EED - BSA and Azide free

  • Host species

    Rabbit

  • Specificity

    ab240650 detects an unknown band close to the target bands in cytoplasm.

  • Tested applications

    Suitable for: IP, ChIP-sequencing, WB, ChIPmore details
    Unsuitable for: ICC/IF or IHC-P

  • Species reactivity

    Reacts with: Mouse, Human

  • Immunogen

    This product was produced with the following immunogens:
    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

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

  • Positive control

    • WB: Wild type HAP1, K562, 293T, NIH/3T3 and C2C12 lysates. IP: K562 cells.

  • General notes

    ab264566 is the carrier-free version of ab240650.

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. 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

Images

  • Western blot - Anti-EED antibody [EPR23043-5] - ChIP Grade – BSA and Azide free (ab264566)
    Western blot - Anti-EED antibody [EPR23043-5] - ChIP Grade – BSA and Azide free (ab264566)
    All lanes : Anti-EED antibody [EPR23043-5] - ChIP Grade (ab240650) at 1/1000 dilution

    Lanes 1-2 : 293T (human embryonic kidney epithelial cell) whole cell lysate
    Lane 3 : K562 (human chronic myelogenous leukemia lymphoblast), whole cell lysate
    Lane 4 : 293T (human embryonic kidney epithelial cell), whole cell lysate
    Lane 5 : NIH/3T3 (mouse embryonic fibroblast), whole cell lysate
    Lane 6 : C2C12 (mouse myoblasts myoblast), whole cell lysate

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/20000 dilution

    Predicted band size: 50 kDa



    This data was developed using ab240650, the same antibody clone in a different buffer formulation. 

    Blocking and diluting buffer and concentration: ab240650 was shown to specifically react with EED in wild-type HAP1 cells as signal was lost in EED knockout cells. Wild-type and EED knockout samples were subjected to SDS-PAGE. ab240650 and ab181602 (Rabbit anti-GAPDH loading control) were incubated 1 hour at room temperature at 1/1000 dilution and 1/200, 000 dilution respectively. Blots were developed with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) secondary antibody at 1/100, 000 dilution for 1 hour at room temperature before imaging.

     

    The molecular weight observed is consistent with what has been described in the literature (PMID:27578866, 9584199). EED cDNA encodes 441-aa-long protein and 535-aa-long protein.

     

    Lanes 1-3: 15 seconds; Lanes 4-6: 37 seconds.

    Exposure time: 

  • Immunoprecipitation - Anti-EED antibody [EPR23043-5] - ChIP Grade – BSA and Azide free (ab264566)
    Immunoprecipitation - Anti-EED antibody [EPR23043-5] - ChIP Grade – BSA and Azide free (ab264566)

    This data was developed using ab240650, the same antibody clone in a different buffer formulation.

    EED was immunoprecipitated from 0.35 mg K562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate 10ug with ab240650 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab240650 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

    Lane 1: K562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate 10ug

    Lane 2: ab240650 IP in K562 whole cell lysate

    Lane 3:Rabbit monoclonal IgG (ab172730) instead of ab240650 in K562 whole cell lysate

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

    Exposure time: 1 minutes

  • Western blot - Anti-EED antibody [EPR23043-5] - ChIP Grade – BSA and Azide free (ab264566)
    Western blot - Anti-EED antibody [EPR23043-5] - ChIP Grade – BSA and Azide free (ab264566)
    All lanes : Anti-EED antibody [EPR23043-5] - ChIP Grade (ab240650) at 1/1000 dilution

    Lane 1 : Wild type HAP1 whole cell lysate
    Lane 2 : EED knockout HAP1 whole cell lysate
    Lane 3 : K562 (human chronic myelogenous leukemia lymphoblast), whole cell lysate
    Lane 4 : 293T (human embryonic kidney epithelial cell), whole cell lysate
    Lane 5 : NIH/3T3 (mouse embryonic fibroblast), whole cell lysate
    Lane 6 : C2C12 (mouse myoblasts myoblast), whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 50 kDa
    Observed band size: 50-70 kDa
    why is the actual band size different from the predicted?



    Blocking and diluting buffer and concentration: 5% NFDM/TBST ab240650 was shown to specifically react with EED in wild-type HAP1 cells as signal was lost in EED knockout cells. Wild-type and EED knockout samples were subjected to SDS-PAGE. ab240650 and ab181602 (Rabbit anti-GAPDH loading control) were incubated 1 hour at room temperature at 1/1000 dilution and 1/200,000 dilution respectively. Blots were developed with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) secondary antibody at 1/100,000 dilution for 1 hour at room temperature before imaging.  The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID:27578866, 9584199). EED cDNA encodes 441-aa-long protein and 535-aa-long protein. 

    Exposure time: Lanes 1-3: 15 seconds Lanes 4-6: 37 seconds

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, glycerol, azide and BSA (ab240650).

  • Immunoprecipitation - Anti-EED antibody [EPR23043-5] - BSA and Azide free (ab264566)
    Immunoprecipitation - Anti-EED antibody [EPR23043-5] - BSA and Azide free (ab264566)

    EED was immunoprecipitated from 0.35 mg K562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate with ab240650 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab240650 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.

    Lane 1: K562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate 10 µg

    Lane 2: ab240650 IP in K562 whole cell lysate

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab240650 in K562 whole cell lysate

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

    Exposure time: 1 min

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, glycerol, azide and BSA (ab240650).

  • ChIP - Anti-EED antibody [EPR23043-5] - BSA and Azide free (ab264566)
    ChIP - Anti-EED antibody [EPR23043-5] - BSA and Azide free (ab264566)

    Chromatin was prepared from NT2/D1 cells according to the Abcam Dual-X-ChIP protocol*. Cells were fixed with 1.5 mM EGS for 30mins and then formaldehyde for 10min.

    The ChIP was performed with 25 µg of chromatin, 5 µg of ab240650 (red), or 5 µg of rabbit normal IgG ab172730 (gray) and 20 µl of Protein A/G sepharose beads.  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 commercial primers from Millipore (Cat. No.: 17-10034) and CST (85322S)

    *https://www.abcam.com/resources?keywords=X%20ChIP%20protocol

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, glycerol, azide and BSA (ab240650).

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