RecombinantRabMAb

Recombinant Anti-EHMT2/G9A + EHMT1/GLP antibody [EPR18667] (ab194299)

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Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR18667] to EHMT2/G9A + EHMT1/GLP
  • Suitable for: Flow Cyt, IHC-P, WB, ICC/IF
  • Reacts with: Human

Overview

  • Product name

    Anti-EHMT2/G9A + EHMT1/GLP antibody [EPR18667]
    See all EHMT2/G9A+EHMT1/GLP primary antibodies

  • Description

    Rabbit monoclonal [EPR18667] to EHMT2/G9A + EHMT1/GLP

  • Host species

    Rabbit

  • Tested applications

    Suitable for: Flow Cyt, IHC-P, WB, ICC/IFmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Recombinant fragment within Human EHMT1/GLP aa 1050-1250. The exact sequence is proprietary.
    Database link: Q9H9B1

  • Positive control

    • WB: Human fetal kidney lysate; HEK-293, HeLa, K562, MCF7 and PC-3 whole cell lysates; EHMT2 recombinant protein; IHC-P: Human tonsil tissue; ICC/IF: HeLa and NIH/3T3 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.

    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

Applications

Our Abpromise guarantee covers the use of ab194299 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
Flow Cyt Use at an assay dependent concentration.
IHC-P 1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB 1/1000. Detects a band of approximately 170 kDa (predicted molecular weight: 141, 132 kDa).
ICC/IF 1/1000.

Target

Images

  • Western blot - Anti-EHMT2/G9A + EHMT1/GLP antibody [EPR18667] (ab194299)
    Western blot - Anti-EHMT2/G9A + EHMT1/GLP antibody [EPR18667] (ab194299)
    Anti-EHMT2/G9A + EHMT1/GLP antibody [EPR18667] (ab194299) at 1/1000 dilution + Human fetal kidney lysate at 10 µg

    Secondary
    Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/10000 dilution

    Developed using the ECL technique.

    Predicted band size: 141, 132 kDa
    Observed band size: 170 kDa
    why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Flow Cytometry - Anti-EHMT2/G9A + EHMT1/GLP antibody [EPR18667] (ab194299)
    Flow Cytometry - Anti-EHMT2/G9A + EHMT1/GLP antibody [EPR18667] (ab194299)

    Flow cytometry analysis of HeLa (human cervix adenocarcinoma) cells labelling EHMT2/G9A + EHMT1/GLP (red) with purified ab194299 at a dilution of 1/150. Goat anti rabbit IgG (Alexa Fluor® 488) was used as the secondary antibody at 1/2000. Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Isotype control antibody was Rabbit monoclonal IgG (black). The blue line shows cells without incubation with primary antibody and secondary antibody.

  • Western blot - Anti-EHMT2/G9A + EHMT1/GLP antibody [EPR18667] (ab194299)
    Western blot - Anti-EHMT2/G9A + EHMT1/GLP antibody [EPR18667] (ab194299)
    All lanes : Anti-EHMT2/G9A + EHMT1/GLP antibody [EPR18667] (ab194299) at 1/5000 dilution

    Lane 1 : HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
    Lane 2 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 3 : K562 (Human chronic myelogenous leukemia cell line from bone marrow) 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: 141, 132 kDa
    Observed band size: 170 kDa why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lanes 1 & 2: 20 seconds; Lane 3: 30 seconds.

  • Western blot - Anti-EHMT2/G9A + EHMT1/GLP antibody [EPR18667] (ab194299)
    Western blot - Anti-EHMT2/G9A + EHMT1/GLP antibody [EPR18667] (ab194299)
    Lane 1 : Anti-EHMT2/G9A + EHMT1/GLP antibody [EPR18667] (ab194299) at 1/5000 dilution
    Lane 2 : Anti His-Tag®.

    All lanes : EHMT2 recombinant protein

    Lysates/proteins at 0.01 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Developed using the ECL technique.

    Predicted band size: 141, 132 kDa
    Observed band size: 30,60 kDa why is the actual band size different from the predicted?


    Exposure time: 5 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    EHMT2 recombinant protein fragment with His-Tag®.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EHMT2/G9A + EHMT1/GLP antibody [EPR18667] (ab194299)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EHMT2/G9A + EHMT1/GLP antibody [EPR18667] (ab194299)

    Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling EHMT2/G9A + EHMT1/GLP with ab194299 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on human tonsil 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-EHMT2/G9A + EHMT1/GLP antibody [EPR18667] (ab194299)
    Immunocytochemistry/ Immunofluorescence - Anti-EHMT2/G9A + EHMT1/GLP antibody [EPR18667] (ab194299)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling EHMT2/G9A + EHMT1/GLP with ab194299 at 1/1000 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 counter stain is DAPI (blue).

    Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/1000 dilution (red).

    The negative controls are as follows:
    -ve control 1: ab194299 at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) at 1/1000 dilution.
    -ve control 2: Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.

  • Immunocytochemistry/ Immunofluorescence - Anti-EHMT2/G9A + EHMT1/GLP antibody [EPR18667] (ab194299)
    Immunocytochemistry/ Immunofluorescence - Anti-EHMT2/G9A + EHMT1/GLP antibody [EPR18667] (ab194299)

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% tritonX-100 permeabilized NIH/3T3 (Mouse embryonic fibroblast cell line) cells labeling EHMT1/GLP + EHMT2/G9A with ab194299 at 1/1000 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. The nuclear counter stain is DAPI (blue).

    Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/1000 dilution (red).
    The negative controls are as follows:
    -ve control 1: ab194299 at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary at 1/1000 dilution.
    -ve control 2: Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary at 1/1000 dilution.

     

  • Anti-EHMT2/G9A + EHMT1/GLP antibody [EPR18667] (ab194299)
    Anti-EHMT2/G9A + EHMT1/GLP antibody [EPR18667] (ab194299)

References (0)

Publishing research using ab194299? Please let us know so that we can cite the reference in this datasheet.

ab194299 has not yet been referenced specifically in any publications.

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