Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-APE1 antibody [EPR18378-45] - BSA and Azide free (ab227062)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR18378-45] to APE1 - BSA and Azide free
  • Suitable for: WB, Flow Cyt, ICC/IF, ChIP, IHC-P
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-APE1 antibody [EPR18378-45] - BSA and Azide free
    See all APE1 primary antibodies

  • Description

    Rabbit monoclonal [EPR18378-45] to APE1 - BSA and Azide free

  • Host species

    Rabbit

  • Tested Applications & Species

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

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: WIld-type HAP1, HEK293, and HepG2 cell lysates. IHC-P: Human ovarian carcinoma tissue.

  • General notes

    Ab227062 is the carrier-free version of ab189474. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

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

    ab227062 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    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.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Properties

Images

  • Western blot - Anti-APE1 antibody [EPR18378-45] - BSA and Azide free (ab227062)
    Western blot - Anti-APE1 antibody [EPR18378-45] - BSA and Azide free (ab227062)
    All lanes : Anti-APE1 antibody [EPR18378-45] - ChIP Grade (ab189474) at 1/1000 dilution

    Lane 1 : Wild-type HAP1 whole cell lysate
    Lane 2 : APEX1 knockout HAP1 whole cell lysate
    Lane 3 : HepG2 whole cell lysate
    Lane 4 : HEK293 whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 35 kDa
    Observed band size: 37 kDa
    why is the actual band size different from the predicted?



    This data was developed using the same antibody clone in a different buffer formulation (ab189474).

    Lanes 1 - 4: Merged signal (red and green). Green - ab189474 observed at 37 kDa. Red - loading control, ab7291 (Mouse anti-Alpha Tubulin [DM1A] observed at 55kDa.

    ab189474 was shown to react with APEX1 in HAP1 wild-type cells in Western blot. Loss of signal was observed when APEX1 knockout sample was used. HAP1 wild-type and APEX1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% Milk in TBS-T (0.1% Tween®) before incubation with ab189474 and ab7291 (Mouse anti-Alpha Tubulin [DM1A] overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • ChIP - Anti-APE1 antibody [EPR18378-45] - BSA and Azide free (ab227062)
    ChIP - Anti-APE1 antibody [EPR18378-45] - BSA and Azide free (ab227062)

    Chromatin was prepared from HCT 116 cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25µg of chromatin, 5µg of ab189474 (blue), and 20µl of Protein A/G sepharose beads slurry (10µl of sepharose A beads + 10µl of sepharose G beads). 5μg of rabbit normal IgG was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (SYBR green approach).

    ChIP was performed according to the literature (PMID: 23874636).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab189474).

  • Flow Cytometry - Anti-APE1 antibody [EPR18378-45] - BSA and Azide free (ab227062)
    Flow Cytometry - Anti-APE1 antibody [EPR18378-45] - BSA and Azide free (ab227062)

    Flow cytometric analysis of 4% pasraformaldehyde-fixed, 90% methanol-permeabilized NIH/3T3 (mouse embryo fibroblast cell line) cell line labeling APE1 with ab189474 at 1/500 dilution (red) compared with a Isotype control details (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue).

    Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077), at 1/2000 dilution was used as the secondary antibody.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab189474).

  • Immunocytochemistry/ Immunofluorescence - Anti-APE1 antibody [EPR18378-45] - BSA and Azide free (ab227062)
    Immunocytochemistry/ Immunofluorescence - Anti-APE1 antibody [EPR18378-45] - BSA and Azide free (ab227062)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (mouse embryo fibroblast cell line) cells labeling APE1 with ab189474 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining on NIH/3T3 cell line.

    The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution.


    -ve control : PBS, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab189474).

  • Immunocytochemistry/ Immunofluorescence - Anti-APE1 antibody [EPR18378-45] - BSA and Azide free (ab227062)
    Immunocytochemistry/ Immunofluorescence - Anti-APE1 antibody [EPR18378-45] - BSA and Azide free (ab227062)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HCT 116 (human colorectal carcinoma cell line) cells labeling APE1 with ab189474 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining on HCT 116 cell line.

    The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution.


    -ve control : PBS, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab189474).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-APE1 antibody [EPR18378-45] - BSA and Azide free (ab227062)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-APE1 antibody [EPR18378-45] - BSA and Azide free (ab227062)

    Immunohistochemical analysis of paraffin-embedded rat liver tissue labeling APE1 with ab189474 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Mainly nuclear staining on hepatocytes of rat liver (PMID: 10643898) 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) ready to use.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab189474).

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-APE1 antibody [EPR18378-45] - BSA and Azide free (ab227062)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-APE1 antibody [EPR18378-45] - BSA and Azide free (ab227062)

    Immunohistochemical analysis of paraffin-embedded mouse liver tissue labeling APE1 with ab189474 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use.

    Immunohistochemical analysis of paraffin-embedded human ovarian cancer tissue labeling APE1 with ab189474 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Nuclear staining on tumor cells of human ovarian carcinoma (PMID: 20087352) 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) ready to use.

    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) ready to use.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab189474).

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-APE1 antibody [EPR18378-45] - BSA and Azide free (ab227062)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-APE1 antibody [EPR18378-45] - BSA and Azide free (ab227062)

    Immunohistochemical analysis of paraffin-embedded human ovarian cancer tissue labeling APE1 with ab189474 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Nuclear staining on tumor cells of human ovarian carcinoma (PMID: 20087352) 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) ready to use.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab189474).

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Anti-APE1 antibody [EPR18378-45] - BSA and Azide free (ab227062)
    Anti-APE1 antibody [EPR18378-45] - BSA and Azide free (ab227062)

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