Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-Nrf2 antibody [EP1808Y] - BSA and Azide free (ab180845)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP1808Y] to Nrf2 - BSA and Azide free
  • Suitable for: ICC, Flow Cyt, IP, IHC-P, WB, ChIP
  • Reacts with: Human

Overview

  • Product name

    Anti-Nrf2 antibody [EP1808Y] - BSA and Azide free
    See all Nrf2 primary antibodies

  • Description

    Rabbit monoclonal [EP1808Y] to Nrf2 - BSA and Azide free

  • Host species

    Rabbit

  • Specificity

    The expression of Nrf2 is stimulated by oxidative stress, electrophiles and chemical activators (PMID: 25761198, PMID: 27638861 and PMID: 28587109). Nrf2 antibody (ab62352) detects no signal in most untreated samples in WB. Stimuli treated samples are recommended.  We do not recommend using this product in western blot with tissue lysates, however some customers have used this antibody successfully using concentrated samples (see submitted abreviews).

  • Tested Applications & Species

    Application Species
    ChIP
    Human
    Flow Cyt
    Human
    IHC-P
    Human
    IP
    Human
    See all applications and species data

  • Immunogen

    Synthetic peptide within Human Nrf2 (C terminal). The exact sequence is proprietary.
    (Peptide available as ab167152)

  • Positive control

    • WB: MG-132 treated HeLa whole cell lysate, Saos-2, THP-1, MG-132 treated HepG2 whole cell lysate, MG-132 treated HCT-116 whole cell lysate. IHC-P: Human pancreatic carcinoma and kidney cancer tissues. ICC/IF: HepG2 and HeLa cells. Flow Cyt: HeLa cells. IP: SAOS-2 whole cell lysate. ChIP: Chromatin prepared from HepG2 cells.

  • General notes

    ab180845 is the carrier-free version of ab62352. 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

    ab180845 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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nrf2 antibody [EP1808Y] - BSA and Azide free (ab180845)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nrf2 antibody [EP1808Y] - BSA and Azide free (ab180845)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human pancreatic carcinoma tissue labelling Nrf2 with purified ab62352 at a dilution of 1/250. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9 (ab93684). Goat Anti-Rabbit IgG H&L (HRP) (ab97051) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab62352).

  • Immunocytochemistry - Anti-Nrf2 antibody [EP1808Y] - BSA and Azide free (ab180845)
    Immunocytochemistry - Anti-Nrf2 antibody [EP1808Y] - BSA and Azide free (ab180845)

    Clone EP1808Y (ab180845) has been successfully conjugated by Abcam. This image was generated using Anti-Nrf2 antibody [EP1808Y] (Alexa Fluor® 647). Please refer to ab194985 for protocol details.

    ab194985 staining Nrf2 in HeLa cells. The cells were fixed with 100% methanol (5 min), permeabilized in 0.1% Triton X-100 for 5 minutes and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated with ab194985 at a working dilution 1/100 (shown in red) and ab195887, Mouse monoclonal [DM1A] to alpha Tubulin (Alexa Fluor® 488, shown in green) at 2µg/ml overnight at +4°C. Nuclear DNA was labelled in blue with DAPI.

    This product gave a positive signal in 4% formaldehyde (10 min) fixed HeLa cells under the same testing conditions.

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

  • Flow Cytometry - Anti-Nrf2 antibody [EP1808Y] - BSA and Azide free (ab180845)
    Flow Cytometry - Anti-Nrf2 antibody [EP1808Y] - BSA and Azide free (ab180845)

    Clone EP1808Y (ab180845) has been successfully conjugated by Abcam. This image was generated using Anti-Nrf2 antibody [EP1808Y] (PE). Please refer to ab223926 for protocol details.

    Overlay histogram showing HeLa (human epithelial cell line from cervix adenocarcinoma) cells stained with ab223926 (red line). The cells were fixed with 4% formaldehyde (10 minutes) and then permeabilized with 0.1% PBS-Triton X-100 for 15 minutes. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab223926, 1/5000 dilution) for 30 minutes at 22°C.

    Isotype control antibody (black line) was Rabbit IgG (monoclonal) Phycoerythrin (ab209478) used at the same concentration and conditions as the primary antibody. Unlabeled sample (blue line) was also used as a control.

    Acquisition of >5,000 events were collected using a 50 mW Yellow/Green laser (561nm) and 586/15 bandpass filter.

    This antibody gave a positive signal in HeLa cells fixed with 80% methanol (5 minutes)/permeabilized with 0.1% PBS-Triton X-100 for 15 minutes used under the same conditions.

  • ChIP - Anti-Nrf2 antibody [EP1808Y] - BSA and Azide free (ab180845)
    ChIP - Anti-Nrf2 antibody [EP1808Y] - BSA and Azide free (ab180845)

    Chromatin was prepared from HepG2 cells according to the Abcam X-ChIP protocol. Cells were fixed with 1% formaldehyde for 10 minutes. The ChIP was performed with 25µg of chromatin, 2µg of ab62352 (blue), and 20µl of A/G sepharose bead slurry (10µl of sepharose A beads + 10µl of sepharose G beads). 2μg of rabbit normal IgG was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab62352).

  • Immunocytochemistry - Anti-Nrf2 antibody [EP1808Y] - BSA and Azide free (ab180845)
    Immunocytochemistry - Anti-Nrf2 antibody [EP1808Y] - BSA and Azide free (ab180845)

    Clone EP1808Y (ab180845) has been successfully conjugated by Abcam. This image was generated using Anti-Nrf2 antibody [EP1808Y] (Alexa Fluor® 488). Please refer to ab194984 for protocol details.

    ab194984 staining Nrf2 in HeLa cells. The cells were fixed with 4% formaldehyde (10 min), permeabilized in 0.1% Triton X-100 for 5 minutes and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated with ab194984 at a working dilution of 1/100 (shown in green) and ab195889, Mouse monoclonal [DM1A] to alpha Tubulin (Alexa Fluor® 594, shown in red) at 2µg/ml overnight at +4°C. Nuclear DNA was labelled in blue with DAPI.

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

  • Immunocytochemistry - Anti-Nrf2 antibody [EP1808Y] - BSA and Azide free (ab180845)
    Immunocytochemistry - Anti-Nrf2 antibody [EP1808Y] - BSA and Azide free (ab180845)

    Immunocytochemistry/Immunofluorescence analysis of HepG2 cells labelling Nrf2 with purified ab62352 at 1/500. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. The cells were co-stained with ab195889, an Alexa Fluor® 594-conjugated mouse anti-alpha tubulin antibody (1/200). Nuclei counterstained with DAPI (blue).

    Secondary antibody only control: PBS was used instead of the primary antibody as the negative control.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab62352).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nrf2 antibody [EP1808Y] - BSA and Azide free (ab180845)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nrf2 antibody [EP1808Y] - BSA and Azide free (ab180845) This image is courtesy of an Abreview submitted by Rudolf Jung.

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney cancer tissue sections labeling Nrf2 with ab62352 at 1/100 dilution. The tissue was fixed with paraformaldehyde and a heat mediated antigen retrival step was performed with TRIS-EDTA Buffer pH 9.0. Staining with ab62352 at 1/100 was carried out in a dilution buffer with blocking for 30 minutes at 20°C. A undiluted goat anti-rabbit HRP conjugated secondary antibody was used.  

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab62352).

  • Immunocytochemistry - Anti-Nrf2 antibody [EP1808Y] - BSA and Azide free (ab180845)
    Immunocytochemistry - Anti-Nrf2 antibody [EP1808Y] - BSA and Azide free (ab180845)

    Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling Nrf2 with purified ab62352 at 1/500. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody (1/1000) was used as the secondary antibody. Cells were counterstained with ab195889, anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594). DAPI was used to stain the nuclei blue.

    Secondary antibody only control: PBS was used instead of the primary antibody as the negative control.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab62352).

  • Immunoprecipitation - Anti-Nrf2 antibody [EP1808Y] - BSA and Azide free (ab180845)
    Immunoprecipitation - Anti-Nrf2 antibody [EP1808Y] - BSA and Azide free (ab180845)

    ab62352 (purified) at 1/30 immunoprecipitating Nrf2 in SAOS-2 whole cell lysate.

    Lane 1 (input): SAOS-2 whole cell lysate (10µg)
    Lane 2 (+): ab62352 + SAOS-2 whole cell lysate.
    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab62352 in SAOS-2 whole cell lysate.

    For western blotting, ab62352 was used at a dilution of 1/500 and VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

    Blocking and dilution buffer: 5% NFDM/TBST.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab62352).

  • Flow Cytometry - Anti-Nrf2 antibody [EP1808Y] - BSA and Azide free (ab180845)
    Flow Cytometry - Anti-Nrf2 antibody [EP1808Y] - BSA and Azide free (ab180845)

    Flow Cytometry analysis of HeLa cells labelling Nrf2 with purified ab62352 at a dilution of 1/60 (red). Cells were fixed with 4% paraformaldehyde. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody (1/2000) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG (ab172730). Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab62352).

  • Flow Cytometry - Anti-Nrf2 antibody [EP1808Y] - BSA and Azide free (ab180845)
    Flow Cytometry - Anti-Nrf2 antibody [EP1808Y] - BSA and Azide free (ab180845)

    Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labelling Nrf2 with ab62352 at 1/40 (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/2000) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab62352).

  • Anti-Nrf2 antibody [EP1808Y] - BSA and Azide free (ab180845)
    Anti-Nrf2 antibody [EP1808Y] - BSA and Azide free (ab180845)

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