RabMAb

Anti-Heme Oxygenase 1 antibody [EPR18161-128] (ab189491)

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Overview

  • Product name
    Anti-Heme Oxygenase 1 antibody [EPR18161-128]
    See all Heme Oxygenase 1 primary antibodies
  • Description
    Rabbit monoclonal [EPR18161-128] to Heme Oxygenase 1
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IHC-P, Flow Cyt, IP, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment within Mouse Heme Oxygenase 1 aa 1 to the C-terminus. The exact sequence is proprietary.
    Database link: P14901

    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI
  • Positive control
    • WB: Human, mouse and rat spleen lysate. NIH/3T3 and HeLa cell lysate. IHC-P: Human, mouse and rat liver tissue. ICC/IF: HeLa and NIH/3T3 cells. Flow Cytometry: HeLa and NIH/3T3 cells. IP: NIH/3T3 whole cell lysate.
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab189491 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
WB 1/2000. Predicted molecular weight: 33 kDa.
IHC-P 1/20000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Flow Cyt 1/50.
IP 1/30.
ICC/IF 1/250.

Target

  • Function
    Heme oxygenase cleaves the heme ring at the alpha methene bridge to form biliverdin. Biliverdin is subsequently converted to bilirubin by biliverdin reductase. Under physiological conditions, the activity of heme oxygenase is highest in the spleen, where senescent erythrocytes are sequestrated and destroyed.
  • Sequence similarities
    Belongs to the heme oxygenase family.
  • Cellular localization
    Microsome. Endoplasmic reticulum.
  • Information by UniProt
  • Database links
    see all
  • Alternative names
    • 32 kD antibody
    • bK286B10 antibody
    • D8Wsu38e antibody
    • heat shock protein 32 kD antibody
    • heat shock protein 32kD antibody
    • Heat shock protein antibody
    • Heme oxygenase (decycling) 1 antibody
    • Heme oxygenase 1 antibody
    • Hemox antibody
    • HMOX 1 antibody
    • Hmox antibody
    • Hmox1 antibody
    • HMOX1_HUMAN antibody
    • HO 1 antibody
    • HO antibody
    • HO-1 antibody
    • HO1 antibody
    • Hsp32 antibody
    see all

Images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Heme Oxygenase 1 antibody [EPR18161-128] (ab189491)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Heme Oxygenase 1 antibody [EPR18161-128] (ab189491)

    Immunohistochemical analysis of paraffin embedded human liver tissue labeling Heme Oxygenase 1 with ab189491 at 1/20,000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining on Kupffer cells of human liver (PMID: 9449694) is observed. Counter stained with hematoxylin.
    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

    Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

  • Immunocytochemistry/ Immunofluorescence - Anti-Heme Oxygenase 1 antibody [EPR18161-128] (ab189491)
    Immunocytochemistry/ Immunofluorescence - Anti-Heme Oxygenase 1 antibody [EPR18161-128] (ab189491)

    Immunofluorescent analysis of 4% paraformaldehyde fixed, 0.1% Triton X-100 permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labeling Heme Oxygenase 1 with ab189491 at 1/250 dilution, followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on HeLa cells. Details of counterstains: ab195889  Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at a 1/200 dilution; DAPI for nuclei.
    The negative controls are as follows: Secondary antibody only for control.

  • Immunocytochemistry/ Immunofluorescence - Anti-Heme Oxygenase 1 antibody [EPR18161-128] (ab189491)
    Immunocytochemistry/ Immunofluorescence - Anti-Heme Oxygenase 1 antibody [EPR18161-128] (ab189491)

    Immunofluorescent analysis of 4% paraformaldehyde fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labeling Heme Oxygenase 1 with ab189491 at 1/250 dilution, followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on NIH/3T3 cells. Details of counterstains: ab195889  Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at a 1/200 dilution; DAPI for nuclei.
    The negative controls are as follows: Secondary antibody only for control.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Heme Oxygenase 1 antibody [EPR18161-128] (ab189491)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Heme Oxygenase 1 antibody [EPR18161-128] (ab189491)

    Immunohistochemical analysis of paraffin embedded mouse liver tissue labeling Heme Oxygenase 1 with ab189491 at 1/20000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining on Kupffer cells of mouse liver (PMID: 9449694) is observed. Counter stained with hematoxylin.
    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

    Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Heme Oxygenase 1 antibody [EPR18161-128] (ab189491)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Heme Oxygenase 1 antibody [EPR18161-128] (ab189491)

    Immunohistochemical analysis of paraffin embedded rat liver tissue labeling Heme Oxygenase 1 with ab189491 at 1/20,000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining on Kupffer cells of rat liver (PMID: 9449694) is observed. Counter stained with hematoxylin.
    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

    Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

  • Western blot - Anti-Heme Oxygenase 1 antibody [EPR18161-128] (ab189491)
    Western blot - Anti-Heme Oxygenase 1 antibody [EPR18161-128] (ab189491)
    Anti-Heme Oxygenase 1 antibody [EPR18161-128] (ab189491) at 1/2000 dilution + Human spleen lysate at 10 µg

    Secondary
    VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/4000 dilution

    Predicted band size: 33 kDa
    Observed band size: 28, 32 kDa (why is the actual band size different from the predicted?)


    Exposure time: 3 minutes


    Blocking: 5% NFDM/TBST.

    The molecular weight observed is consistent with what has been described in the literature (PMID: 18400743).

    The lower band observed is a truncated form of Heme Oxygenase 1 (PMID: 17430897).

  • Western blot - Anti-Heme Oxygenase 1 antibody [EPR18161-128] (ab189491)
    Western blot - Anti-Heme Oxygenase 1 antibody [EPR18161-128] (ab189491)
    All lanes : Anti-Heme Oxygenase 1 antibody [EPR18161-128] (ab189491) at 1/5000 dilution

    Lane 1 : NIH/3T3 (mouse embryonic fibroblast) lysate
    Lane 2 : Rat spleen lysate
    Lane 3 : HeLa (human cervix adenocarcinoma epithelial cell) lysate
    Lane 4 : Mouse spleen lysate

    Lysates/proteins at 20 µg per lane.

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

    Predicted band size: 33 kDa
    Observed band size: 28, 32 kDa (why is the actual band size different from the predicted?)



    Blocking: 5% NFDM/TBST.

    Exposure time:

    Lanes 1,2 and 3: 2 seconds;

    Lane 4: 1 second

    The molecular weight observed is consistent with what has been described in the literature (PMID: 18400743).

    The lower band observed is a truncated form of Heme Oxygenase 1 (PMID: 17430897).

  • Flow Cytometry - Anti-Heme Oxygenase 1 antibody [EPR18161-128] (ab189491)
    Flow Cytometry - Anti-Heme Oxygenase 1 antibody [EPR18161-128] (ab189491)

    Flow cytometric analysis of 90% methanol/ 4% paraformaldehyde fixed NIH/3T3 (mouse embryonic fibroblast) cell line labeling Heme Oxygenase 1  with ab189491 at 1/50 (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077), at 1/2000 dilution was used as the secondary antibody.

  • Flow Cytometry - Anti-Heme Oxygenase 1 antibody [EPR18161-128] (ab189491)
    Flow Cytometry - Anti-Heme Oxygenase 1 antibody [EPR18161-128] (ab189491)

    Flow cytometric analysis of 90% methanol/4% paraformaldehyde fixed HeLa (human cervix adenocarcinoma epithelial cell) cell line labeling Heme Oxygenase 1  with ab189491 at 1/50 (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077), at 1/2000 dilution was used as the secondary antibody.

  • Immunoprecipitation - Anti-Heme Oxygenase 1 antibody [EPR18161-128] (ab189491)
    Immunoprecipitation - Anti-Heme Oxygenase 1 antibody [EPR18161-128] (ab189491)

    Heme Oxygenase 1 was immunoprecipitated from 0.35 mg of NIH/3T3 (mouse embryonic fibroblast) whole cell lysate with ab189491 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab189491 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366), was used as secondary antibody at 1/10,000 dilution.
    Lane 1: NIH/3T3 (mouse embryonic fibroblast) whole cell lysate  10 μg (Input).
    Lane 2: NIH/3T3 whole cell lysate (+).
    Lane 3: Rabbit monoclonal IgG (ab172730) instead of  ab189491 in NIH/3T3 whole cell lysate (-).
    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    The truncated form of Heme Oxygenase 1 is described in the literature (PMID: 17430897).

References

ab189491 has not yet been referenced specifically in any publications.

Publishing research using ab189491? Please let us know so that we can cite the reference in this datasheet.

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