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    heme-oxygenase-1-antibody-epr1390y-ab68477.pdf

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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Heme Oxygenase 1 antibody [EPR1390Y] (ab68477)

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  • SDS
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Western blot - Anti-Heme Oxygenase 1 antibody [EPR1390Y] (ab68477)
  • Immunocytochemistry/ Immunofluorescence - Anti-Heme Oxygenase 1 antibody [EPR1390Y] (ab68477)
  • Western blot - Anti-Heme Oxygenase 1 antibody [EPR1390Y] (ab68477)
  • Western blot - Anti-Heme Oxygenase 1 antibody [EPR1390Y] (ab68477)
  • Flow Cytometry (Intracellular) - Anti-Heme Oxygenase 1 antibody [EPR1390Y] (ab68477)
  • Immunoprecipitation - Anti-Heme Oxygenase 1 antibody [EPR1390Y] (ab68477)
  • Western blot - Anti-Heme Oxygenase 1 antibody [EPR1390Y] (ab68477)
  • Anti-Heme Oxygenase 1 antibody [EPR1390Y] (ab68477)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR1390Y] to Heme Oxygenase 1
  • Suitable for: Flow Cyt (Intra), ICC/IF, WB, IP
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 594 Carrier Free

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Secondary
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Knockout
Product image
Human HMOX1 (Heme Oxygenase 1) knockout A549 cell line (ab269503)

View more associated products

Overview

  • Product name

    Anti-Heme Oxygenase 1 antibody [EPR1390Y]
    See all Heme Oxygenase 1 primary antibodies
  • Description

    Rabbit monoclonal [EPR1390Y] to Heme Oxygenase 1
  • Host species

    Rabbit
  • Tested applications

    Suitable for: Flow Cyt (Intra), ICC/IF, WB, IPmore details
    Unsuitable for: IHC-P
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

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

  • Positive control

    • HepG2, A549, rat kidney, rat spleen, mouse kidney cell lysate.
  • 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.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: 40% Glycerol (glycerin, glycerine), 0.05% BSA, 59% PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR1390Y
  • Isotype

    IgG
  • Research areas

    • Neuroscience
    • Neurology process
    • Neurodegenerative disease
    • Alzheimer's disease
    • Other
    • Cardiovascular
    • Blood
    • Platelets
    • Signal Transduction
    • Signaling Pathway
    • Nuclear Signaling
    • NFkB Pathway
    • Epigenetics and Nuclear Signaling
    • Nuclear Signaling Pathways
    • NFkB pathway
    • Cardiovascular
    • Atherosclerosis
    • Vascular Inflammation
    • Inflammatory mediators
    • Cardiovascular
    • Vasculature
    • Endothelium
    • Cancer
    • Cancer Metabolism
    • Response to hypoxia
    • Metabolism
    • Pathways and Processes
    • Metabolism processes
    • Hypoxia
    • Metabolism
    • Types of disease
    • Neurodegenerative disease
    • Metabolism
    • Types of disease
    • Heart disease

Associated products

  • Alternative Versions

    • Alexa Fluor® 488 Anti-Heme Oxygenase 1 antibody [EPR1390Y] (ab214643)
    • Alexa Fluor® 594 Anti-Heme Oxygenase 1 antibody [EPR1390Y] (ab215098)
    • Anti-Heme Oxygenase 1 antibody [EPR1390Y] - BSA and Azide free (ab221215)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human HMOX1 (Heme Oxygenase 1) knockout A549 cell line (ab269503)
  • KO cell lysates

    • Human HMOX1 (Heme Oxygenase 1) knockout A549 cell lysate (ab269665)
  • Positive Controls

    • A549 whole cell lysate (ab7910)
  • Recombinant Protein

    • Recombinant Human Heme Oxygenase 1 protein (ab85243)
  • Related Products

    • Prestained Protein Ladder – Broad molecular weight (10-245 kDa) (ab116028)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab68477 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 (Intra)
Use at an assay dependent concentration.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ICC/IF (1)
1/100 - 1/250.
WB (4)
1/10000 - 1/50000. Detects a band of approximately 33 kDa (predicted molecular weight: 33 kDa).
IP
1/20.
Notes
Flow Cyt (Intra)
Use at an assay dependent concentration.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ICC/IF
1/100 - 1/250.
WB
1/10000 - 1/50000. Detects a band of approximately 33 kDa (predicted molecular weight: 33 kDa).
IP
1/20.
Application notes
Is unsuitable for IHC-P.

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.
  • Target information above from: UniProt accession P09601 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 3162 Human
    • Entrez Gene: 15368 Mouse
    • Entrez Gene: 24451 Rat
    • Omim: 141250 Human
    • SwissProt: P09601 Human
    • SwissProt: P14901 Mouse
    • SwissProt: P06762 Rat
    • Unigene: 517581 Human
    • Unigene: 276389 Mouse
    • Unigene: 3160 Rat
    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

  • Western blot - Anti-Heme Oxygenase 1 antibody [EPR1390Y] (ab68477)
    Western blot - Anti-Heme Oxygenase 1 antibody [EPR1390Y] (ab68477)
    All lanes : Anti-Heme Oxygenase 1 antibody [EPR1390Y] (ab68477) at 1/10000 dilution

    Lane 1 : Wild-type A549 cell lysate
    Lane 2 : HMOX1 knockout A549 cell lysate
    Lane 3 : Human Spleen tissue lysate
    Lane 4 : HL-60 cell lysate
    Lane 5 : MCF7 cell lysate
    Lane 6 : HeLa cell lysate
    Lane 7 : A549 cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 33 kDa
    Observed band size: 33 kDa



    Lanes 1 - 7: Merged signal (red and green). Green - ab68477 observed at 33 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.

    ab68477 was shown to react with Heme Oxygenase 1 in wild-type A549 cells in Western blot with loss of signal observed in HMOX1 knockout cell line ab269503 (knockout cell lysate ab269665). Wild-type A549 and HMOX1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab68477 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 °C at a 1 in 10000 dilution and a 1 in 20000 dilution respectively. Blots were incubated 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 h at room temperature before imaging.

  • Immunocytochemistry/ Immunofluorescence - Anti-Heme Oxygenase 1 antibody [EPR1390Y] (ab68477)
    Immunocytochemistry/ Immunofluorescence - Anti-Heme Oxygenase 1 antibody [EPR1390Y] (ab68477)

    Immunocytochemistry/Immunofluorescence analysis of HepG2 (Human liver hepatocellular carcinoma cell line) cells labeling Heme Oxygenase 1 with purified ab68477 at 1/100. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. A goat anti rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody at a dilution of 1/1000. DAPI was used as a nuclear counterstain.

    Negative control 1: PBS only.

  • Western blot - Anti-Heme Oxygenase 1 antibody [EPR1390Y] (ab68477)
    Western blot - Anti-Heme Oxygenase 1 antibody [EPR1390Y] (ab68477)
    All lanes : Anti-Heme Oxygenase 1 antibody [EPR1390Y] (ab68477) at 1/1000 dilution

    Lane 1 : Hek293
    Lane 2 : HL60
    Lane 3 : HeLa
    Lane 4 : A549
    Lane 5 : Hu spleen
    Lane 6 : Ms spleen
    Lane 7 : Rt spleen

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : IRDye® 800CW Goat anti Rabbit

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



    Hek293 & HL60 presumed negative or very low expression.

    Loading control GAPDH at 38kDa

  • Western blot - Anti-Heme Oxygenase 1 antibody [EPR1390Y] (ab68477)
    Western blot - Anti-Heme Oxygenase 1 antibody [EPR1390Y] (ab68477)
    All lanes : Anti-Heme Oxygenase 1 antibody [EPR1390Y] (ab68477) at 1/1000 dilution

    Lane 1 : HEK-293 (Human embryonic kidney epithelial cell) whole cell lysates with 5% NFDM/TBST
    Lane 2 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates with 5% NFDM/TBST
    Lane 3 : A549 (Human lung carcinoma epithelial cell) whole cell lysates with 5% NFDM/TBST
    Lane 4 : Mouse spleen lysates with 5% NFDM/TBST

    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: 33 kDa
    Observed band size: 33 kDa


    Exposure time: 10 seconds


    We are unsure how to define the extra bands.

  • Flow Cytometry (Intracellular) - Anti-Heme Oxygenase 1 antibody [EPR1390Y] (ab68477)
    Flow Cytometry (Intracellular) - Anti-Heme Oxygenase 1 antibody [EPR1390Y] (ab68477)

    Intracellular Flow Cytometry analysis of A549 (human lung carcinoma) cells labeling with purified ab68477 at 1/200 dilution (1ug/ml) (Red). Cells were fixed with4% paraformaldehydeand permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488) (ab150077) (1/2000 dilution) was used as the secondary antibody.Rabbit monoclonal IgG (Black) (ab172730) was used as a isotype control.Cell without incubation with primary antibody and secondary antibody (Blue) were used as unlabeled control. 

     

     

  • Immunoprecipitation - Anti-Heme Oxygenase 1 antibody [EPR1390Y] (ab68477)
    Immunoprecipitation - Anti-Heme Oxygenase 1 antibody [EPR1390Y] (ab68477)

    ab68477 (purified) at 1/20 immunoprecipitating Heme Oxygenase 1 in A549 (Human lung carcinoma cell line) whole cell lysate.

    Lane 1 (input): A549 whole cell lysate (10ug).

    Lane 2 (+): ab68477 + A549 whole cell lysate.

    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab133267 in HeLa whole cell lysate.

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • Western blot - Anti-Heme Oxygenase 1 antibody [EPR1390Y] (ab68477)
    Western blot - Anti-Heme Oxygenase 1 antibody [EPR1390Y] (ab68477)
    All lanes : Anti-Heme Oxygenase 1 antibody [EPR1390Y] (ab68477) at 1/20000 dilution (purified)

    Lane 1 : Rat kidney lysate
    Lane 2 : Rat spleen lysate
    Lane 3 : Mouse kidney lysate

    Lysates/proteins at 20 µg per lane.

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

    Predicted band size: 33 kDa
    Observed band size: 33 kDa



    Blocking and dilution buffer: 5% NFDM/TBST

  • Anti-Heme Oxygenase 1 antibody [EPR1390Y] (ab68477)
    Anti-Heme Oxygenase 1 antibody [EPR1390Y] (ab68477)

Protocols

  • Flow cytometry protocols
  • Immunoprecipitation protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (110)

Publishing research using ab68477? Please let us know so that we can cite the reference in this datasheet.

ab68477 has been referenced in 110 publications.

  • Zhang WB  et al. Protective Effects of Oroxylin A against Doxorubicin-Induced Cardiotoxicity via the Activation of Sirt1 in Mice. Oxid Med Cell Longev 2021:6610543 (2021). PubMed: 33542782
  • Bakalarz D  et al. Organic carbon monoxide prodrug, BW-CO-111, in protection against chemically-induced gastric mucosal damage. Acta Pharm Sin B 11:456-475 (2021). PubMed: 33643824
  • Weng Q  et al. Catalytic activity tunable ceria nanoparticles prevent chemotherapy-induced acute kidney injury without interference with chemotherapeutics. Nat Commun 12:1436 (2021). PubMed: 33664241
  • Deng S  et al. Albumin Reduces Oxidative Stress and Neuronal Apoptosis via the ERK/Nrf2/HO-1 Pathway after Intracerebral Hemorrhage in Rats. Oxid Med Cell Longev 2021:8891373 (2021). PubMed: 33708336
  • Qi B  et al. Brazilin prevents against myocardial ischemia-reperfusion injury through the modulation of Nrf2 via the PKC signaling pathway. Ann Transl Med 9:312 (2021). PubMed: 33708939
View all Publications for this product

Customer reviews and Q&As

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1-5 of 5 Abreviews

Western blot abreview for Anti-Heme Oxygenase 1 antibody [EPR1390Y]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (Skov-3 cells)
Gel Running Conditions
Non-reduced Denaturing (10)
Loading amount
60 µg
Treatment
50 µM hemin
Specification
Skov-3 cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Dec 08 2021

Immunocytochemistry/ Immunofluorescence abreview for Anti-Heme Oxygenase 1 antibody [EPR1390Y]

Excellent
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (MDA-MB-231)
Permeabilization
Yes - 0.2% Triton X-100
Specification
MDA-MB-231
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C
Fixative
Paraformaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

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Verified customer

Submitted Jan 13 2021

Western blot abreview for Anti-Heme Oxygenase 1 antibody [EPR1390Y]

Good
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Tissue lysate - whole (brain)
Gel Running Conditions
Reduced Denaturing (12)
Loading amount
50 µg
Specification
brain
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted May 13 2016

Western blot abreview for Anti-Heme Oxygenase 1 antibody [EPR1390Y]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
African Green Monkey Cell lysate - whole cell (COS-7 cells)
Gel Running Conditions
Reduced Denaturing (4-12% Bis-Tris)
Loading amount
100000 cells
Treatment
30 nM Bortezomib for 24 hours
Specification
COS-7 cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10%
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

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Verified customer

Submitted Jun 22 2015

Western blot abreview for Anti-Heme Oxygenase 1 antibody [EPR1390Y]

Good
Abreviews
Abreviews
abreview image
Application
Western blot
Loading amount
40 µg
Gel Running Conditions
Reduced Denaturing (10 % gel)
Sample
Rat Tissue lysate - whole (aorta)
Specification
aorta
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Oct 22 2013

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