Recombinant Anti-Heme Oxygenase 1 antibody [EPR18161-128] (ab189491)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR18161-128] to Heme Oxygenase 1
- Suitable for: WB, IHC-P, IP, ICC/IF, Flow Cyt (Intra)
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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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, IP, ICC/IF, Flow Cyt (Intra)more details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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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 Cyt (intra): HeLa and NIH/3T3 cells. IP: NIH/3T3 whole cell lysate.
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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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR18161-128 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Immunohistochemistry reagents
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
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 |
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WB | (1) |
1/2000. Predicted molecular weight: 33 kDa.
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IHC-P |
1/20000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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IP |
1/30.
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ICC/IF | (1) |
1/250.
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Flow Cyt (Intra) |
1/50.
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Notes |
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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. |
IP
1/30. |
ICC/IF
1/250. |
Flow Cyt (Intra)
1/50. |
Target
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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
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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
see all -
Alternative names
- 32 kD antibody
- bK286B10 antibody
- D8Wsu38e antibody
see all
Images
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All lanes : Anti-Heme Oxygenase 1 antibody [EPR18161-128] (ab189491) at 1/2000 dilution
Lane 1 : Wild-type A549 cell lysate
Lane 2 : HMOX1 knockout A549 cell lysate
Lane 3 : Mouse Spleen cell lysate
Lane 4 : HeLa cell lysate
Lane 5 : HEK-293 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Predicted band size: 33 kDa
Observed band size: 32 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-Heme Oxygenase 1 antibody [EPR18161-128] staining at 1/2000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab189491 was shown to bind specifically to Heme Oxygenase 1. A band was observed at 32 kDa in wild-type A549 cell lysates with no signal observed at this size in HMOX1 knockout cell line ab269503 (HMOX knockout A549 lysate ab259782).
To generate this image, wild-type and HMOX1 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % milk in TBS-0.1 % Tween®20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
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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).
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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 minutesBlocking: 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).
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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. -
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. -
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).
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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).
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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).
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Intracellular flow cytometric analysis of 90% methanol/ 4% paraformaldehyde fixed NIH/3T3 (mouse embryonic fibroblast) cell line labeling Heme Oxygenase1 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 Fluorr® 488, ab150077), at 1/2000 dilution was used as the secondary antibody.
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Intracellular flow cytometric analysis of 90% methanol/4% paraformaldehyde fixed HeLa (human cervix adenocarcinoma epithelial cell) cell line labeling Heme Oxygenase1 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 Fluorr® 488, ab150077), at 1/2000 dilution was used as the secondary antibody.
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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 Detection Reagent (HRP) (ab131366), was used for detection 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).
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (39)
ab189491 has been referenced in 39 publications.
- Jiang H et al. Sulfiredoxin-1 Inhibits PDGF-BB-Induced Vascular Smooth Muscle Cell Proliferation and Migration by Enhancing the Activation of Nrf2/ARE Signaling. Int Heart J 63:113-121 (2022). PubMed: 35034915
- Zhang M et al. Nrf2 Signaling Pathway Mediates the Protective Effects of Daphnetin Against D-Galactose Induced-Premature Ovarian Failure. Front Pharmacol 13:810524 (2022). PubMed: 35153783
- Shi YL et al. Trilobatin, a Natural Food Additive, Exerts Anti-Type 2 Diabetes Effect Mediated by Nrf2/ARE and IRS-1/GLUT2 Signaling Pathways. Front Pharmacol 13:828473 (2022). PubMed: 35153796
- Luo T et al. Protective Effect of Isoorientin on Oleic Acid-Induced Oxidative Damage and Steatosis in Rat Liver Cells. Front Pharmacol 13:818159 (2022). PubMed: 35185572
- Forcina GC et al. Ferroptosis regulation by the NGLY1/NFE2L1 pathway. Proc Natl Acad Sci U S A 119:e2118646119 (2022). PubMed: 35271393