Recombinant Anti-Heme Oxygenase 1 antibody [EPR1390Y] (ab68477)
![Western blot - Anti-Heme Oxygenase 1 antibody [EPR1390Y] (ab68477)](https://www.abcam.com/ps/products/68/ab68477/Images/ab68477-465254-anti-heme-oxygenase-1-antibody-epr1390y-western-blot-wildtype-a549-hmox1-knockout-a549-human-spleen-hl60.jpg "Western blot - 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
Related conjugates and formulations
Overview
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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.
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Positive control
- HepG2, A549, rat kidney, rat spleen, mouse kidney 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.
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
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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. 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
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
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Positive Controls
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Recombinant Protein
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Related Products
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).
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| IP |
1/20.
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| Notes |
|---|
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Flow Cyt (Intra)
Use at an assay dependent concentration. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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ICC/IF
1/100 - 1/250. |
|
WB
1/10000 - 1/50000. Detects a band of approximately 33 kDa (predicted molecular weight: 33 kDa). |
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IP
1/20. |
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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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 kDaLanes 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.
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![Immunocytochemistry/ Immunofluorescence - Anti-Heme Oxygenase 1 antibody [EPR1390Y] (ab68477)](https://www.abcam.com/ps/products/68/ab68477/Images/ab68477-267911-anti-heme-oxygenase-1-antibody-epr1390y-immunofluorescence.jpg)
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.
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![Western blot - Anti-Heme Oxygenase 1 antibody [EPR1390Y] (ab68477)](https://www.abcam.com/ps/products/68/ab68477/Images/ab68477-277343-anti-heme-oxygenase-1-antibody-epr1390y-western-blot.jpg)
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
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![Western blot - Anti-Heme Oxygenase 1 antibody [EPR1390Y] (ab68477)](https://www.abcam.com/ps/products/68/ab68477/Images/ab68477-341524-Ab68477.jpg)
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 secondsWe are unsure how to define the extra bands.
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![Flow Cytometry (Intracellular) - Anti-Heme Oxygenase 1 antibody [EPR1390Y] (ab68477)](https://www.abcam.com/ps/products/68/ab68477/Images/ab68477-276935-anti-heme-oxygenase-1-antibody-epr1390y-flow-cytometry.jpg)
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.
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![Immunoprecipitation - Anti-Heme Oxygenase 1 antibody [EPR1390Y] (ab68477)](https://www.abcam.com/ps/products/68/ab68477/Images/ab68477-267912-anti-heme-oxygenase-1-antibody-epr1390y-immunoprecipitation.jpg)
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.
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![Western blot - Anti-Heme Oxygenase 1 antibody [EPR1390Y] (ab68477)](https://www.abcam.com/ps/products/68/ab68477/Images/ab68477-267910-anti-heme-oxygenase-1-antibody-epr1390y-western-blot.jpg)
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 kDaBlocking and dilution buffer: 5% NFDM/TBST
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![Anti-Heme Oxygenase 1 antibody [EPR1390Y] (ab68477)](https://www.abcam.com/ps/products/68/ab68477/Images/ab68477-10-benefits-of-recombinant-antibodies.png)
Anti-Heme Oxygenase 1 antibody [EPR1390Y] (ab68477)
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (110)
ab68477 has been referenced in 110 publications.
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