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

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: ICC/IF, WB, IP, Flow Cytmore details
Unsuitable for: IHC-P
Species reactivity

Reacts with: Mouse, Rat, Human
Immunogen

within Human Heme Oxygenase 1 aa 1-100 (N terminal). The exact sequence is proprietary.
Database link: P09601
Positive control
- HepG2, A549, rat kidney, rat spleen, mouse kidney 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
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.
This product is a recombinant rabbit monoclonal antibody.

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: 7.40
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, 0.05% BSA, 59% PBS
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR1390Y
Isotype

IgG
Research areas

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
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.
Flow Cyt		Use at an assay dependent concentration. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Application notes

Is unsuitable for IHC-P.

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

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

Hek293 & HL60 presumed negative or very low expression.

Loading control GAPDH at 38kDa
Immunocytochemistry/ Immunofluorescence - Anti-Heme Oxygenase 1 antibody [EPR1390Y] (ab68477)This image is courtesy of an anonymous Abreview

ab68477 (unpurified) staining Heme Oxygenase 1 in mouse embryonic fibroblasts by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with formaldehyde, permeabilized with 0.2% Triton X-100 and blocked with 1.5% serum for 10 minutes at 25°C. Samples were incubated with primary antibody (1/100 in 1X HBSS + 0.02% Triton X-100 + 1.5% FBS) for 3 hours at 25°C. An Alexa Fluor^® 594-conjugated goat anti-rabbit IgG polyclonal (1/1000) was used as the secondary antibody.
See Abreview
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.
Flow Cytometry - Anti-Heme Oxygenase 1 antibody [EPR1390Y] (ab68477)

Flow cytometry analysis of A549 (human lung carcinoma) cells labeling with purified ab68477 at 1/200 dilution ( 1ug/ml) (Red). Cells were fixed with 4% paraformaldehyde and 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)

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)

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
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 : HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate
Lane 2 : A549 (Human lung carcinoma cell line) whole cell lysate

Lysates/proteins at 15 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 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
Western blot - Anti-Heme Oxygenase 1 antibody [EPR1390Y] (ab68477)

All lanes : Anti-Heme Oxygenase 1 antibody [EPR1390Y] (ab68477) at 1/2000 dilution (purified)

Lane 1 : HepG2 cell lysate
Lane 2 : A549 cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : HRP conjugated Goat anti-Rabbit at 1/2000 dilution

Predicted band size: 33 kDa
Observed band size: 33 kDa
Additional bands at: 125-130 kDa, 30 kDa, 48 kDa, 52 kDa. We are unsure as to the identity of these extra bands.

Datasheet

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

This product has been referenced in:

Liu S et al. Gastrodin protects MC3T3-E1 osteoblasts from dexamethasone-induced cellular dysfunction and promotes bone formation via induction of the NRF2 signaling pathway. Int J Mol Med 41:2059-2069 (2018). Read more (PubMed: 29393365) »
Feng L et al. Icariside II alleviates oxygen-glucose deprivation and reoxygenation-induced PC12 cell oxidative injury by activating Nrf2/SIRT3 signaling pathway. Biomed Pharmacother 103:9-17 (2018). Read more (PubMed: 29635133) »

See all 19 Publications for this product

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

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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

Abcam user community

Verified customer

Submitted May 13 2016

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%

Abcam user community

Verified customer

Submitted Jun 22 2015

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

Abcam user community

Verified customer

Submitted Oct 22 2013

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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

Overview

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Datasheets and documents

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References

This product has been referenced in:

Customer reviews and Q&As

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

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

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