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

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

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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 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.

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

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

Protein A purified
Clonality

Monoclonal
Clone number

EPR18161-128
Isotype

IgG
Research areas

Alternative Versions
- Anti-Heme Oxygenase 1 antibody [EPR18161-128] - BSA and Azide free (ab224677)
Compatible Secondaries
Immunohistochemistry reagents
- Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) (ab93684)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

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.

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

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)

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)

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)

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)

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)

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)

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 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 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)

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).

Datasheet

ab189491 has not yet been referenced specifically in any publications.

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

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