Overview

  • Product name
    Anti-Heme Oxygenase 1 antibody
    See all Heme Oxygenase 1 primary antibodies
  • Description
    Rabbit polyclonal to Heme Oxygenase 1
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IHC-Fr, ICC/IF, Sandwich ELISA, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Dog, Human
  • Immunogen

    Recombinant rat HO-1 (Hsp32) lacking the membrane spanning region

  • Positive control
    • Recombinant Human and Rat HO-1 (Hsp32) Protein. IHC-P: FFPE mouse spleen normal and rat spleen normal.

Applications

Our Abpromise guarantee covers the use of ab13243 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. Detects a band of approximately 32 kDa (predicted molecular weight: 34.6 kDa).
IHC-Fr 1/200.
ICC/IF Use a concentration of 1 - 5 µg/ml.
Sandwich ELISA Use a concentration of 0.5 µg/ml. Can be paired for Sandwich ELISA with Mouse monoclonal [HO-1-1] to Heme Oxygenase 1 (ab13248). For sandwich ELISA, use this antibody as Detection at 0.5 µg/ml with Mouse monoclonal [HO-1-1] to Heme Oxygenase 1 (ab13248) as Capture.
IHC-P Use a concentration of 1 - 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

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.
  • Information by UniProt
  • Database links
  • 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

  • All lanes : Anti-Heme Oxygenase 1 antibody (ab13243) 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: 34.6 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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Heme Oxygenase 1 antibody (ab13243)

  • IHC image of Heme Oxygenase 1 staining in formalin fixed, paraffin embedded rat spleen normal tissue section*, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab13243, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

     

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • Anti-Heme Oxygenase 1 antibody (ab13243) (Rat microsomes)
    Predicted band size: 34.6 kDa

  • Alzheimer diseased section stained with ab13243.
  • ICC/IF image of ab13243 stained HepG2 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab13243, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • Standard Curve for Heme Oxygenase 1 (Analyte: Heme Oxygenase 1 protein (Tagged) (ab85243)); dilution range 1pg/ml to 1µg/ml using Capture Antibody Mouse monoclonal [HO-1-1] to Heme Oxygenase 1 (ab13248) at 5µg/ml and Detector Antibody Rabbit polyclonal to Heme Oxygenase 1 (ab13243) at 0.5µg/ml.

References

This product has been referenced in:
  • Merle NS  et al. Characterization of Renal Injury and Inflammation in an Experimental Model of Intravascular Hemolysis. Front Immunol 9:179 (2018). Read more (PubMed: 29545789) »
  • Xu X  et al. ERK1/2/mTOR/Stat3 pathway-mediated autophagy alleviates traumatic brain injury-induced acute lung injury. Biochim Biophys Acta 1864:1663-1674 (2018). WB . Read more (PubMed: 29466698) »
See all 66 Publications for this product

Customer reviews and Q&As

1-10 of 23 Abreviews or Q&A

Application
Western blot
Loading amount
25 µg
Gel Running Conditions
Reduced Denaturing
Sample
Human Cell lysate - whole cell (hepatocytes)
Specification
hepatocytes
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

Abcam user community

Verified customer

Submitted Dec 29 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Rat Tissue sections (Normal rat brain)
Specification
Normal rat brain
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.1% Triton X-100
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C

Abcam user community

Verified customer

Submitted May 10 2013

Answer

Vielen Dank für Ihre Anfrage.

Ich habe vier primäre AK für Sie herausgesucht, die alle in einer relativ hohen Verdünnung eingesetzt werden können, so dass sie für eine Weile reichen sollten. Das Sie alle Proben nun damit verarbeitet bekommen kann ich leider nicht versprechen, da die Antikörper ja auch in den meisten Fällen ein paar Optimierungsexperimente benötigen.

https://www.abcam.com/index.html?datasheet=9739 (or use the following: https://www.abcam.com/index.html?datasheet=9739).

https://www.abcam.com/index.html?datasheet=9969 (or use the following: https://www.abcam.com/index.html?datasheet=9969).

https://www.abcam.com/index.html?datasheet=13243 (or use the following: https://www.abcam.com/index.html?datasheet=13243).

https://www.abcam.com/index.html?datasheet=83339 (or use the following: https://www.abcam.com/index.html?datasheet=83339).


Als Sekundär-AK würde ich Ihnen den ab97069 Goat anti-Rabbit IgG H&L (HRP) secondary antibody empfehlen, da dieser bis zu einer Verdünnung von 1/50.000 eingestzt werden kann.



https://www.abcam.com/index.html?datasheet=97069 (or use the following: https://www.abcam.com/index.html?datasheet=97069).


Ich hoffe, dies hilft Ihnen weiter. Bitte zögern Sie nicht, sich wieder bei uns zu melden, falls Sie weitere technische Fragen haben. Falls Sie die Produkte Bestellen möchten oder mehr Informationen zB bezüglich des Preises wünschen, wenden Sie sich einfach an unseren Kundendienst.

Benutzen Sie unsere Produkte? Schicken Sie uns einen Abreview. Verdienen Sie sich eine Belohnung!
https://www.abcam.com/abreviews

Read More

Answer

Thank you for your reply.

I will then let the order through as it stands. We will endeavor to have the product with you as soon as possible but as I am sure you can understand we are reliant on delivery companies to move the package on our behalf. I will keep you updated with its progress.

As for the BCA kit and the extra BSA you have received compared to the 1 mL described. This is not unusual, we tend to supply a larger volume of all our products than we advertise to take into account any pipetting performed on our side. We want to make sure that the customer never receives less than expected and therefore typically over fill, with up to 10% of product. This should have no effect on the concentration of BSA supplied.

We do advise aliquotting the Anti-Heme Oxygenase 1 antibody when stored at -20°C regardless of the fact that it is in 50% glycerol. This is to avoid the antibody going through multiple freeze/thaw cycles (or more accurately with the glycerol added, temperature changes). This can affect the activity of the antibody and as a precautionary measure this is what we would advise. It also helps in that it can reduce the chance of contamination of the stock.

I hope this information has been of help. If I can be of any further assistance, please do not hesitate to ask.

Read More

Answer

Thank you for your reply.

We are indeed currently running an offer with our Cell Imaging book if you purchase any secondary antibody. Unfortunately I am unable to extend this offer to primary antibodies. I have had a look at the book and it is a very valuable resourcefor live cell imaging and light microscopy application. A review of the content can be found from the following link:

http://www.scionpublishing.com/shop/ProductImages/Cell%20Imaging%20Book%20Review.pdf

I think it is well worth getting if you are interested in this type of imaging. If your interested in the offer, please just quote the code CELL-TBVCQ with any secondary antibody order. More information on the offer can be found from the following link:

https://www.abcam.com/index.html?pageconfig=resource&rid=14705

In regards to a positive control tissue, we have found the protein to be highly expressed in liver and spleen tissue and so if you have access to this it may prove a useful control. Otherwise I would suggest having a look through the journals to see what other researchers have used in their experiments in conjunction to staining of Heme Oxygenase 1.

I hope this information has been of some help. If you have any further questions, please do not hesitate to contact us again.

Read More

Answer

I hope your grant application is going well. We have several antibodies against the targets you have mentioned which may be of interest to you:

HO-1

I would suggest looking at the rabbit polyclonal https://www.abcam.com/Heme-Oxygenase-1-antibody-ab13243.html. This has been shown to react with human, mouse and rat samples and has been used in ICC, IHC-Paraffin embedded, sELISA and Western blotting to date. We also have a very popular mouse monoclonal https://www.abcam.com/Heme-Oxygenase-1-antibody-HO-1-1-ab13248.htmlwhich has also been used with human, mouse and rat samples as well as in ELISA, Flow Cytometry, ICC,IHC-Frozen sections, and Western blotting. Bit I think you were more interested in antibodies raised in rabbit.

COX-2

For this target I would suggest having a look at the rabbit polyclonal https://www.abcam.com/COX2--Cyclooxygenase-2-antibody-ab15191.html. This has again been tested with mouse, rat and human samples and has been used in ICC/IF, IHC-Frozen sections, IHC-Parrafin embedded, IP, sELISA and Westernblotting.

We also have the NS-398 compound (COX-2 inhibitor) which is available under the catalogue number https://www.abcam.com/NS-398-ab120295.html.

I am afraid I have not been able to find a similar inhibitor of VEGF. Do you have a particular compound in mind? It may be that I can suggest we start stocking it.

I hope this information has been of help. If you have any further questions, please do not hesitate to ask.

Read More

Question
Answer

Thank you very much for your reply.

We are sending a free of charge vial of ab79854 on the order ***, which should arrive tomorrow. Please keep me updated about the results with this replacement, and let me know if there is anything else that we can do for you.

Have a nice day.

Read More

Answer

Thank you very much for your reply and for sending this additional information.

Would you be interested in tryingab79854or ab13248 free of charge? I can send either of these, if you would like. Alternatively I can issue a credit or refund.

I look forward to hearing from you. Please let me know if you have any questions or if there is anything else that we can do for you, and I'll be happy to help.

Read More

Question
Answer

Thank you again for your call earlier this week and for your patience while I have been in touch with the lab.

During the QC testing of this antibody, only a band at the expected molecular weight was seen in lysate samples.30ug of lysate was loaded per lane. The membrane was blocked with5% skim milk, and the primary antibody was diluted1:1000 in block and incubated with the membrane for 1 hour at room temperature. A goat anti-rabbit HRP-conjugated secondary antibodywas diluted 1:1000 and incubated with the membrane for 1 hour at room temperature. ECL substrate wasused for detection, and the membrane wasexposed for 20 seconds.

For our records, could you also tell me what dilution of ab13243 was used, and what blocking agent you used?

I've looked through several of the publications that used this antibody as well, and I haven't found any evidence of a specific band around 70 kDa. It does seem like this band may be non-specific. We do have other HO1 antibodies that I would be happy to send to you free of charge so you can compare the results. Either ab79854 and ab13248would be good to use in Western blot-

https://www.abcam.com/index.html?datasheet=79854&tab=abreviews&intabreviewid=20901

https://www.abcam.com/Heme-Oxygenase-1-antibody-HO-1-1-ab13248.html

If you would like to try one of these antibodies, please let me know your order or PO# and I will send it promptly.

I look forward to hearing from you, and if there is anything else that we can do for you please don't hesitate to ask. Have a nice day.

Read More

Question
Answer

Thank you so much for sending the images.

I will forward the lab's protocol and comments as soon as I have heard back from them. Please let me know if you need anything else in the meantime.

Read More

1-10 of 23 Abreviews or Q&A

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

Sign up