Overview

  • Product name
    Anti-Heme Oxygenase 1 antibody [HO-1-1]
    See all Heme Oxygenase 1 primary antibodies
  • Description
    Mouse monoclonal [HO-1-1] to Heme Oxygenase 1
  • Host species
    Mouse
  • Tested applications
    Suitable for: WB, ICC, Flow Cyt, ELISA, IP, Sandwich ELISA, IHC-P, ICC/IF, IHC-Frmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Cow, Dog, Human, Monkey
    Predicted to work with: Pig
  • Immunogen

    Synthetic peptide:

    MERPQPDSMPQDLSEALKEATKEVHTQAEN

    , corresponding to amino acids 1-30 of Human Heme Oxygenase 1.

  • Positive control
    • Recombinant Human or Rat HO-1 (Hsp32) Protein. HEK293 treated with 30 uM hemin for 18hrs (see Abreview). IHC-P: FFPE human spleen normal.

Properties

Applications

Our Abpromise guarantee covers the use of ab13248 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 Use a concentration of 4 µg/ml. Detects a band of approximately 32 kDa (predicted molecular weight: 34.6 kDa).
ICC 1/500.
Flow Cyt 1/100.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

ELISA Use a concentration of 2 µg/ml.
IP Use at an assay dependent concentration. PubMed: 24098580
Sandwich ELISA Use a concentration of 5 µg/ml. Can be paired for Sandwich ELISA with Rabbit polyclonal to Heme Oxygenase 1 (ab13243). For sandwich ELISA, use this antibody as Capture at 5 µg/ml with Rabbit polyclonal to Heme Oxygenase 1 (ab13243) as Detection.
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.
ICC/IF Use at an assay dependent concentration.
IHC-Fr 1/300.

Target

Images

  • All lanes : Anti-Heme Oxygenase 1 antibody [HO-1-1] (ab13248) at 1 µg/ml

    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 Mouse

    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

  • IHC image of Hem Oxygensae 1 staining in a section of formalin fixed, paraffin embedded normal human spleen 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 ab13248, 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.

    *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

  • The following proteins and lysates were electrophoresed; Lane 1 - Heme-Oxygenase-1 (Hsp32) Protein (50ng), lane 2 - Heme-Oxygenase-2 protein NSP-550 (100ng), lane 3 - MDBK Cell Lysate (20ug), lane 4 - Mouse liver microsome (20ug) and lane 5 - Dog liver microsome (20ug). ab13248 was applied at a concentration of 4ugml-1.
  • 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.
  • Lane 1 : MW marker
    Lanes 2-7 : Anti-Heme Oxygenase 1 antibody [HO-1-1] (ab13248)

    Lane 2 : Recombinant Rat Heme Oxygenase 1
    Lane 3 : Recombinant Human Heme Oxygenase 1
    Lane 4 : Recombinant Human Heme Oxygenase 2
    Lane 5 : MDBK cell lysate
    Lane 6 : Dog liver microsome
    Lane 7 : Mouse liver microsome

    Predicted band size: 34.6 kDa

  • Anti-Heme Oxygenase 1 antibody [HO-1-1] (ab13248) at 1/250 dilution + Human microsome lysate

    Predicted band size: 34.6 kDa

  • ab13248 staining Heme Oxygenase 1 in murine heart tissue by Immunohistochemistry (Frozen sections).
    Tissue was fixed in paraformaldehyde, permeabilized, blocked using 2% serum for 1 hour at 30°C and then incubated with ab13248 at a 1/300 dilution for 12 hours at 4°C. The secondary used was an Alexa-Fluor 488 conjugated goat polyclonal used at a 1/500 dilution.

    The green colour indicate Heme Oxygenase 1.
    The blue colour indicate nucleus stain DAPI.

    See Abreview

  • ab13248 at 10µg/ml staining Heme Oxygenase 1 in human lung cancer A2 cells by flow cytometery. The left image repersents staining with isotype control antibody and the right image show staining with ab13248.

References

This product has been referenced in:
  • Zhang Z  et al. Resolvin D1 promotes corneal epithelial wound healing and restoration of mechanical sensation in diabetic mice. Mol Vis 24:274-285 (2018). Read more (PubMed: 29643724) »
  • Liu Y  et al. Ameliorative effects of Antrodia cinnamomea polysaccharides against cyclophosphamide-induced immunosuppression related to Nrf2/HO-1 signaling in BALB/c mice. Int J Biol Macromol 116:8-15 (2018). WB ; Mouse . Read more (PubMed: 29730008) »
See all 92 Publications for this product

Customer reviews and Q&As

1-10 of 40 Abreviews or Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Spleen)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: pH 6.0 Citrate
Permeabilization
No
Specification
Spleen
Blocking step
PB ab64226 as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: RT°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Jan 23 2018

Application
Immunohistochemistry (Frozen sections)
Sample
Human Tissue sections (Skin)
Permeabilization
No
Specification
Skin
Blocking step
Serum as blocking agent for 1 hour(s) and 30 minute(s) · Concentration: 10% · Temperature: 21°C
Fixative
Acetone

Abcam user community

Verified customer

Submitted Jul 14 2016

Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing (8%)
Sample
Rat Cell lysate - whole cell (RAT AORTIC SMOOTH MUSCLE CELLS)
Specification
RAT AORTIC SMOOTH MUSCLE CELLS
Blocking step
Milk as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C

Mr. P Bingi

Verified customer

Submitted Oct 14 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (Glioblastoma primary cells)
Loading amount
30 µg
Specification
Glioblastoma primary cells
Gel Running Conditions
Reduced Denaturing (10%)
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Apr 16 2013

Question

1) Abcam product code ab 13248
2) Abcam order reference number or product batch number
3) Description of the problem : unable to detect bands in western blot
4) Sample preparation:
Type of sample (whole cell lysates, fraction, recombinant protein…): cell lysate
Lysis buffer 1x lysis buffer consisting of tris and SDS
Protease inhibitors: none
Phosphatase inhibitors none
Reducing agent none
Boiling for ≥5 min? yes/no
Protein loaded ug/lane or cells/lane 10-15ug perlane
Positive control rat spleen
Negative control none
5) Percentage of gel 8%
Type of membrane PVDF
Protein transfer verified
Blocking agent and concentration non-fat dry milk (5%), wash buffer, tween20 and distilled water
Blocking time atleast 1hr
Blocking temperature room temp.
6) Primary antibody (If more than one was used, describe in “additional notes”) : anti HO-1
Concentration or dilution 4ug/ml
Diluent buffer blocking buffer
Incubation time overnight
Incubation temperature: 40c
7) Secondary antibody: anti-mouse
Species:
Reacts against:
Concentration or dilution 1:5000
Diluent buffer blocking buffer
Incubation time 1hour
Incubation temperature: room temp.
Fluorochrome or enzyme conjugate: HRP linked
8) Washing after primary and secondary antibodies:
Buffer wash buffer (1x) with 0.1% tween20
Number of washes 3 times after primary and 5 times after secondary
9)Detection method
10) How many times have you run this staining? 3 times
Do you obtain the same results every time? yes
What steps have you altered to try and optimize the use of this antibody? none
Document attachment: Attaching images of your blot is strongly recommended and can greatly speed up our investigation of your problem.

Read More
Answer

Thank you for contacting us.

I have checked the details you have kindly provided. I would suggest the following troubleshooting steps;

- Please add cocktail of protease inhibitors in lysates.
- This antibody works perfect in reducing conditions. Please add DTT in sample buffer.
- Try heating the lysates at 100C for 10 minutes.
- Load 20-30ug of lysates.
- Try 5-10 ug/ml of antibody concentration.
- Please make sure the secondary is a anti-mouse antibody.

Please try these steps and let me know the results.

Many thanks for your cooperation. I will look forward to hearing from you soon.

Read More

Question
Answer

Thank you for calling us and for alerting us to the problem you are experiencing with our product. We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.

As we discussed on the phone, I am attaching our questionnaire so that we can gather further information regarding the samples tested and the protocol used. Once we receive the completed questionnaire, we will look at the protocol and see if there are any suggestions we can make that may improve the results. This information will also allow us to investigate this case internally and initiate additional testing where necessary. If the product was purchased in the last six months and is being used according to our Abpromise, we would be happy to replace or refund the antibody.

I look forward to receiving your reply.

Read More

Answer

Thank you for your email.

We have another kit, the Protein Quantitation Kit (Bradford Assay), ab102535, which may also be of interest to you. Or we also have Bradford reagent available from our catalogue, ab119216 and ab119216 for 500 and 1000 mL respectively. This can be used to set up your own protein quantification assay.

If you have any further questions relating to these products please do not hesitate to let me know.

Read More

Answer

Thank you for contacting Abcam regarding ab13248.


I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

Read More

Answer

Thank you for your reply.

Both the rabbit monoclonal ab52947 and the ab13243 have been shown to work well in IHC of paraffin embedded tissue sections. The results achieved can be seenfrom the images presentedon the datasheet of these products. Ab132436 has also been used in the following research article to stain mouse tissue:

Jortay J et al. Local induction of adiponectin reduces lipopolysaccharide-triggered skeletal muscle damage. Endocrinology 151:4840-51 (2010). PubMed: 20702578

Rabbit monoclonals are considered to be very good antibodies as they are specific and consistent like mouse monoclonals but also generally have very high affinity. However, I thought you wanted to use the antibody with rat tissue as well and as the rabbit monoclonal ab52947 has been shown not to react with rat samples this is why I did not suggest it to you.

We do not have either of the two compounds but I would be happy to pass these onto the lab to see if they are able to provide them. Do you know how much you would be interested in purchasing?

I look forward to receiving your reply.

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.

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1-10 of 40 Abreviews or Q&A

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