Anti-Heme Oxygenase 1抗体[HO-1-1] (ab13248)

概述

  • 产品名称
    Anti-Heme Oxygenase 1抗体[HO-1-1]
    参阅全部 Heme Oxygenase 1 一抗
  • 描述
    小鼠单克隆抗体[HO-1-1] to Heme Oxygenase 1
  • 经测试应用
    适用于: WB, ICC, Flow Cyt, ELISA, IP, Sandwich ELISA, IHC-P, ICC/IF, IHC-Frmore details
  • 种属反应性
    与反应: Mouse, Rat, Cow, Dog, Human, Monkey
    预测可用于: Pig
  • 免疫原

    Synthetic peptide:

    MERPQPDSMPQDLSEALKEATKEVHTQAEN

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

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

应用

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.

应用 Ab评论 说明
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.

靶标

Anti-Heme Oxygenase 1 antibody [HO-1-1] 图像

  • 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
    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



  • Predicted band size : 34.6 kDa
    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 1 : As above
    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.

Anti-Heme Oxygenase 1 antibody [HO-1-1] (ab13248)参考文献

This product has been referenced in:
  • Singh A  et al. Podophyllotoxin and Rutin Modulates Ionizing Radiation-Induced Oxidative Stress and Apoptotic Cell Death in Mice Bone Marrow and Spleen. Front Immunol 8:183 (2017). Read more (PubMed: 28289414) »
  • Arai A  et al. p62/SQSTM1 levels predicts radiotherapy resistance in hypopharyngeal carcinomas. Am J Cancer Res 7:881-891 (2017). WB ; Human . Read more (PubMed: 28469960) »

See all 43 Publications for this product

Product Wall

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
Username

Abcam user community

Verified customer

提交于 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
Username

Mr. P Bingi

Verified customer

提交于 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
Username

Abcam user community

Verified customer

提交于 Apr 16 2013

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 reduc...

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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 ph...

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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...

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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 th...

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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...

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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 http://www.abcam.com/Heme-Oxygenas...

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Thank you for sending us the additional information. This is very helpful.
I think your protocol looks fine. I'd like to ask a couple more questions, if you don't mind:
1) Are you using the same sample preparation protocol for all your sample...

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