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Chemiluminescent HRP Substrate (membrane) (ab5801)

Submit a review Q&A (4)References (2)

Overview

  • Product name

    Chemiluminescent HRP Substrate (membrane)

  • Product overview

    A 2 component chemiluminescent substrate for the detection of horseradish peroxidase (HRP) in western blotting.

     

    The Abcam Chemiluminescent HRP Substrate (membrane) is very sensitive and great care must be taken to optimize the individual assay components (antibodies, conjugates, solid phase, etc.) to minimize background reactivity associated with nonspecific immunochemical reactivities

  • Notes

    Prepare 1x substrate by mixing 1 ml of the chemiluminescent reagent with 2 ml of the chemiluminescent reaction buffer. Mix well and protect from light. More or less of the 1x substrate can be prepared using the same ratio.

    Stability: Stable for a minimum of 18 months when stored at 2-8ºC in the original container and protected from light. The 1x diluted reagent is stable for several hours at room temperature when protected from light.

  • Tested applications

    Suitable for: WBmore details

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab5801 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 at an assay dependent concentration.

Incubate membrane in chemiluminescent substrate for 1 minute then rinse off.
Notes
WB
Use at an assay dependent concentration.

Incubate membrane in chemiluminescent substrate for 1 minute then rinse off.

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

References (2)

Publishing research using ab5801? Please let us know so that we can cite the reference in this datasheet.

ab5801 has been referenced in 2 publications.

  • Wang P  et al. Decorin reduces hypertrophic scarring through inhibition of the TGF-ß1/Smad signaling pathway in a rat osteomyelitis model. Exp Ther Med 12:2102-2108 (2016). WB . PubMed: 27698699
  • Pacini S  et al. Constitutive expression of pluripotency-associated genes in mesodermal progenitor cells (MPCs). PLoS One 5:e9861 (2010). PubMed: 20360837

Customer reviews and Q&As

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1-4 of 4 Abreviews or Q&A

Question

Dear Sir/Madam Please let me know the difference between Chemiluminescent HRP Substrate (membrane) (ab5801) and ECL Western Blotting Substrate Kit (ab65623) because the customers do not know which one to choose for WB. Best regards.

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Answer

Thank you for your enquiry. I can confirm that both products (ab5801 and ab65623) can be used for Western blot application, however ab65623 can be applied for more test (50). Your customer may be interested in our very new range of WB reagents (Optiblot) and further information can found at this site: www.abcam.com/optiblot I am happy to arrange a discount if they would like to try this product. I hope this helps and if I can assist further, please do not hesitate to contact me.

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Question

Do you have protein extraction kits?  

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Answer

Thank you for calling Abcam. I have not been able to find a chemiluminescent EMSA kit in our catalogue. We do have several protein extraction kits though, please see the list below: Mammalian Cell Extraction Kit  ab65399 Plasma Membrane Protein Extraction Kit, ab65400 Please let me know if there is anything else I can help you with.

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Question

Thank you very much for reply. So do you supply the anti-pVIII protein of M13 phage antibody for ELISA (used in phage display experiment)? And HRP/AntiM13 and HRP substrate OPD (O-phenyldiamine dihydrochloride)?   Thank you very much.  

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Answer

Thank you for contacting us. We have two anti-M13 antibodies that may be of interest: ab6188 Anti-M13 + Fd Bacteriophage Coat Proteins antibody Binds specifically to phage coat proteins of M13 or fd phage. Click here (or use the following: https://www.abcam.com/M13--Fd-Bacteriophage-Coat-Proteins-antibody-ab6188.html). ab50370 Anti-M13 antibody [B62-FE2] (HRP) Binds to an epitope on pVIII (phage coat protein) covering the N-Terminal region of the g8p AEGDDPAKAAFDSLQASAT. Click here (or use the following: https://www.abcam.com/HRP-M13-antibody-B62-FE2-ab50370.html). We have a number of HRP kits, mainly for IHC. Here is a link to a HRP substrate than can be used for WB: ab5801 Chemiluminescent HRP Substrate (membrane) Click here (or use the following: https://www.abcam.com/Chemiluminescent-HRP-Substrate-membrane-ab5801.html). Please let me know if there is anything else I can help you with.

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Question

I am using your HMGB anitigen with antibody HMGB1 cat #ab12856 and your secondary ab6721 and getting a very low signal with traditional methods for HRP. I am look for a better signal but do not want to use film. Is the detection system ab5801 Ecl or Ecl+?

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Answer

I can confirm that the chemiluminescent peroxidase substrate ab5801 is actually a bit more sensitive than the ECL or ECL+ products. The fact you are having low signal with this substrate and the antibodies ab6721 and ab12856 could be due to a number of reasons, either protocol related or antibody/reagent related. The kit should be stable for a minimum of 18 months at 2-8 degrees C, if it is older than that it may have gone off. I would be happy to look at your protocol to see if the problem can be solved with protocol tips, if you can click on the link below this will take you to a short protocol questionnaire to put the information together. https://www.abcam.com/index.html?section=western&pageconfig=technical&intAbID=12856&mode=questionaire The antibody ab12856 has been tested in WB by a customer who left a review of their results, this was done on transfected cells and therefore maybe basal levels in your samples are very low? Do you have reports of your samples expressing GPCR MRGX1? We look forward to receiving your protocol to help you more,

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