• Product name
    ECL Substrate Kit (High Sensitivity)
    See all ECL Substrate kits
  • Product overview

    High Sensitivity ECL Substrate Kit ab133406 is designed for the detection of proteins with 23pg-187ng of protein per band. It uses an enhanced chemiluminescent substrate for western blotting that was developed for film imaging and is also compatible with CCD imaging. The high sensitivity ECL substrate produces a strong signal with very low background. Additionally, the ECL signal is long lasting, allowing repeated exposures without fear of losing data.

    Our ECL kits include our popular ECL Substrate Kit ab65623 and our high sensitivity ECL substrate kits:
    - this kit (High Sensitivity ECL Substrate Kit ab133406) to detect 23pg-187ng of protein per band
    - Very High Sensitivity ECL Substrate Kit ab133408 to detect 4.6pg-4.7ng of protein per band
    - Ultra High Sensitivity ECL Substrate Kit ab133409 to detect 1.2pg-2ng of protein per band 

    Detection ranges in pg and ng stated above should be used for guidance only as detection range is dependent on the molecular weight of a protein.

    This product was previously called Optiblot ECL Detect Kit (23pg-187ng).

  • Notes

    - 200ml kit for 2000cm2 membrane
    - 500ml kit for 5000cm2 membrane

    The primary antibody can often be diluted 5 to 10 fold more than usual when using this ECL substrate. A typical primary antibody dilution range using the substrate is 1/5000 - 1/20,000, with a typical secondary antibody dilution range of 1/20,000 - 1/100,000. Some optimisation may be required.


  • Tested applications
    Suitable for: WBmore details



Our Abpromise guarantee covers the use of ab133406 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.


  • Anti-Transferrin antibody (ab1223) at 1/25000 dilution. Lanes 1-14 Transferrin protein (ab91435), Loading dilution (Lanes 1-8): 12.5, 6.2, 3.1, 1.6, 0.8, 0.3, 0.19µg protein.



    HRP conjugated polyclonal to Rabbit IgG (ab97080) at 1/50000 developed using Optiblot ECL Detect and Vendor A ECL subtrate.

  • Each lane contains the following amount of COX2 recombinant protein (ab58868): 1) 100ng 2) 50ng 3) 25ng 4) 12.5ng 5) 6.25ng 6) 3.13ng 7) 1.56ng 8) 780pg 9) 390pg 10) 195pg 11) 98pg. The blot was probed with rabbit anti-COX2 antibody (ab15191) and with a rabbit secondary (ab97080). The blot was developed with 20ml of Optiblot ECL Detect.



This product has been referenced in:
  • Ozsoy AZ  et al. Altered expression of p97/Valosin containing protein and impaired autophagy in preeclamptic human placenta. Placenta 67:45-53 (2018). Read more (PubMed: 29941173) »
  • Kostarnoy AV  et al. Receptor Mincle promotes skin allergies and is capable of recognizing cholesterol sulfate. Proc Natl Acad Sci U S A 114:E2758-E2765 (2017). WB . Read more (PubMed: 28292894) »
See all 5 Publications for this product

Customer reviews and Q&As

1-6 of 6 Abreviews or Q&A

The solution is highly sensitive, generating signal very fast (beta actin came in 20-30 sec, using the optiblot ECL detection solution developed in chem doc instrument).

Abcam user community

Verified customer

Submitted Nov 09 2015

Visualization of Western blots

Good Good 4/5 (Ease of Use)
This abcam optiblot detect kit gives signal very fast like that of the west dura. For my protein of interest, Signal appeared between 50sec to 1.3mins in chemidoc, B-actin comes even faster (30-40sec). The concentration of the total protein loaded was 40ug).

Abcam user community

Verified customer

Submitted Aug 07 2015


ab133409 has been used for ELISA applications.

However, since there are many different readers, plates, etc., testing/evaluation is necessary to assess utility for your specific experimental conditions.

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Thank you for your enquiry.

I am sorry to confirm that ab133406 is not suitable for detection of AP. This detection system has been designed for optimal detection of HRP, and will not be as sensitive for detection of AP.

I am sorry I am not sure which application you are using, but the following two detection substrate kits are suitable for western blotting, and should also be suitable for IHC:

ab7413 Alkaline Phosphatase chromogen (BCIP/TNBT) https://www.abcam.com/index.html?datasheet=7413 https://www.abcam.com/index.html?datasheet=7413.

ab7468 Alkaline Phosphatase chromogen (BCIP/NBT) https://www.abcam.com/index.html?datasheet=7468 https://www.abcam.com/index.html?datasheet=7468.

I can recommend to review the online datasheets carefully for further information.

I hope this information will be helpful to you. If you have any further questions, please do not hesitate to contact us.

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Thank you for contacting us.

The alternative we have is ab133406; https://www.abcam.com/Optiblot-ECL-Detect-Kit-23pg-187ng-ab133406.html

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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Thank you for contacting us. I hope that you are well.

The ab133410 is a fluorescence based kit however the Roche kit cat # 11520709001 is a chemiluminescence based so this will not be a right alternative.

Please suggest ab133406, https://www.abcam.com/Optiblot-ECL-Detect-Kit-23pg-187ng-ab133406.html

This kit is vigorously tested and is fully guaranteed. Other alternatives are ab65623, ab65628, ab133408 and ab133409.

Please note that the kits ab133406 does not have secondary antibody as the Roche kit. However all HRP conjugated secondary would be suitable.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Free Rabbit monoclonal antibody with any purchase of a primary antibody, while stocks last! Quote “RABMAB-XBSMG” in your next primary antibody order. For more information, visit the following link: https://www.abcam.com/index.html?pageconfig=resource&rid=15447

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