Overview

  • Product name

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

    Ultra High Sensitivity ECL Substrate Kit ab133409 is a horseradish peroxidase (HRP) substrate specially developed for Western blotting detection of very low abundance proteins with 1.2pg-2ng of protein per band. ECL Substrate Kit ab133409 produces a strong, long-lived signal, which, combined with very low background levels, allows for long exposure times enabling the detection of low-abundance proteins. Additionally, the signal from ECL Substrate Kit ab133409 is linear with respect to protein amount over a broad range of concentrations, allowing the user to accurately quantify protein bands.


    ECL Substrate Kit ab133409 is most suitable for detection of low-abundance proteins or in situations when amounts of available primary antibodies are very limited and high dilution factors are desired, or when primary antibodies have relatively low binding constants.


    This ECL Substrate Kit also compatible with X-ray film detection, though the limited dynamic range of film will make resulting data less quantitative. The ECL substrate also produces a chemifluorescent signal that can be detected with a fluorescence imaging system using appropriate excitation and emission settings.


    Our ECL kits include our popular ECL Substrate Kit ab65623 and our high sensitivity ECL substrate kits:
    - 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
    - this kit (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 Ultra Detect Kit (1.2pg-2ng).

  • Notes

    Use:
    - 100ml kit for 1000cm2 membrane
    - 200ml kit for 2000cm2 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

Properties

Applications

Our Abpromise guarantee covers the use of ab133409 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.

Images

  • 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 Ultra Detect.

  • 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 Ultra Detect.

Protocols

References

This product has been referenced in:

  • Kalinina DS  et al. Age-Dependent Electrocorticogram Dynamics and Epileptogenic Responsiveness in Rats Subjected to Prenatal Hypoxia. Dev Neurosci N/A:1-11 (2019). Read more (PubMed: 30904914) »
  • Vasilev DS  et al. Transient Morphological Alterations in the Hippocampus After Pentylenetetrazole-Induced Seizures in Rats. Neurochem Res N/A:N/A (2018). Read more (PubMed: 29936569) »
See all 3 Publications for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Question
Answer

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.

Read More

Answer

Since you need a suitable ECL for imagers, I suggest to use ab133408 (Optiblot ECL Max Detect Kit (4.6pg-4.7ng)) which also corresponds with the ECL plus you are using at the moment.

ab133408 has been validated for CCD/laser use and generates a linear and quantitative signal that is very stable.

https://www.abcam.com/index.html?datasheet=133408 (or use the following: https://www.abcam.com/index.html?datasheet=133408).

Please note that stripping the membrane is not recommended.

ab133409 (Optiblot ECL Ultra Detect Kit (1.2pg-2ng)) is also suitable but is much more sensitive. This means that with average expression levels of proteins the dilution factor of the antibodies has to be newly optimised (higher dilution factor).

Unfortunately, we do not have samples sizes of our product since this is logistically not feasible with over 95000 products.

But I would like to confirm that our products are covered by our Abpromise for six month after purchase.

https://www.abcam.com/abpromise

Read More

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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