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HomeLife ScienceProteomicsWestern Blot ECL Substrate Kit (Ultra High Sensitivity)
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ECL Substrate Kit (Ultra High Sensitivity)

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SKU: ab133409-20ml Categories: Life Science, Western Blot Tag: abcam
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  • Description
  • Properties
  • Application
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  • Datasheet
Description
  • 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.

Properties

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components200 ml100 ml20 ml
    Luminol/enhancer solution1 x 100ml1 x 50ml1 x 10ml
    Peroxide Chemiluminescent Detection Reagent1 x 100ml1 x 50ml1 x 10ml
Application

Application

The Abpromise guarantee

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.

ApplicationAbreviewsNotes
WB
Use at an assay dependent concentration.
Images

Images

  • Western Blot 1/50000 primary antibody, 1/5000 secondary antibody, 10 second exposure

    Western Blot 1/50000 primary antibody, 1/5000 secondary antibody, 10 second exposure

    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) at 1/50000 dilution and with a rabbit secondary (ab97080) at 1/5000 dilution. The blot was developed with 20ml of Optiblot ECL Ultra Detect.

    Exposure time: 10 seconds

  • Western Blot 1/25000 primary antibody, 1/5000 secondary antibody, 10 second exposure

    Western Blot 1/25000 primary antibody, 1/5000 secondary antibody, 10 second exposure

    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) at 1/25000 dilution and with a rabbit secondary (ab97080) at 1/5000 dilution. The blot was developed with 20ml of Optiblot ECL Ultra Detect.

    Exposure time: 10 seconds

Datasheet

Datasheet

ECL Substrate Kit (Ultra High Sensitivity)

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