ECL Substrate Kit (Ultra High Sensitivity) (ab133409)
Overview
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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 bandDetection 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).
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Notes
Use:
- 100ml kit for 1000cm2 membrane
- 200ml kit for 2000cm2 membraneThe 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.
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Tested applications
Suitable for: WBmore details
Properties
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Storage instructions
Store at +4°C. Please refer to protocols. -
Components 200 ml 100 ml 20 ml Luminol/enhancer solution 1 x 100ml 1 x 50ml 1 x 10ml Peroxide Chemiluminescent Detection Reagent 1 x 100ml 1 x 50ml 1 x 10ml -
Research areas
Images
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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
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Western Blot 1/25000 primary antibody, 1/5000 secondary antibody, 10 second exposureEach 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
