AgriseraECL Bright (100 ml)

85 €

AS16 ECL-N-100 


0 st
Item No:
AS16 ECL-N-100

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product information

AgriseraECL Bright for Western Blot detection is a high quality substrate for detection of horseradish peroxidase enzyme activity at a femtogram levels. Its a ready to use 2 component system with low background and superior signal to noise ratios.

Quantity 2 x 50 ml, two component ready to use solutions, enough for 50 midi blots (6.8 x 8.1 cm)

Stote in the dark at 2-8°C.
Highly stable for 18 month at 2°C to 8°C. Working stock is stable for 7 days at room temperature and 30 days at +4°C. Lot to lot consistency.

Tested applications

Western blot (WB)

Related products
Additional information

High sensitivity, low background and superior signal to noise ratios

HS code for this product is: 3822.00.0002

User Instruction

  • Store reagents A and B in the darkness at 4-8°C.
  • Mix equal volumes of reagent A and B in a clean container and equlibrate to room temperature 30 minutes before use.
  • Wash membrane with 0.2 M phosphate buffer pH 8.4 - 8.6 ading substrate.
  • Optimal results are obtained from 5 to 30 minutes after reaction is initiated.
application information
Recommended dilution
Expected | apparent MW
Confirmed reactivity
Predicted reactivity
Not reactive in
Additional information
Selected references

Application example

western blot using AgriseraECL Bright

1 µg of total protein from Arabidopsis thaliana leaf (1), Hordeum vulgare leaf (2) were extracted with Protein Extraction Buffer PEB (AS08 300). Samples were diluted with 1X sample buffer (NuPAGE LDS sample buffer (Invitrogen) supplemented with 50 mM DTT and heat at 70°C for 5 min and keept on ice before loading. Protein samples were separated on 4-12% Bolt Plus gels,  LDS-PAGE and blotted for 70 minutes to PVDF using tank transfer. Blots were blocked immediately following transfer in 2% blocking reagent (GE RPN 2125; Healthcare) for 1h at room temperature with agitation. Blots were incubated in the Agrisera anti-RbcL primary antibody (AS03 037) at a dilution 1: 20 000 (in blocking reagent) for 1h at room temperature. The primary antibody solution was decanted and the blot was rinsed briefly twice, and then washed 1x15 min and 3x5 min with TBS-T at room temperature with agitation. Blots were incubated in secondary antibody (goat anti-rabbit IgG horse radish peroxidase conjugated, recommended secondary antibody AS09 602, Agrisera) diluted to 1:20 000 in blocking reagent for 1h at room temperature with agitation. The blots were washed as above. The blot was developed for 5 min with AgriseraECL Bright (AS16 ECL-N-100). Images of the blots were obtained using a CCD imager (VersaDoc MP 4000) and Quantity One software (Bio-Rad). Exposure time was 5 minutes.

In the barley protein sample, a well characterized 44 kDa degradation product is observed (Kokobun et al. 2002).

Do not add any protein or HRP enzyme to reagent solution.
Optional: wash your tube, microtiter plate or membrane with 0.2 M solution of Sodium Phosphate, Dibasic, in Deionized Water. (Initial pH may be approximately 9) Bring the pH down to 8.4 with the slow addition of a 0.2M solution of Sodium Phosphate, Monobasic, in water.

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