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AgriseraECL SuperBright (100 ml)

AS16 ECL-S-100 | extreme low femtogram detection

AgriseraECL SuperBright (100 ml) in the group Detection Reagents / Chemiluminescent detection at Agrisera AB (Antibodies for research) (AS16 ECL-S-100)

PRODUCT INFORMATION IN PDF

Qty: 
129 €

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product information
Background

AgriseraECL SuperBright for Western Blot detection is a high quality substrate for detection of horseradish peroxidase enzyme activity at extreme low femtogram levels offering low background and superior signal to noise ratio.

It is a ready to use 2 component system with low background and superior signal to noise ratios and highest sensitivity.

Host
Clonality
Clone
Purity
Format
Quantity 2 x 50 ml, two component ready to use solutions, enough for 50 midi blots (6.8 x 8.1 cm),  which is 2754 cm2
Reconstitution
Storage
  • 18 month shelf life at 2°C to 8°C.
  • Mixed working reagent is stable for several days at room temperature or at 4°C.
  • Exceptional lot to lot consistency.



Tested applications Western blot (WB)
Related products Low pico to mid femtogram detection:
AS16 ECL-N-10 | AgriseraECL Bright (10 ml, trial pack)
AS16 ECL-N-100 | AgriseraECL Bright (100 ml)
AS16 ECL-N-1L | AgriseraECL Bright (1L)
Extreme low femtogram detection:
AS16 ECL-S-10 | AgriseraECL SuperBright (10 ml, trial pack) 
AS16 ECL-S-1L | AgriseraECL SuperBright (1L)
Low pico to mid femtogram and extreme low femto detection in one set:
AS16 ECL-SN-10 | Agrisera ECL kit (Bright/SuperBright), 10 ml trial pack
AS16 ECL-SN | Agrisera ECL kit (Bright/SuperBright)
AS16 ECL-SN-1L, Agrisera ECL kit (Bright/SuperBright), 1L
Additional information This reagent offers extreme low femto level sensitivity combined with low background and superior signal to noise ratios.
application information
Recommended dilution
Expected | apparent MW
Confirmed reactivity
Predicted reactivity
Not reactive in
Additional information

User Instruction

  • Store reagents A and B in the darkness at 4-8°C.
  • Mix equal volumes of reagent A and B (chemiluminescent substrate) in a clean container and equlibrate to room temperature 30 minutes before use.
  • Prepare your membrane prior addition of chemiluminescent substrate, by a wash with a buffer used in your protocol (PBS or TBS or TBST-T). This will allow to remove any background prior to substrate contact.
  • Optimal visualization may be obtained up to 20 minutes after substrate contact. Incubation for 2 to 5 minutes can be enough.
  • Remove excess substrate by filter paper.
  • Cover blot with clear plastic wrap or sheet protector and expose either with x-ray film or CCD camera.
In some cases Tween can quench the reaction.
For best results clean containers and high quality water has to be used.

HS code for this product is: 3822.00.0002.
Selected references

Application example

Western blot detection using Agrisera ECLSuperBright reagent


10 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 5% non-fat milk dissolved in 20 mM Tris, 137 mM sodium chloride pH 7.6 with 0.1% (v/v) Tween-20 (TBS-T) for 1h at room temperature with agitation. Blots were incubated in Agrisera anti-RbcL primary antibody (AS03 037) at a dilution of 1: 20 000. 
The 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 following by washes and detection using Agrisera ECL SuperBright detection reagent. Images of the blots were obtained using a CCD imager (VersaDoc MP 4000) and Quantity One software (Bio-Rad). Exposure time was 60 seconds.

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

Caution:
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.



||| For other applications, usage on species other than stated above or any other questions, please use the LiveChat option or contact us at support@agrisera.com