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ROTI®Lumin plus, 2 x 20 ml in spray bottles

ready-to-use, for protein and cytoimmunochemistry
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HRP substrate
Storage temp. +4 °C
Transport temp. cooled

Highly efficient chemoluminescence substrate for HRP-mediated Western-Blot detection systems.
2 x 100 ml is sufficient for approx. 10,000 cm2 membrane (approx. 150 mini gel blots).

€134.95/Pack Qty. 

excl. VAT. | 1 kit per Pack Qty.

Art. No. 3692.1

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Product details


  • Very easy to use
  • Spray-and-go or mix by pipetting
  • Optimised signal strength for detections in upper femtogram range
  • Well suited for medium- and long-term exposition
  • Suitable for NC membranes, optimised for PVDF membranes

Efficient Western-Blot substrate based on luminol, for chemoluminescent detection of horseradish peroxidase (HRP) on membranes. Through an optimised composition, high signal strength is achieved over a very stable plateau of over 30 mins.

By using pump spray bottles no time-consuming pre-mixing of solutions is required - substrate solution 1 and enhancer solution 2 are simply sprayed onto the membranes. Hereby, the spray technique guarantees a very smooth and regular distribution of the substrate on the membrane surface, hence avoiding artefacts caused by uneven distribution of pipetted substrate. Through the high signal strength, proteins in the upper femtogram range may be detected, while concentrations of the antibodies used may be reduced.

  • Spray the membrane with solution 1
  • Afterwards spray membrane with solution 2
  • Incubate approx. 1 min (wrap membrane in saran foil etc.)
  • Exposition

ROTI®Lumin is transformed by the HRP into an excited dianion in the presence of hydrogen peroxide. When the dianion is reversed back to its normal state, light is emitted. The light emission of ROTI®Lumin reaches its maximum within 5 minutes and remains at the maximum level for approx. 1-2 hours.



ROTI®Lumin plus ready-to-use, for protein and cytoimmunochemistry
Directions for use

For a mini blot of 7 × 8 cm size 3-4 pump strokes are necessary.
In case passed down protocols are to be used the spray-bottles may easily be unscrewed, mixing and applying the solutions by pipetting.
For medium or long-term exposure we recommend use of Roti®-Lumin plus.

3692_Diagramm_LichtemissionB
Figure:
Relative sensitivity of ROTI®Lumins. Measurement of the relative light emission following ECL reaction with solubilised HRP.
Competitor P: Product of a competitor, recommended for femtogram range.
3692_Diagramm_Lichtemission30min
Figure:
Typical course of light emission of the ROTI®Lumins: 0-30 mins., post mixing/spraying. Competitor P: Product of a competitor, recommended for femtogram range.

The kit contains

20 ml (100 ml) solution 1 (Art. No. 3936) and 20 ml (100 ml) solution 2 (Art. No. 3937) in pump spray bottles, or of 250 ml solution 1 (Art. No. 3936) and 250 ml solution 2 (Art. No. 3937) in refill-bottles.
Contents of this Kit may not be bought separately.

3692_RotiLuminPlus_Membranvergleich
Figure:
ROTI®Lumin plus has been optimised for use with ROTI®PVDF membranes (C). Detection of a polyclonal antibody (left to right 4 pg, 400 fg, 40 fg, 4 fg). Staining in PBST via a HRP-conjugated anti-rabbit antibody (1:2000). Exposure for 1 min.

A: ROTI®Nylon (Art. No. AE50.1);
B: ROTI®NC (Art. No. HP40.1);
C: ROTI®PVDF (Art. No. T830.1).
3692_Diagramm_Lichtemission5min
Figure:
Typical course of light emission of the ROTI®Lumins: 0-5 mins., post mixing/spraying. Competitor P: Product of a competitor, recommended for femtogram range.

RotiLumine_Sensitivitaet
Figure: Sensitivity of the ROTI®Lumine.
Sensitivity of ROTI®Lumin ultra is found in low femtogram range. Detection of a polyclonal antibody (left to right 4 pg, 400 fg, 40 fg, 4 fg) on ROTI®PVDF (Art. No. T830.1). Staining in PBST via a HRP-conjugated anti-rabbit antibody (1:2000).
a/A: ROTI®Lumin ultra; b/B: ROTI®Lumin plus; c/C: ROTI®Lumin; d/D: Product of a competitor, recommended for femtogram range. a/b/c/d: 10 sec. exposition, A/B/C/D: 1 min. exposition.


Technical Information
Enzyme Horseradish peroxidase 
Detection chemiluminescent 
Signal product Precipitate (Blot) 
ROTI®Lumin plus
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Downloads / MSDS

General information

Assay protocol for use of BCIP/NBT in immunoblot procedures:

Stock solutions (All solutions are stable for at least 1 year at 4 °C):
0.5 g NBT in 10 ml 70 % dimethylformamide;
0.5 g BCIP (p-toluidine salt) in 10 ml 100 % dimethylformamide.
Incubation buffer for alkaline phosphatase:
100 mM NaCl, 5 mM MgCl2,100 mM Tris (pH 9.5).
Fresh substrate solution: 66 μl NBT stock solution + 10 ml incubation buffer, mix well, add 33 μl BCIP stock solution. Use within 1 hour.
Blot development: Approx. 10 ml substrate solution per 15 x 15 cm2 membrane surface. Develop at room temperature until bands become visible (approx. 30 mins).
Reaction stop: Rinse with PBS/20 mM EDTA.


Detection of peroxidase activity (Immunoassay):

Dissolve 1 mg TMB in 0.1 ml dimethylsulfoxide (Art. No. 4720); add 9.9 ml of a 0.1 M sodium acetate solution (pH 6.0) (Powder: Art. No. 6779), filter and add H2O2 (Art. No. 8070) (final concentration 0.01 %).

Always prepare freshly!

Incubation 10-30 mins at room temperature (approx. 50 μl per microtitre well; finally add 50 μl 1 M H2SO4, Art. No. X873, per well).
Photometric quantitation at 450 nm.

Reference: Bos, E.S. et al., (1981) J. Immunoassay 2, (3/4), 187.


Certificates of Analysis

You can search for and download your certificate of analysis for the selected product here. Please provide your batch number.
The following analysis certificates have been found:

Type analysis

Solution 1colourless to light yellow, clear
Solution 2colourless
Light emission after 5 and 30 minutes (RLU)complies

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