ROTI®Lumin ultra, 2 x 20 ml in spray bottles
Transport temp. cooled
2 x 100 ml are sufficient for approx. 10,000 cm2 membrane (approx. 150 mini gel blots).
Excl. btw | 1 kit Per VE
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- Very easy to use
- Spray-and-go or mix by pipetting
- Superior performance for detections in lower femtogram range
- Perfectly suited for all exposition times
- Suitable for NC membranes, optimised for PVDF membranes
Significantly enhanced Western-Blot substrate based on luminol, for chemoluminescent detection of horseradish peroxidase (HRP) on membranes. Through a special, optimised composition, a particularly high signal strength is achieved for over 30 mins. with a second increase at 10 to 15 mins. after application. ROTI®Lumin ultra is, therefore, one of the most sensitive HRP-Chemoluminescence substrate available.
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. The superior signal strength allows detection of proteins in single femtograms, as well as significant reduction of concentrations of expensive antibodies.
- Spray the membrane with solution 1
- Afterwards spray membrane with solution 2
- Incubate approx. 1 min (wrap membrane in saran foil etc.)
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 ultra ready-to-use, for protein and cytoimmunochemistry
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 very short exposure we recommend use of ROTI®Lumin ultra.
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.
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.
20 ml (100 ml) solution 1 (Art. No. 3990) and 20 ml (100 ml) solution 2 (Art. No. 3991) in pump spray bottles.
Contents of this Kit may not be bought separately.
Superior sensitivity of ROTI®Lumin ultra even with a very short exposition of 10 seconds. Detection of 2 µl each of a HRP complex in dilution series 1:1 to 1:10.
a/A: ROTI®Lumin ultra; b/B: Product of a competitor, recommended for femtogram range.
a/b: 10 sec. exposition, A/B: 5 mins. exposition.
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.
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.
- Tussentotaal: 0.00
|3734.1||1 kit||plastic||2 x 20 ml in spray bottles||239,20 €||
Leveringsdatum onbekend op dit moment
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.
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.
|Solution 1||colourless to light yellow, clear|
|Light emission after 0, 5, 30 minutes (RLU)||complies|