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Chemi‐Lumi One Series for HRP

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Chemi-Lumi One is a series of high sensitive luminol-based chemiluminescence assay kits for Western Blotting. We offer three types of chemiluminescent substrates for Western blot detection with horseradish peroxidase enzyme (HRP).

Product Information

Product Name Chemi-Lumi One L Chemi-Lumi One Super Chemi-Lumi One Ultra
Product No. 07880 02230 11644
Usage General Use High Sensitive Ultra high Sensitive
Lower Detection Limit Low-picogram Mid-femtogram Low-femtogram
Required Working Solution Approx. 0.125 ml / cm2 Approx. 0.1 ml / cm2 Approx. 0.1 ml / cm2
Suggested Antibody Dilution Primary: 1:1,000-1:5,000
Secondary: 1:20,000-1:100,000
Primary: 1:1,000-1:20,000
Secondary: 1:20,000-1:200,000
Primary: 1:5,000-1:100,000
Secondary: 1:100,000-1:500,000
Reaction Period 1 min. 1 min. 5 min.
Comparable to ECL
SuperSignal Pico
ECL Prime
SuperSignal Dura
ECL Select
SuperSignal Femto
Sensitivity Sensitivity
<Condition>
Antigen: Anti-Mouse IgG (Goat), HRP Conjugated (Santa Cruz, sc-2005)
Detection: L (1 min. ), Super (1 min. ) and Ultra (5 min. )
Detector: LAS-3000 Super mode (Analyze 3 min. later after reaction with each substrate)
Expose time: 30 min.

Chemi-Lumi One L

  • Suitable for optimization approach of target proteins
  • Reasonable price in Chemi-Lumi One Series
  • Detect wide range of protein concentration

Comparison of sensitivity with competitors

Chemi-Lumi One L offers the similar sensitivity as T and W company's products and higher sensitivity than G company's products

Comparison of sensitivity

<Condition>

Gel 10% PAGE gel
Wash 0.1% t-TBS(1x), pH7.4
Blocking Blocking One
1st Ab Anti-β-Actin Mouse Monoclonal Antibody, (Santa Cruze sc-47778), 1:1,000
2nd Ab Anti-Mouse IgG (Goat), HRP Conjugated, (Santa Cruze sc-2005), 1:20,000
Detection period Chemi-Lumi One L, 1 min., Competitor G, T, W 1min.
Detector LAS-3000 High mode
Expose time 5 min.

Comparison of detection limit

Chemi-Lumi One L is suitable for detection of unknown protein concentration or optimization conditions.

Comparison of detection limit

<Condition>

Gel 10% PAGE gel
Wash 0.1% t-TBS(x1), pH7.4
Blocking Blocking One
1st Ab Anti-β-Actin Mouse Monoclonal Antibody, (Santa Cruze sc-47778), 1:1,000
2nd Ab Anti-Mouse IgG (Goat), HRP Conjugated, (Santa Cruze sc-2005), 1:20,000
Detection period Chemi-Lumi One L, 1 min.
Detector LAS-3000 High mode
Expose time 30 min.

References

  1. J Foo et al.: BMC complementary and alternative medicine 14.1 (2014)
  2. M Sato et al.: Scientific reports 4 (2014)
  3. M Tokumoto et al.: The Journal of toxicological sciences 38.6 (2013)
  4. H Hirakawa et al.: Biotechnology and bioengineering 110.7 (2013)
  5. H Yoshida et al.: Biochemical and biophysical research communications 418.2 (2012)
  6. H Fukuoka et al.: Growth Hormone & IGF Research 20.3 (2010)
  7. Q Yu et al.: Biochemical and biophysical research communications365.1 (2008)

Protocols

Chemi-Lumi One L pdf(PDF 100 KB)

Downloads

Chemi-Lumi One Series pdf(PDF 1.84 MB)

Chemi-Lumi One Super

  • Detects proteins at Mid-femtogram level with low background
  • Rapid substrate processing of blot
  • Applicable extention of exposure time and detectable lower expression level proteins

Comparison of sensitivity with competitors

Chemi-Lumi One Super is the most sensitive of competitors under the above consition, and its expore time is only 1 min., while others are required 5 min.

Comparison of sensitivity

<Condition>

Gel 10% PAGE gel
Wash 0.1% t-TBS, pH7.4
Blocking Blocking One
1st Ab Anti-β-Actin Mouse Monoclonal Antibody, (Santa Cruze sc-47778), 1:2,000
2nd Ab Anti-Mouse IgG (Goat), HRP Conjugated, (Santa Cruze sc-2005), 1:30,000
Detection period Chemi-Lumi One Super, 1 min.
Competitor G, T, W, B 5 min.
Detector LAS-3000 High mode
Expose time 5 min.

Comparison of background

Chemi-Lumi One Super performs under lower background, so users enable expose time to be longer for achieving more sensitive analysis.

Comparison of background

<Condition>

Gel 10% PAGE gel
Wash 0.05% t-TBS, pH7.4
Blocking Blocking One
1st Ab Anti-β-Actin Mouse Monoclonal Antibody, (Santa Cruze sc-47778), 1:1,000
2nd Ab Anti-Mouse IgG (Goat), HRP Conjugated, (Santa Cruze sc-2005), 1:10,000
Detection period Chemi-Lumi One Super, 1 min.
Competitor G, 5 min.
Detector LAS-3000 High mode
Expose time 1 min. and 10 min.


>> Comparison Data of Western Blotting Substrate with Chemi-Lumi One Super and competitors

References

  1. C Sakuma et al.: Molecular immunology63.2 (2015): 328-336

Protocols

Chemi-Lumi One L pdf(PDF 100 KB)

Downloads

Chemi-Lumi One Series pdf(PDF 1.84 MB)

Chemi-Lumi One Ultra

  • The most sensitive in Chemi-Lumi One Series
  • Longer signal duration
  • Applicable wider experimental condition due to low background

Comparison of sensitivity with competitors 1

Chemi-Lumi One Ultra is an extremely sensitive chemiluminescent substrate. Western blot of identical transferrin samples. The membranes were incubated with substrate that was prepared according to the manufacturers' instructions. The membranes were exposed to film for 2 minutes.

Comparison of sensitivity

Primary antibody : Rabbit anti-transferrin, 1: 30,000
Secondary antibody: Goat anti-rabbit IgG-HRP, 1: 200,000

Comparison of sensitivity with competitors 2

Chemi-Lumi One Ultra shows equal to or higher sensitivity compared to competitors'.

Application

<Condition>

Gel 10% PAGE gel
Wash 0.1% t-TBS(x1), pH7.4
Blocking Blocking One
1st Ab Anti-β-Actin Mouse Monoclonal Antibody, (Santa Cruze sc-47778), 1:300
2nd Ab Anti-Mouse IgG (Goat), HRP Conjugated, (Santa Cruze sc-2005), 1:1,000
Detection period 5 min.
Detector LAS-3000 High mode
Expose time 3 min.

Comparison of Abs concentration

Due to low background, Chemi-Lumi One Ultra is applicable to wide range of primary antibodies, and makes it easy to optimize condition.

Comparison of Abs concentration

<Condition>

Gel 10% PAGE gel
Wash 0.1% t-TBS(1x), pH7.4
Blocking: Blocking One
1st Ab Anti- PCNA Mouse Monoclonal Antibody, (Santa Cruze sc-56c)
2nd Ab Anti-Mouse IgG (Goat) HRP, (Santa Cruze sc-2005), 1:100,000
Detection period 5 min.
Detector: LAS-3000 High mode
Expose time 1:1,000, 3 min.
1:5,000, 5 min.
1:10,000, 8 min.

Protocols

Chemi-Lumi One L pdf(PDF 100 KB)

Downloads

Chemi-Lumi One Series pdf(PDF 1.84 MB)

Ordering Information

Product Name Storage Product No. PKG Size Price
Chemi-Lumi One L
Luminol 250 ml, Peroxide 250 ml
Sufficient substrate for 4,000 cm2 of blotting membrane
R 07880-70 1 kit e-Nacalai
Chemi-Lumi One Super
Luminol 50 ml, Peroxide 50 ml
Sufficient substrate for 1,000 cm2 of blotting membrane
R 02230-30 1 kit
Chemi‐Lumi One Ultra Luminol 50 ml, Peroxide 50 ml
Sufficient substrate for 1,000 cm2 of blotting membrane
R 11644-40 1 kit

[Patents]
Chemi-Lumi One Ultra is manufactured and patented by Cyanagen s.r.l and protected by US7803573; EP1962095; US7855287; EP1950207 US2012009603(A1); CA2742025; EP2405016 and foreign equivalents and other pending patents

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(Storage) RT: Room temperature, A: Cool and dark, R: Refrigerator, F: Freeze