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ATTO Products

Purpose and Application

  • Chemiluminescence imaging of Western blotting

  • Fluorescent imaging of fluorescent staining gels and fluorescent proteins (optional)

  • Bright-field imaging of CBB staining gels (optional)

  • Quantitative analysis of samples from the image (Concentration and Molecular weight)

  • Merge image of the chemiluminescent image and prestained marker of membrane image

 

Features

  • It is a new generation of chemiluminescence imaging devices that can be adapted to various applications such as western blotting, fluorescent gel photography, and CBB dyeing gel photography.

  • Cooling cameras are widely used to reduce noise during prolonged exposure, but high-performance CMOS cameras are rare and are not yet common. To address these challenges, ATTO developed a highly sensitive cooling CMOS camera using its technology.

  • The new "LuminoGraph I CMOS" offers advanced applications to meet a variety of research needs.

WSE-6710 LuminoGraph I CMOS 

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①CAMERA LENS FILTER
  
CAMERA : COOLING CMOS CAMERA
Pixels : 2688 x 1512 (4 Mpixels)
Lens : F0.95 single-focus lens
Filter (optional) : YA-3 (LPF560), ND-0.1 (diminished filter)
The high-sensitivity cooling CMOS camera enables high-sensitivity chemiluminescence samples.
  
If you use the sensitivity on gain function, 

"HQ" → "STD" → "HIGH" → "ULTRA"
You can increase the detection sensitivity without compromising the resolution.

②White light source (Optional)
CBB-dyed gel, silver-dyed gel, etc., are used with a white transmission light source with less uniform.
It can be photographed with a high-resolution image of

4 megapixels.
Photographed images can be stored in 16-bit TIFF and are suitable for quantitative analysis.

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③Cyan Transparent Light Source (Optional)
A transmission-type fluorescent irradiation device with a peak wavelength of 505 nm (half-width ± 25 nm) can detect various fluorescent staining reagents, including ethidium bromide.

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Unlike binning, gain-based sensitivity adjustment can increase the sensitivity itself without compromising image resolution.
  
It shows the results of western blotting of transferrin proteins that in a 1/2 dilution sequence from 10ng/lane with anti-transferrin antibodies. It is a photographic image of luminescence after reaction with an HRP luminescence substrate.

[Experimental conditions]
Gel : M-520L
Sample : Human Transferrin

                 (1 ng left to 1/2 dilution sequence from lane)
Transcription Criteria: QBlot W, 24V, 15 mins
Blocking : EzBlockCAS, 30 mins
Antibodies:
Primary antibody: anti-human transferrin rabbit polyclonal antibody
Secondary antibody : HRP-labeled anti-rabbit Ig antibody
emission detection : EzWestLumi plus
Detection device : LuminoGraph I CMOS
Exposure time : 1 min

A sample of HeLa cell extraction protein added 0-5 ng of human albumin per lane was separated by u-PAGEL H and transcribed by QBlot Mini, and the total protein on the PVDF membrane was detected by EzStain AQua MEM.
Western blotting was performed using anti-human albumin antibodies and light emission was detected by EzWestLumi plus.
The image above shows the total protein (bright field) and target protein (luminescence) taken with LuminoGtaph I CMOS.
The graph is the result of analyzing each image data with CSanalyzer 4 and normalizing the amount of albumin based on the signal value of all proteins.
We were able to detect albumin in a wide concentration range of 3 orders, ranging from 7 pg to 5000 pg, with a signal intensity approximate to the theoretical value.
LuminoGraph I CMOS can detect low concentrations of weak luminescence with high sensitivity, and can be photographed in a wide dynamic range without saturation of high concentrations of protein.

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image

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[Experimental conditions]
Gel : UH-T420
Sample : Cell extract with human albumin added
HeLa cell extract (20μg/all lanes)
human albumin
(5 ng from left / 1/3 dilution sequence from lane)
Transcription Criteria : QBlot M, 24V, 15 mins
Blocking : EzBlockCAS, 30 mins
ANTIBODY: HRP-LABELED ANTI-HUMAN ALBUMIN
emission detection : EzWestLumi plus
Photographic equipment : LuminoGraph I CMOS
Bright field : Interior lighting, 10 ms
Luminescence : 10s

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It shows the results of a 30-minute-stained gel Electrophoresis with a fluorescent reagent through a size marker of DNA.

[Experimental conditions]
Gel : EHR-T7.5L
Buffer : EzRun TG
Sample : Various DNA Molecular Weight Standards
Gel Dyeing : EzFluoroStain DNA
EzPreStain DNA & RNA
Photographic equipment : LuminoGraph I CMOS
filters : YA-3 (optional)
Light Source : CyanoView (optional)
Filming time : EzFluoroStain DNA 20 ms
EzPreStain DNA & RNA 50 ms

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image of a CBB dyed (EzStain AQua) gel taken using a combination of displayed light sources and filters.

The CBB stain gel was subjected to pseudo-color treatment with CS Analyzer after detection.

[Experimental conditions]
Gel : P-R16.5S (trisin gel)
Buffers: EzRun T
Sample : Chicken muscle extract, EzStandard II,
EzStandard LMW, EzProtein Ladder
Gel stain : EzStain AQua
Photographic equipment : LuminoGraph I CMOS
filters : ND-0.1 (optional)
light source : FLAT-Viewer (optional)
Exposure time : 300 ms

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