The largest database of trusted experimental protocols

Image analysis system

Manufactured by Fujifilm
Sourced in Japan

The Image analysis system is a laboratory equipment designed to capture, process, and analyze digital images. It provides tools for tasks such as image acquisition, enhancement, measurement, and quantification. The system enables users to extract and interpret data from various types of images, supporting applications in fields like microscopy, medical imaging, and material science.

Automatically generated - may contain errors

10 protocols using image analysis system

1

Protein Extraction and Quantification

Check if the same lab product or an alternative is used in the 5 most similar protocols
After washed with PBS, cells were harvested and incubated with the RIPA buffer (20 mM Tris-HCl, pH 7.5, 150 mM NaCl, 1% Nonidet P-40, 1% sodium deoxycholate, 2.5 mM sodium pyrophosphate, 1 mM sodium vanadate, 1 μg/mL leupeptin, and 1mM Phenylmethanesulfonyl fluoride). After centrifugation at 20,000g for 20 min, the supernatant was collected and used as crude proteins. Protein concentration was determined using Bradford method (DC Protein Assay, Bio-Rad Laboratory, Hercules, CA, USA). Crude proteins (20 μg per lane) were separated by a 12.5% SDS-polyacrylamide gel and transferred onto a nitrocellulose membrane (Millipore, Bedford, MA). The membrane was blocked with 5% skimmed milk/PBS, incubated with primary antibodies (1000X-diluted), and then incubated with peroxidase-conjugated secondary antibodies. Signal development was conducted using ECL chemiluminescence reagent (Millipore) and the luminescent signals were acquired and semi-quantitated by an image analysis system (Fuji Film, Tokyo, Japan).
+ Open protocol
+ Expand
2

Western Blot Analysis of Protein Expression

Check if the same lab product or an alternative is used in the 5 most similar protocols
Western blotting was performed as previously described [18 (link)]. Briefly, cells were lysed in RIPA buffer containing a protease and phosphatase inhibitor cocktail (Sigma-Aldrich). The resulting crude proteins were separated using sodium dodecyl sulfate-polyacrylamide gel electrophoresis, transferred to polyvinylidene difluoride membranes (Immobilon; Merck), and reacted with first the primary antibodies and then the secondary antibodies. The bound antibodies were detected using enhanced chemiluminescence reagent (SuperSignal West Dura HRP detection kit; Pierce Biotechnology) and an image analysis system (Fujifilm). A densitometric analysis was performed for semiquantitation of the chemiluminescent signals.
+ Open protocol
+ Expand
3

Immunoblotting analysis of apoptosis-related proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols
Crude proteins were prepared using lysis buffer (1% Triton X-100, 0.1% SDS, 0.25 M sucrose, 1 mM EDTA, 30 mM Tris-HCl, pH 8.0) supplemented with protease inhibitor cocktail (Boehringer Mannheim, Indianapolis, IN), and subjected to a 12.5% SDS-polyacrylamide gel and transferred onto a nitrocellulose membrane as previously described [22 (link)]. The blot was subsequently incubated with 5% nonfat milk in PBS for 1 h, probed with a 1/1000 dilution of primary antibodies against Bcl-2, caspase-3, caspase-8, caspase-9, phospho-STAT-1 (pSTAT-1) and STAT-1 (Cell Signaling Technology, Beverly, MA, USA), or α-tubulin (α-TN, Santa Cruz Biotechnology, Santa Cruz, CA, USA) for 2 h, and then reacted with 1/2000 dilution of peroxidase-conjugated secondary antibody for 1 h. All incubations were carried out at 30°C, and intensive PBS washing was performed between each incubation. After the final PBS wash, the signal was developed by ECL chemiluminescence, and the relative photographic density was quantitated by image analysis system (Fuji Film, Tokyo, Japan).
+ Open protocol
+ Expand
4

Kidney Protein Extraction and Western Blot

Check if the same lab product or an alternative is used in the 5 most similar protocols
For western blot analysis, the kidneys were removed and snap-frozen in liquid nitrogen for protein extraction using RIPA (Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA). Next, 20 μg crude protein was subjected to 12.5% sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Following electrophoresis, the proteins were transferred onto a nitrocellulose membrane (GE-Healthcare, Little Chalfont, UK). The blots were blocked with 2% bovine serum albumin in phosphate-buffered saline (PBS), which was followed by 1 h incubation with 1:2,000 diluted primary antibodies against phosphorylated-p38 mitogen-activated protein kinase (p-p38 MAPK), phosphorylated-c-Jun N-terminal kinase (p-JNK) or NF-κB p65 (Santa Cruz Biotechnology, Inc.). Following washing with PBS, the reacted blots were incubated with peroxidase-conjugated secondary antibodies (BioSource International, Inc., Camarillo, CA, USA). Antigen-antibody complexes were identified by enhanced chemiluminescence (Pierce Biotechnology, Inc.) and the photographic density was quantified using an image analysis system (Fujifilm, Tokyo, Japan). GAPDH and lamin B1 were used as internal controls.
+ Open protocol
+ Expand
5

Western Blot Protein Analysis Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols
After treatment, the cells were detached by trypsinization, collected by centrifugation, and then lysed in the PBS lysis buffer containing protease and phosphatase inhibitor cocktail (Sigma-Aldrich). The cell lysates were centrifuged at 20,000 x g at 4 °C for 10 min, and then the supernatant was collected and used as a crude extract. The protein concentration in the extracts was assessed by Bradford protein assay (Bio-Rad Laboratories, Hercules, CA, USA). Extracts containing equal protein (30 µg) were separated by electrophoresis on an SDS-polyacrylamide gel. The electrophoresed proteins were transferred to a nitrocellulose membrane (PROTRAN BA85, 0.45 μm; Sigma) with a Transphor Unit (Bio-Rad Laboratories, Hercules, CA, USA). The transferred membrane was blocked with 1% (w/v) BSA in PBS followed by a 1-h incubation with the primary antibodies and then the secondary antibodies. The signal was developed with ECL chemiluminescence reagent (SuperSignal West Dura HRP Detection Kit; Pierce Biotechnology, Rockford, IL, USA), and the signals were acquired and quantitated with an image analysis system (Fujifilm, Tokyo, Japan).
+ Open protocol
+ Expand
6

Protease Expression Profiling Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols
Protease expression profiling analysis was performed as described previously.31 Briefly, the membranes coated with antibodies against multiple proteins were incubated with the mixture containing cell lysate. After the incubation, the membrane was reacted with streptavidin-HRP in array buffer and then incubated with the chemiluminescent reagent. The signals were analyzed by using an image analysis system (Fujifilm, Tokyo, Japan)
+ Open protocol
+ Expand
7

Western Blot Protein Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were lysed in RIPA buffer containing a protease and phosphatase inhibitor cocktail (Sigma-Aldrich) at 4 °C for 30 min, centrifuged at 20,000× g at 4 °C for 15 min to remove cell debris, and then the supernatant was used as crude protein extract. A protein assay was conducted using the Bradford method according to the manufacturer’s instruction (Bio-Rad Laboratories, Hercules, CA, USA). The crude proteins were separated using SDS-polyacrylamide gel electrophoresis and then transferred onto a polyvinylidene difluoride membrane (Immobilon, Merck, Billerica, MA, USA). The membrane was incubated with 3% (w/v) BSA in PBS for 1 h and then incubated with primary antibodies (1000-fold dilution) for 2 h. Thereafter, the membrane was washed with PBS containing 0.5% Tween-20 (PBST) and incubated with secondary antibodies (2000-fold dilution) for 2 h. The bound antibodies were detected using an ECL chemiluminescence reagent (SuperSignal West Dura HRP Detection Kit; Pierce Biotechnology, Rockford, IL, USA), and the resulting chemiluminescence signals were recorded and semi-quantitated with an image analysis system (Fujifilm, Tokyo, Japan). Signals from DMSO treatment were used as control.
+ Open protocol
+ Expand
8

Collagen I and NF-κB Western Blot

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were scraped off into culture medium and collected by centrifugation at 4000
g for 15 min before resuspension in the lysis buffer. Cell lysate
was subjected to a 12% SDS-PAGE gel and subsequently transferred onto a
nitrocellulose membrane as previously described. The resulting blot was incubated in
5% nonfat milk in PBS for 1 h and then probed with a primary antibody against
collagen type I and NF-κB p65, followed by reacting with an appropriate
peroxidase-conjugated secondary antibody for 1 h. Signals were detected using an
enhanced chemiluminescence detection kit (Amersham Biosciences, USA) and were
quantitated by an image analysis system (Fuji Film, Japan).
+ Open protocol
+ Expand
9

Western Blot Protein Analysis Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were lysed using RIPA buffer supplemented with protease and phosphatase inhibitor cocktail (Sigma-Aldrich) at 4 °C for 30 min. After centrifugation (20,000× g, 4 °C, 15 min) to remove insoluble debris, the supernatant (crude protein extracts) was collected for the following analysis. Protein concentration was assessed by Bradford method in accordance with the manufacturer’s instruction (Bio-Rad Laboratories, Hercules, CA, USA). Crude proteins were separated by SDS-polyacrylamide gel electrophoresis, transferred onto a polyvinylidene difluoride membrane (Immobilon, Merck-Millipore, Bedford, MA, USA), and then blocked with 3% (w/v) BSA/PBS at 25 °C for 1 h. Thereafter, the membrane was incubated with primary antibodies (1000-fold dilution) for 2 h and then incubated with secondary antibodies (2000-fold dilution) for 2 h after being washed with 0.5% Tween-20/PBS. The antibody–antigen complex was detected using an ECL chemiluminescence reagent (SuperSignal West Dura HRP Detection Kit; Pierce Biotechnology, Rockford, IL, USA), and the resulting signals were acquired and semi-quantitated using an image analysis system (Fujifilm, Tokyo, Japan). Signals from PBS treatment were used as control.
+ Open protocol
+ Expand
10

Western Blot Protein Detection Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were lysed in RIPA buffer containing a protease and phosphatase inhibitor cocktail (Sigma-Aldrich) at 4 °C for 30 min. The resulting lysates were centrifuged at 20,000× g at 4 °C for 10 min for the removal of insoluble debris, and then the supernatant was used for the protein assay using the Bradford method (Bio-Rad Laboratories, Hercules, CA, USA), and for electrophoresis using an SDS-polyacrylamide gel. The electrophoresed proteins were transferred to a polyvinylidene difluoride membrane (Immobilon-P, Millipore) and the membrane was blocked with 2% (w/v) skimmed milk in PBS. The blocked membrane was incubated with primary antibodies (1000-fold dilution) for 2 h, washed with PBS containing 0.5% Tween-20, and incubated with secondary antibodies (2000-fold dilution) for 2 h. The bound antibodies were detected using an ECL chemiluminescence reagent (SuperSignal West Dura HRP Detection Kit; Pierce Biotechnology, Rockford, IL, USA), and chemiluminescence signals were acquired and semi-quantitated with an image analysis system (Fujifilm, Tokyo, Japan). The results of the semi-quantitative analysis are presented as percentage of control (DMSO treatment).
+ Open protocol
+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!