25 μg of RSO membranes were loaded onto each well of SDS-16% PAGE. After transfer onto the PVDF membrane, MelBSt proteins were detected with anti-His tag antibody, and imaged by the ImageQuantTM LAS 4000 Biomolecular Imager (GE Health Care Life Science)47 (link).
Imagequant las 4000 biomolecular imager
Manufactured by GE Healthcare
Sourced in United States, United Kingdom
The ImageQuant LAS 4000 is a biomolecular imager designed for the detection and quantification of a wide range of fluorescent and chemiluminescent signals. It utilizes a high-resolution CCD camera and advanced optics to capture detailed images of gels, membranes, and other samples.
Automatically generated - may contain errors
Lab products found in correlation
Pvdf membrane, by Merck Group (8 mentions)
Pierce bca protein assay kit, by Thermo Fisher Scientific (7 mentions)
Ripa buffer, by Thermo Fisher Scientific (6 mentions)
Protease inhibitor cocktail, by Roche (5 mentions)
Lumilight plus ecl substrate, by Roche (5 mentions)
Supersignal west femto maximum sensitivity substrate, by Thermo Fisher Scientific (4 mentions)
Horseradish peroxidase hrp conjugated secondary antibody, by Jackson ImmunoResearch (4 mentions)
Supersignal west pico chemiluminescent substrate, by Thermo Fisher Scientific (4 mentions)
Trans blot turbo transfer system, by Bio-Rad (4 mentions)
Bca protein assay kit, by Thermo Fisher Scientific (4 mentions)
Halt protease inhibitor cocktail, by Thermo Fisher Scientific (4 mentions)
β actin, by Cell Signaling Technology (4 mentions)
Western lighting chemiluminescence reagent plus, by PerkinElmer (3 mentions)
Laemmli sample buffer, by Bio-Rad (3 mentions)
Mini trans blot electrophoretic transfer cell, by Bio-Rad (3 mentions)
Ripa buffer, by Merck Group (3 mentions)
Ecl reagent, by Thermo Fisher Scientific (3 mentions)
Immobilon western chemiluminescent hrp substrate, by Merck Group (3 mentions)
β mercaptoethanol, by Bio-Rad (3 mentions)
Nitrocellulose membrane, by Thermo Fisher Scientific (3 mentions)
100 protocols using imagequant las 4000 biomolecular imager
1
Western Blot Analysis of MelB Protein
2
Immunoblotting of Leaf Protein Extracts
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For immunoblotting assays, leaf tissues collected at indicated time points/treatments were homogenised in protein extraction buffer [50mM Tris HCl (pH 8.0), 8M Urea, 50mM NaCl, 1% v/v NP-40, 0.5% Sodium deoxycholate, 0.1% SDS and 1mM EDTA] containing 20mM N-ethylmaleimide (NEM), 1X plant protease inhibitors cocktail (Sigma Aldrich) and 2% w/v Polyvinyl polypyrrolidone (PVPP). The homogenates were clarified by centrifugation, mixed with 2X Laemmli buffer (0.1M Tris pH 6.8, 20% glycerol, 4% SDS, 100mM DTT and 0.001% Bromophenol blue). Proteins extracts were boiled at 95°C and then separated onto Mini-PROTEAN ® TGX™ precast protein SDS-PAGE (4-15% gradient) gels (BIO-RAD, #4561083). Proteins were transferred onto polyvinylidene fluoride (PVDF) membrane by wet-transfer method. The membrane was blocked with 5% non-fat skim milk and immunoblots were performed with indicated primary antibodies [anti-SUMO1 (Abcam), anti-Actin C3 (Abiocode), anti-PR1 (Agrisera), anti-PR2 (Agrisera)] in 1X TBST at 4 0 C for overnight. Blots were washed thrice with 1X TBST and then incubated at RT for one hour with appropriate horse-radish peroxidase (HRP)conjugated secondary antibodies. The blots were then developed using ECL TM Prime western blotting system (GE Healthcare) and visualised in ImageQuant TM LAS 4000 biomolecular imager (GE Healthcare).
3
Western Blot Analysis of Protein Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
FZU-0038-056 or DMSO treated cancer cells were harvested using lysis buffer supplemented with a protease inhibitor cocktail (Roche Applied Science, Mannheim, Germany) for 30 min on ice. Proteins were collected and centrifuged at 4 °C, 13000 rpm for 10 min. Equal amounts of protein samples were then separated by SDS-PAGE electrophoresis and transferred to polyvinylidene fluoride (PVDF) membranes (Millipore, Bedford, MA). The membrane was blocked in 5 % skim milk for 1 hour, incubated with the primary antibody (1000 × dilution) overnight at 4 °C, washed 3 times with 1 × PBST for 10 min/time, and then incubated with secondary antibodies conjugated with horseradish peroxidase (HRP) (Jackson ImmunoResearch Laboratory, West Grove, PA) for 1 hour. They were then washed 3 times again with 1 × PBST. Finally, the membranes were incubated with Western Lighting Chemiluminescence Reagent Plus (PerkinElmer Life Sciences, Shelton, CT) and images were taken using an ImageQuant LAS4000 Biomolecular imager (GE Healthcare, UK).
4
Quantification of IL-38 in Murine Tregs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
IL‐38 expression in murine CD4+CD25+ Tregs was measured by Western blot following the manufacturer's instructions. In brief, nitrocellulose membranes were incubated at 4°C overnight with polyclonal rabbit antibodies against IL‐38 (1:1000) and monoclonal mouse antibody against β‐actin (1:1000), followed by incubation with horseradish peroxidase‐conjugated polyclonal goat anti‐rabbit secondary antibody (1:5000) or monoclonal rabbit antimouse antibody (1:5000) at room temperature for 1 hour. Finally, Western blot bands were visualized using ECLPlus (Amersham Biosciences) on an ImageQuant LAS 4000 biomolecular imager (GE Healthcare Life Sciences) and analysed using ImageJ software (US National Institutes of Health, https://imagej.nih.gov/ij/ ).
5
Hippocampus Protein Analysis in Rat Model
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
SD rats were classified into four groups (control, LIPUS, AlCl3, and LIPUS plus AlCl3), 4 h after the last LIPUS stimulation, six rats were killed and fresh hippocampus tissues were homogenized by T-Per extraction reagent (Pierce Biotechnology, Inc.). Lysates were centrifuged and supernatants were harvested and assayed with Protein Assay Reagent (Bio-Rad, CA, U.S.A.). A 40 μg protein was placed on 4–20% SDS-PAGE, transferred to Immun-Blot® PVDF membranes (Bio-Rad, CA, U.S.A.), blocked for 1 h with tris-buffered saline tween-20 (TBST)-containing 5% (w/v) skimmed milk, and then incubated overnight at 4°C in a solution with the primary antibody. A horseradish peroxidase-conjugated secondary antibody was incubated for 1 h at room temperature. Western Lightning ECL reagent Pro (Bio-Rad, CA, U.S.A.) was used to develop the signals, which was captured by ImageQuant™ LAS 4000 biomolecular imager (GE Healthcare Life Sciences, Pennsylvania, U.S.A.) and quantitated by Image J software (National Institute of Health, Bethesda, MD, U.S.A.).
6
Western Blot Detection of CREB5 and Viral Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
We performed SDS-PAGE and western blotting to detect the CREB5 and viral proteins. Total protein sample isolation and separation have been described elsewhere [25 (link)]. In brief, the cellular total proteins was boiled and separation in an 12% SDS-PAGE. The separated SDS-PAGE was then transferred to a polyvinylidene difluoride (PVDF) membrane (Bio-Rad Laboratories, Hercules, CA) and then washed. The membrane was and blocking with fast blocking solution (Biofuture Co., Taoyuan, Taipei) for 3 min prior to the primary and secondary antibodies incubation. The primary antibodies used were listed as the following: rabbit anti-EV-A71 VP (GeneTex, 1:1000), anti-Flag (Sigma, 1:3000 dilution) and anti-EV-A71 3D (1:5000 dilution); for detection of CREB5 (GeneTex, 1:2000) and β-actin (Sigma, 1:10000). The secondary antibodies were purchased from – (a) Goat anti-rabbit HRP secondary antibody (Abcam ab205718); (b) Goat anti-rabbit HRP secondary antibody (Abcam ab205719). The primary and secondary antibodies were diluted in the Super antibody mate solution (MDBio Co., Ltd., Taipei, Taiwan) followed by incubation ECL (BioMan biotech., Taipei, Taiwan) with the membranes. The signals of interested were detected using the ImageQuant™ LAS 4000 biomolecular imager (GE, USA).
7
Proteomic analysis with PLA2 antibodies
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Amicon Ultra-4 centrifugal filters (3kDa cut-off) came from Merck Millipore. Constituents of the Protease inhibitor cocktail [41 (link),57 (link)] were purchased from Sigma-Aldrich (St. Louis) and AMRESCO Inc. (Dallas, TX, USA). All electrophoresis grade chemicals used in two-dimensional gel electrophoresis (2DE) were purchased from AMRESCO Inc. 3-10NL IPG strips, tributyl phosphine and Polyvinylidene difluoride (PVDF) membrane (0.2 µM pore size) were from Bio-Rad (Hercules, CA, USA). Anti-human rabbit polyclonal iPLA2 antibody (PA5-27945) and anti-human mouse monoclonal sPLA2 antibody (ab-24498) were from Invitrogen and Abcam (Cambridge, UK), respectively. Blocking agents―non-fat dry milk powder and BSA―were from Devondale (Saputo Dairy, Australia) and Sigma Aldrich, respectively. HRP linked anti-mouse IgG (NA93IV) and anti-rabbit IgG (NA934VS) antibodies came from GE Healthcare (Buckinghamshire, UK). Lumunata Cresendo Western HRP substrate was purchased from Merck Millipore. The blots and gels were imaged using the Image Quant™ LAS 4000 Biomolecular Imager (GE Healthcare, Chicago, Il, USA).
8
Quantitative Western Blot Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total cellular proteins (10–30 μg) were extracted and chromatographed with sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) as described previously48 (link). After being transferred to a polyvinylidene difluoride (PVDF) membrane, the membrane was blocked in 1% bovine serum albumin or 10% nonfat milk for 1 h before incubation with specific primary antibodies. Secondary antibodies conjugated to horseradish peroxidase (HRP) were used to detect antigen–antibody complexes by using an enhanced chemiluminescence kit (Thermo Fisher Scientific Taiwan, Taipei, Taiwan) in an ImageQuant LAS 4000 Biomolecular Imager (GE Healthcare Life Sciences, Marlborough, MA).
9
Western Blot Analysis of EMT Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Preparation of cell lysates from SCC4 cells in 6-well plates and protein concentration was determined for each cell lysate using the BCA Protein Assay Kit (Thermo Fisher Scientific; Waltham, MA, USA). An amount of 30–50 μg of protein was loaded into each well containing 10% SDS-PAGE gel then transferred to polyvinylidene difluoride (PVDF) membranes (EMD Millipore; Temecula, CA, USA) for Western blot analysis, according to our previous research [32 (link),37 (link)]. Membranes were probed with their respective antibodies; anti-ILK (Santa Cruz; Dallas, TX, USA), anti-GSK3β(Santa Cruz; Dallas, TX, USA) anti-FAK (Cell Signaling; Danvers, MA, USA), anti-Akt (Cell Signaling; Danvers, MA, USA), anti-Snail (Santa Cruz; Dallas, TX, USA), anti-Twist (Santa Cruz; Dallas, TX, USA), anti-E-cadherin (Abcam; Cambridge, MA, USA), and anti-α-tubulin (Santa Cruz; Dallas, TX, USA). Immunoblot images were acquired using the ImageQuant™ LAS 4000 biomolecular imager (GE Healthcare Life Sciences; Pittsburgh, PA, USA).
10
Immunoprecipitation of ISL1 in Developing Retina
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
Revolutionizing how scientists
search and build protocols!