For immunofluoresence staining, cells were seeded in confocal dish with or without naphplatin and CsA pretreatment for 24 h. The cells were fixed with 4% paraformaldehyde in PBS pH 7.4 for 10 min at room temperature and then permeabilizated with 0.5% Triton X-100 in PBS for 10 min. After being blocked with 2% BSA in PBS containing 0.1% Tween-20 for 30 min, cells were incubated overnight at 4 °C with Lamp1 (Abcam, ab25245, 1: 500), and Lamp2 (Abcam, ab25339, 1: 100), p65 (Cell Signaling Technology, #8242, 1: 400), in 2% BSA in PBS containing 0.1% Tween-20. After washing and incubating with secondary antibody for 1 h at room temperature, the DAPI (4',6-diamidino-2-phenylindole) (2 μg mL−1) was used to stain nucleus, which was then used for immunofluoresence analysis through OLYMPUS two-photon microscope. For immunohistochemical staining, melanoma tissues were isolated from tumour-bearing mice and fixed in 37% formalin and embedded in paraffin. Next, the Ultra Vision Quanto Detection System kit (Thermo Scientific) was used to incubate the section with the anti-Granzyme B (Abcam, ab4059, 1: 1,000) and observed by using the DAB Quanto kit (Thermo Scientific).
Ultravision quanto detection system kit
Manufactured by Thermo Fisher Scientific
Sourced in United States
The UltraVision Quanto Detection System kit is a laboratory equipment product designed for the detection and quantification of proteins in various biological samples. It provides a reliable and sensitive method for visualizing and measuring protein levels in a variety of applications.
Automatically generated - may contain errors
4 protocols using ultravision quanto detection system kit
1
Immunofluorescence and Immunohistochemistry Protocols
2
Histological Analysis of Olfactory Bulb Neurons
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The sample was fixed in formalin, then was dehydrated in 8 portions isopropyl-alcohol and embedded in cylindrical paraffin blocks 5 × 5 × 10 mm3 dimension, as for routine histological examination, and the block was measured via XPCT. After XPCT experiments, the samples were prepared for immunohistochemical analysis. Paraffin blocks were cut at 6-mm thickness sections. The sections were incubated with primary antibodies to -III-tubulin (dilution 1:500, Thermo Fisher Scientific, Waltham, MA, USA) and PGP9.5 (dilution 1:300, Thermo Fisher Scientific). The UltraVision Quanto Detection System kit by Thermo Fisher Scientific was used as the detection system. These sections were examined microscopically to identify areas within the blocks that contained histological features of interest – namely, neurons of different types forming layers of the OB. The Zeiss A1, made in Germany, which is a light compound microscope, was used for the microscopic measurements.
3
Immunofluorescence and Immunohistochemical Staining Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For immunofluoresence staining, cells were seeded in confocal dish for 24 h with or without CQ treatment and CsA pretreatment in some cases. The cells were fixed with 4% paraformaldehyde in PBS pH 7.4 for 10 min at room temperature and then permeabilizated with 0.5% Triton X-100 in PBS for 10 min. After blocking with 2% BSA in PBS containing 0.1% Tween-20 for 30 min, cells were incubated with NF-κBp65 (Cell Signaling Technology, #8242, 1 : 400), TFEB (Bethyl Laboratories, #A303-673A, 1 : 500), Lamp1 (Abcam, ab25245, 1 : 500), and Lamp2 (Abcam, ab25339, 1 : 100) in 2% BSA in PBS containing 0.1% Tween-20 overnight at 4 °C. After washing and staining with secondary antibody for 1 h at room temperature, nucleus were stained with DAPI (4',6-diamidino-2-phenylindole) (2 μg ml−1). The merged figures were analyzed by OLYMPUS two-photon microscope. For immunohistochemical staining, melanoma tissues were isolated from tumor-bearing mice and fixed in 37% formalin and embedded in paraffin. Next, the sections was incubated with the anti-Granzyme B (Abcam, ab4059, 1 : 1,000) by using the UltraVision Quanto Detection System kit (Thermo Scientific) and observed by using the DAB Quanto kit (Thermo Scientific).
4
Histological and Biochemical Analysis of Pancreas
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For the histological studies, the animals were sacrificed 30 days after the beginning of treatment. After anesthetization, the mice were disinfected on their ventral side with 75% ethanol15 . Body weight was recorded for each mouse before sacrifice and pancreas weight was recorded right after they were sacrificed. To study the biochemical effects of caerulein, gemcitabine and probiotics on blood cells and biochemistry, the blood was collected after sacrifice by cardiac puncture and immediately sent to Axel Biotechnology Inc., Taiwan for blood cell counting and biochemical analysis. Pancreas tissue was aseptically removed from the mice, and a small piece of tissue was fixed with 10% neutral buffered formalin (TONYAR Biotech. Inc. Taiwan) for 24–48 h at 4 °C. The fixed tissues were trimmed to an appropriate size, embedded in paraffin, sectioned and then stained with Hematoxylin and Eosin (H&E)16 ,17 . Vimentin (Vimentin (D21H3) XP Rabbit mAb, Cell Signaling Tech. # 5741) and Ki67 (Anti-Ki67 antibody KO tested, Abcam; ab15580) expression were detected using immunohistochemical staining methods in mouse pancreatic tissue. Immunohistochemical staining was performed using the UltraVision Quanto Detection System kit (Thermo Fisher Scientific Inc., Fremont, CA, USA) according to the manufacturer’s instructions18 .
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!