Immunohistochemical analysis of Bcl-2, caspase-3 and HSP70 were conducted in the testes using a UltraVision LP Large Volume Detection system (Thermo Fisher Scientific, Fremont, CA, USA) according to the manufacturer’s protocol. Briefly, paraffin sections of 5 μM thickness were immersed in a 10 mM citrate buffer solution (pH 6.0), heated in an autoclave sterilizer for 10 min, cooled for 30 min, and immersed in 3% aqueous hydrogen peroxide (H2O2) at room temperature for 30 min to block endogenous peroxidase. The sections were then washed in PBS, quenched with blocking buffer (Ultra V Block) and incubated overnight at 4 °C with the primary antibodies against Bcl-2, caspase-3 and HSP70. The sections were then washed and treated with horseradish peroxidase polymer for 15 min at room temperature. 3, 3′-diaminobenzidine (DAKO) was used for visualization of the immunoreaction. Finally, the sections were counterstained with haematoxylin. Negative control was performed by replacing the primary antibody with PBS. The slides were examined and photographed with a photomicroscope attached with a digital camera (Olympus, Tokyo, Japan).
Ultravision lp large volume detection system
Manufactured by Thermo Fisher Scientific
Sourced in United States
The UltraVision LP Large Volume Detection System is a laboratory equipment designed for the analysis of large volume samples. The system combines sensitive detection technologies to provide accurate and reliable results. Its core function is to enable the measurement and analysis of large volume samples in a laboratory setting.
Automatically generated - may contain errors
Lab products found in correlation
5 protocols using ultravision lp large volume detection system
1
Immunohistochemical Analysis of Testicular Markers
2
Immunohistochemical Analysis of GATA3 and STAT4 in Tissue Samples
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
IHC analysis of the transcription markers GATA3 and STAT4 was performed on formalin-fixed, paraffin-embedded samples using the UltraVision LP Large Volume Detection System (Thermo Fisher Scientific, Santa Clara, CA, USA). The slides were incubated with rabbit anti-STAT4 and GATA3 polyclonal antibodies at a 1:100 dilution. Indirect immunoperoxidase staining was performed according to the manufacturer’s instructions (Invitrogen, Thermo Fisher Scientific, Santa Clara, CA, USA). The sections were counterstained with Mayer’s hematoxylin, and the slides were observed under a microscope to check for positive reactions.
STAT4 and GATA3 immunostaining was scored on a semiquantitative scale by evaluating the intensity and percentage of positively stained cells on 10 high-power fields. The intensity of STAT4 and GATA3 staining was scored as follows: 0 = none, 1 = weak, 2 = moderate, and 3 = strong. The percentage of positively stained cells was assigned according to the following: 0 = 0%, 1 = 10%, 2 = 10–30%, and 3 = >30%.
STAT4 and GATA3 immunostaining was scored on a semiquantitative scale by evaluating the intensity and percentage of positively stained cells on 10 high-power fields. The intensity of STAT4 and GATA3 staining was scored as follows: 0 = none, 1 = weak, 2 = moderate, and 3 = strong. The percentage of positively stained cells was assigned according to the following: 0 = 0%, 1 = 10%, 2 = 10–30%, and 3 = >30%.
3
Immunohistochemical Analysis of Effusion Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For immunohistochemistry, five cases were selected for tuberculous pleural effusion and parapneumonic effusion, respectively, and four cases for each cell type of the malignant effusions. Immunohistochemical studies were conducted using the streptavidin-biotin-peroxidase method (UltraVision LP Large Volume Detection System; Thermo Fisher Scientific, Fremont, CA, USA). Briefly, sections (4 µm thick) were cut from paraffin-embedded material, deparaffinized with xylene, and rehydrated through a graded series of ethanol. After the inhibition of endogenous peroxidase, the sections were exposed to primary antibody at 4℃ overnight. Mouse monoclonal anti-human caspase-cleaved keratin 18 neoepitope M30 antibody (1:100, Peviva AB) was used as the primary antibody. The immunoreactive proteins were visualized with 3,3-diamino-benzidine, and the sections were counterstained with hematoxylin. For the negative controls, sections were incubated with nonimmunized mouse IgG instead of specific monoclonal antibodies and then processed according to the above procedure. Two pathologists evaluated the immunohistochemical staining patterns. First, the types of cells expressing M30 were determined. Thereafter, immunoreactivity for M30 was graded semiquantitatively as low, weak, moderate, or high.
4
Paraffin Embedding and IHC for SHMT2
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To obtain paraffin-embedded tissue blocks, the tissue samples were fixed in 10% formalin for 16 h at room temperature. Tissues were dehydrated, incubated in xylene, and embedded in paraffin. Paraffin-embedded tissue sections (4-μm-thick) were incubated at 56 °C for 3 h. Tissue sections were deparaffinized, hydrated, washed, and stained with H&E. For immunohistochemistry (IHC), tissue sections were deparaffinized, rehydrated, and washed, antigen retrieval was performed (10 mM citrate buffer, pH 6.0), and tissue was incubated with an anti-SHMT2 antibody (PA5-32228, 1:250, Invitrogen) for 30 min at room temperature. Detection was performed using the UltraVision LP large-volume detection system (Thermo Fisher Scientific, MA, USA).
5
Tumor Tissue Preparation and PD-L1 Detection
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
24 h after respective treatments, tumors were excised and first fixed in 5 L of formalin for 44 min. Subsequently, they were placed in an ethyl alcohol solution for 30 min before being transferred onto another ethyl alcohol solution at a higher concentration. This transfer was repeated 6 times. Following ethyl alcohol fixation, the samples were then placed in xylene solution at a low concentration for 45 min before being transferred onto another xylene solution at a higher concentration for a total of three times. Finally, the samples were then embedded in paraffin wax. The sample processing was performed using Leica Peloris (Buffalo Grove, IL, USA). The embedded tumor samples were cut into 3 μm slices with Leica RM2235 (Buffalo Grove, IL, USA) and incubated in hydrogen peroxide blocks for 10 min. The tumor-bound PD-L1 antibodies were then detected using the UltraVision LP Large Volume Detection System (Thermo Fisher, San Jose, CA, USA) according to the manufacturer’s guidelines.
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!