In the light of a previous delineation, the expression of Collagen-II was observed through ICC [30 (link)]. The treated HNPCs were collected and washed with PBS, and then fixed with 4% paraformaldehyde at room temperature for 15 min. Next, cells were treated with 0.2% of Triton X-100 (P0096, Beyotime, Shanghai, China) for 10 min, and incubated firstly with 10% goat serum (C0265, Beyotime, Shanghai, China) for 20 min, and then with the diluted primary antibody (Collagen II Monoclonal Antibody; MA1-37,493, Thermo Fisher Scientific, Waltham, Massachusetts, USA) at 4°C overnight. Subsequently, cells were incubated with the diluted secondary antibody (Rabbit Anti-Mouse IgG H&L; ab6728, Abcam, Cambridge, UK) at room temperature for 1 h. Later, the cells were treated with DAB working solution (P0202, Beyotime, Shanghai, China), and then washed with distilled water. Lastly, the Collagen-II content of cells was observed (under 200 × magnification) using fluorescence microscope (N-STORM, Nikon, Tokyo, Japan).
P0202
Manufactured by Beyotime
Sourced in China
The P0202 is a laboratory equipment designed for general use in scientific research and analysis. It serves as a device for measuring and analyzing various physical or chemical properties of samples. The core function of the P0202 is to provide accurate and reliable data to support scientific investigations and experiments.
Automatically generated - may contain errors
Lab products found in correlation
C0265, by Beyotime (1 mentions)
N storm, by Nikon (1 mentions)
Rabbit anti mouse igg h l, by Abcam (1 mentions)
Collagen 2 monoclonal antibody, by Thermo Fisher Scientific (1 mentions)
A0303, by Beyotime (1 mentions)
Ab6721, by Abcam (1 mentions)
Ab211737, by Abcam (1 mentions)
Ab276131, by Abcam (1 mentions)
Mwreg30, by BioLegend (1 mentions)
A0082, by Agilent Technologies (1 mentions)
Ab150160, by Abcam (1 mentions)
Ab6734, by Abcam (1 mentions)
F4 80 28463 1 ap, by Proteintech (1 mentions)
Dab substrate, by Beyotime (1 mentions)
Ab150077, by Abcam (1 mentions)
Image pro plus 6, by Media Cybernetics (1 mentions)
4 protocols using p0202
1
Collagen-II Expression in HNPCs
2
Immunohistochemical Analysis of Colon Tissue
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Paraffin‐embedded colon tissue sections (5 μm) were dewaxed, rehydrated, treated with 3% H2O2, heat‐bathed in citrate buffer for antigen retrieval, and blocked with normal goat serum. Subsequently, the sections were incubated with antibodies of mucin 3 (MUC3; 1:500, PAB031Mu01; Cloud‐Clone Corp.), Claudin‐1 (1:500, ab211737; Abcam Inc.), and tight junction protein 1 (ZO‐1, 1:500, ab276131; Abcam) overnight at 4°C, followed by incubation with goat anti‐rabbit IgG (1: 1,000, ab6721; Abcam) at 37°C for 20 min. The sections were then treated with horseradish peroxidase (HRP)‐labeled streptavidin‐working solution (A0303; Beyotime) and developed by DAB (P0202; Beyotime). Following counter‐staining with hematoxylin, the sections were observed under a microscope to calculate the number of positive cells in five randomly selected fields.
3
Histological Examination of Paraffin-Embedded Tissues
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The fixed tissue samples were embedded in paraffin. The paraffin blocks were sliced to a thickness of 4 μm. The EGTA (MVS-0098, Maxim biotechnologies, Fuzhou, China) was used for antigen repair. For histochemistry, F4/80 (28463-1-AP, Proteintech, Wuhan, China), MPO (SKU:023, Biocare Medical, Pacheco, CA, USA), VWF (A0082, DAKO, Glostrup, Denmark), and ITGB3 (MWReg30, BioLegend, San Diego, CA, USA) were used. The sections were incubated with the antibodies at 4 °C overnight. The sections were incubated with anti-rabbit IgG (#7074, Cell Signaling, Danvers, MA, USA) or anti-rat IgG (ab6734, Abcam, Cambridge, UK) for 1.5 h at room temperature and stained with DAB substrate (P0202, Beyotime, Shanghai, China). For immunofluorescence, the sections were incubated with ITGB3 (MWReg30, BioLegend, San Diego, CA, USA) overnight at 4 °C and then incubated with VWF (A0082, DAKO, Glostrup, Denmark) for 2 h at room temperature. For fluorescent staining, the sections were incubated with an Alexa 594-conjugated secondary anti-rat IgG antibody (ab150160, Abcam, Cambridge, UK) and an Alexa 488-conjugated secondary anti-rabbit IgG antibody (ab150077, Abcam, Cambridge, UK). The optical microscope and laser scanning confocal microscope were used to take images. The software ImageJ was used for quantifying the histochemistry staining.
4
Quantitative Analysis of FTO, Vimentin, and E-cadherin Expression in ESCC
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Tissue arrays were constructed using 28 pairs of human ESCC and corresponding paracancerous tissues, as well as nude mouse specimens. Paraffin-embedded sections and immunohistochemical staining were performed on all specimens to detect the expression of FTO for human ESCC and paracancerous tissues, and the expression of Vimentin and E-cadherin, for nude mouse specimens. In short, the slides were incubated overnight with the appropriate antibody at 4 °C and the secondary antibody at room temperature for 2 h. Then, Subsequent steps were performed using the immunohistochemical kit (Beyotime, FD008) and peroxidase (Beyotime, P0202). The intensity of staining was the staining intensity was detected using the IMAGE-PRO PLUS 6.0 software (Media Cybernetics, Silver Spring, MD, USA). Specific steps: (1) finding and measuring the section of interest; (2) adjusting the optical density; (3) acquiring, converting and saving the image; (4) correcting the background and background staining; (5) configuring the section of interest to determine the light density; (6) Checking the optical density. The positive staining in images was quantified as integral optical density (IOD)/area, that is, mean density = density sum/area sum [36 (link)].The relevant clinical information was provided in Table S1.
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!