The excised skin of mice was fixed in 10% formalin and embedded in paraffin. For histopathological examination, the tissue slices of 5-μm thickness were stained with hematoxylin and eosin (H&E) under the microscope (Olympus BX61, Tokyo, Japan). Epidermal thickness was calculated based on the means of four random sites of view per tissue and quantitated by ImageJ software as previously described (Liu X. et al., 2017 (link)). For immunohistochemistry, the tissue slices of 5-μm thickness were deparaffinized and incubated in hot citric acid buffer (PH 6.0) for antigen retrieval. After immediate cooling, slides were blocked with 5% BSA in TBST for 1 h and incubated with primary antibodies against ANGPTL4 (A2011, ABclonal) and Ki67 (GB13030-2, Servicebio) at 4°C overnight, followed by secondary antibodies at 1:200 for 30 min at room temperature. The immunostaining was visualized by 3,3-diaminobenzidine (Vector Laboratories, Burlington, CA, United States). Quantification of the DAB intensity in the images was performed using ImageJ software.
Gb13030 2
Manufactured by Wuhan Servicebio Technology
Sourced in China
GB13030-2 is a laboratory equipment product. It is designed for specific technical functions, but a detailed description cannot be provided while maintaining an unbiased and factual approach without extrapolation. Additional information about the core function and intended use is not available.
Automatically generated - may contain errors
Lab products found in correlation
Hrp conjugated secondary antibody, by Wuhan Servicebio Technology (2 mentions)
Gb23303, by Wuhan Servicebio Technology (2 mentions)
3 3 diaminobenzidine (dab), by Vector Laboratories (1 mentions)
Ki67 cell proliferation kit, by Sangon (1 mentions)
Ab70335, by Abcam (1 mentions)
Ab22604, by Abcam (1 mentions)
Dm6 b upright microscope, by Leica (1 mentions)
Tunel assay kit, by Abcam (1 mentions)
Anti lysozyme, by Abcam (1 mentions)
Anti f4 80, by Wuhan Servicebio Technology (1 mentions)
Anti cd3, by Cell Signaling Technology (1 mentions)
Yc0069, by Immunoway (1 mentions)
Hrp conjugated goat anti rabbit igg, by Wuhan Servicebio Technology (1 mentions)
Anti ki67, by Wuhan Servicebio Technology (1 mentions)
Anti n2icd, by Immunoway (1 mentions)
Hrp conjugated goat anti mouse igg, by Wuhan Servicebio Technology (1 mentions)
Gb11027, by Wuhan Servicebio Technology (1 mentions)
Gb111364, by Wuhan Servicebio Technology (1 mentions)
Ab7758, by Abcam (1 mentions)
Gb11022, by Wuhan Servicebio Technology (1 mentions)
11 protocols using gb13030 2
1
Histological Analysis of Murine Skin
2
Ki67 Immunostaining Protocol for Cell and Tumor Samples
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For cultured cells, Ki67 staining was carried out using a Ki67 Cell Proliferation Kit (Sangon Biotech, China) according to the manufacturer's recommendations. In brief, cells were rinsed with phosphate-buffered saline (PBS), fixed in 4% paraformaldehyde for 20 min, and permeabilized with 0.1% Triton X-100 in 0.1% sodium citrate for 10 min. After blocking with 10% FBS in PBS for 1 h, cells were incubated with a rabbit anti-Ki67 antibody (1:100, Sangon Biotech) overnight at 4°C followed by an incubation with a Cy3-conjugated goat anti-rabbit IgG (red, 1:100, Sangon Biotech) for 1 h. Nuclear were counterstained with DAPI (blue).
For tumors from mice model, Ki67 were detected by standard immunohistochemistry protocols. Slides were deparaffinized, hydrated and boiled in EDTA buffer (pH 9.0) for 2 min for antigen retrieval. After treated with 3% H2O2 for 25 min, slides were blocked with 3% BSA for 30 min, and then incubated with a rabbit anti-Ki67 antibody (1:100, GB13030-2, Servicebio, China) overnight at 4°C, followed by an incubation with HRP-conjugated secondary antibody (Servicebio) for 50 min. Then, the signal was developed by in DAB (brown) solution for 5 min and the nuclear were counterstained with hematoxylin (blue).
For tumors from mice model, Ki67 were detected by standard immunohistochemistry protocols. Slides were deparaffinized, hydrated and boiled in EDTA buffer (pH 9.0) for 2 min for antigen retrieval. After treated with 3% H2O2 for 25 min, slides were blocked with 3% BSA for 30 min, and then incubated with a rabbit anti-Ki67 antibody (1:100, GB13030-2, Servicebio, China) overnight at 4°C, followed by an incubation with HRP-conjugated secondary antibody (Servicebio) for 50 min. Then, the signal was developed by in DAB (brown) solution for 5 min and the nuclear were counterstained with hematoxylin (blue).
3
Immunohistochemical Analysis of Ki-67
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
4
Immunohistochemical Analysis of NONO, Ki67, and GPX1
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Sections (5 μm) were cut from paraffin-embedded tissues and incubated with primary antibody. Detection was performed through incubation with the species appropriate secondary antibody conjugated to horse radish peroxidase and the substrate DAB. The following primary antibodies were used: rabbit anti-NONO (ab70335, 1:200; Abcam; Waltham, MA, USA), rabbit anti-Ki67 (GB13030-2, 1:1,000; Servicebio; Wuhan, China) and rabbit anti-GPX1 (ab22604, 1:200; Abcam). Staining was evaluated independently to determine the histological score according to the proportion of positive staining cells and staining intensity.
5
Histological and Immunohistochemical Analysis of Colon Tissue
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
In brief, fresh colon tissues were fixed in 4% paraformaldehyde at 4 °C overnight, then subjected to paraffin imbedding sections. For H&E Staining, the sections were stained with haematoxylin and dehydration in graded alcohols and xylene. For immunolabeling, the sections were incubated with indicated primary antibodies: anti-CD3 [1:400, 85061; Cell Signaling]; anti-Ki67 [1:200, GB13030-2; Servicebio], anti-F4/80 [1:200, GB11027; Servicebio], anti-ChgA [1:400, ab45179; Cell Signaling] anti-CAI [1:200, SC39349; Santa Cruz], anti-Lysozyme [1:300, ab108502; Abcam], anti-N2ICD [1:200, YC0069; Immunoway], overnight at 4 °C in the dark. Then, the sections were incubated with either HRP–conjugated Goat anti-Rabbit IgG (1:200, G1215; Servicebio) or HRP–conjugated Goat anti-Mouse IgG (1:200, G1214; Servicebio) for 50 min at 25 °C. The subsequent detection was performed using the standard substrate detection of DAB. TUNEL assay kit was purchased from Abcam (ab66110). Images were taken by using Leica DM6 B Upright Microscope.
6
Histological Analysis of Tumor Samples
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
All the samples were obtained and fixed in 4% paraformaldehyde for 24 h. They were later embedded in paraffin. Histology slices were made on a rocking microtome. Hematoxylin and eosin staining was performed to observe the pathological dynamics in masses. To detect the cell proliferation in the tumor tissue, the sections were incubated with the Ki-67 antibody (1:500, GB13030-2, Servicebio, China) and detected using goat anti-rabbit IgG H&L (HRP) antibody (1:200, GB23303, Servicebio, China). DAB (DAKO, K5007, Japan) was used as the chromogen and then the section was counterstained with hematoxylin. The positive percentage calculated by four pathological professionals without prior knowledge of the experimental process. To detect the fibroblasts infiltration and blood vessels in the tumor tissue, the sections were incubated with collagen 1A1 (1:1,000, GB11022, Servicebio, China) and α-SMA (1:500, GB111364, Servicebio, China) antibody. The following procedure was the same with Ki67 staining. To detect the cell origin in the tumor, the sections were stained with anti-HLA G antibody (1:200, ab7758, Abcam) and detected with goat anti-mouse IgG H&L Alexa Fluor 488 (ab150113).
7
Sorafenib's Impact on IL-22 Signaling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Sorafenib (S1040, Selleck) was obtained from a reliable source and prepared as a stock solution. Recombinant human IL-22 (200-22, PeproTech) and antibodies for CD155 (ab267788, Abcam, UK), STAT3 (ab68153, Abcam, UK), p-STAT3(Y705) (ab267373, Abcam, UK), p-STAT3(S727) (ab219593, Abcam, UK), Ki-67 (GB13030-2, Servicebio, China), β-actin (ab6276, Abcam, UK) and GAPDH (ab8245, Abcam, UK) were obtained from reputable suppliers. Detailed information was available in Supplementary Table 1
8
Immunofluorescence Analysis of ZFP36L1 and Ki-67
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Each tissue section was sequentially deparaffinized, then rehydrated, and washed in phosphate-buffered saline (PBS). Subsequently, to unmask the antigen, tissue sections were microwaved in 10 mM sodium citrate buffer and washed with PBS. Sections were further incubated overnight with the primary antibodies toward ZFP36L1 (Proteintech; 12306-1-AP, 1:200) and Ki-67 (Servicebio; GB13030-2, 1:200) at 4°C, followed by goat anti-rabbit secondary antibody labeled with Alexa Fluor 488 at room temperature for 1 h. With the use of a fluorescence microscope (Olympus, Tokyo, Japan), signals were detected, and images were captured and analyzed using cellSens image analysis software (Olympus)
9
Immunohistochemical Analysis of Tumor Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For tumors from mice model, Ki67, p-Akt, and Cleaved Caspase3 were detected by standard immunohistochemistry protocols. Slides were deparaffinized, hydrated, and boiled in EDTA buffer (pH 9.0) for 2 min for antigen retrieval. After treated with 3% H2O2 for 25 min, slides were blocked with 3% BSA for 30 min, and then incubated with a rabbit antibody (1:100, GB13030-2, Servicebio, China) overnight at 4 °C, followed by an incubation with HRP-conjugated secondary antibody (Servicebio) for 50 min. Then, the signal was developed by in DAB (brown) solution for 5 min and the nuclear was counterstained with hematoxylin (blue).
10
Immunohistochemical Analysis of Tumor Tissue
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The tumor tissues isolated from sacrificed mice were immediately fixed in 4% paraformaldehyde for 24 h and embedded in paraffin. The embedded sections were sliced into 5-μm sections for staining. The deparaffinized and rehydrated sections were boiled in a high-pressure pot with sodium citrate antigen retrieval solution for 3 min. After three washes in PBS, the sections were incubated with 3% hydrogen peroxide in methanol for 15 min to inhibit endogenous peroxidase activity. After nonspecific reactions were blocked with 10% normal rabbit serum, the sections were incubated overnight at 4°C with rabbit polyclonal antibodies specific to Ki67 (GB13030-2, 1:200, Servicebio). Then, the sections were washed, incubated at room temperature for 50 min with horseradish peroxidase-conjugated secondary antibody (GB23303, 1:200, Servicebio), and counterstained with hematoxylin.
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!