Tumor tissues were fixed with 4% (v/v) paraformaldehyde before embedding them in paraffin. Tissue sections (4 μm) were later deparaffinized with xylene and rehydrated in a graded alcohol concentration series. Antigen epitope retrieval was induced by microwave heating. Tissue sections were then blocked for 1h, at room temperature, in goat serum blocking solution (Proteintech, B900780, China) and PBS (PBS). Tissue samples were incubated with primary antibody overnight at 4°C and the following day tissue the samples were stained using an anti-mouse/rabbit universal immunohistochemical detection kit (pk10006, proteintech) following the manufacturer’s instructions. Mounted sections were examined by light microscopy (Leica), and images were analyzed with Image-Pro Plus 448 (version 6.0).
B900780
Manufactured by Proteintech
Sourced in China
The B900780 is a laboratory equipment product. It is a centrifuge that is used to separate materials of different densities by applying centrifugal force. The centrifuge can be used to separate various biological samples, such as cells, proteins, or other macromolecules.
Automatically generated - may contain errors
Lab products found in correlation
Light microscopy, by Leica camera (2 mentions)
Anti mouse rabbit universal immunohistochemical detection kit pk10006, by Proteintech (2 mentions)
Bovine serum albumin (bsa), by Proteintech (1 mentions)
A1933, by Merck Group (1 mentions)
Influx flow cytometer, by BD (1 mentions)
Alexa fluor 488 conjugated goat anti mouse antibody, by Cell Signaling Technology (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
A16891, by ABclonal (1 mentions)
As039, by ABclonal (1 mentions)
4 protocols using b900780
1
Paraffin-Embedded Tissue Immunohistochemistry
2
Immune Cell Profiling in CSF and PBL
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All operations in this section were performed at low temperature. To evaluate PMN and MN populations, processed CSF cells or PBLs were stained with 1 μg/ml Hoechst 33342 for 5 min, then fixed in PBS containing 1% paraformaldehyde, and checked through an Influx flow cytometer (BD). For cell immunostaining, processed CSF cells or PBLs were blocked for 15 min in PBS containing 0.5% bovine serum albumin (Sigma-Aldrich, A1933) and 3% FBS and then incubated for 30 min with anti-VSIG4 (4 μg/ml, Santa Cruz, sc-53977), anti-CD1C (2 μg/ml, NOVUS, NBP2-62220), or equal concentrations of isotype-matched control antibodies. After washing three times with PBS containing 3% FBS, the suspension was incubated with 2 μg/ml Alexafluor 488-conjugated goat anti-mouse antibody (4408S, Cell Signaling) for 30 min in PBS containing 0.5% bovine serum albumin and 10% goat serum (B900780, Proteintech) and then incubated for another 5 min with 1 μg/ml Hoechst 33342. The cells were washed three times and fixed in PBS containing 1% paraformaldehyde. Cellular fluorescence was measured using an Influx flow cytometer, and data were analyzed using FlowJo software.
3
Immunohistochemical Analysis of CD68 in CRC
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Human CRC tumor tissues from the training cohort were fixed with 4% paraformaldehyde before being embedded in paraffin. Tissue sections (4 μm) were later deparaffinized with xylene and rehydrated with graded alcohol. Microwave heating was used to promote antigen epitope retrieval. Tissue slices were then blocked for 1 h at room temperature in goat serum blocking solution (Proteintech, B900780, China) and phosphate buffered saline (PBS). The sections were incubated overnight at 4°C with a primary antibody against CD68 (1:200; 76437S, CST). The next day, CRC tissue samples were stained according to the manufacturer’s instructions using an anti-mouse/rabbit universal immunohistochemical detection kit (pk10006, Proteintech) following the manufacturer’s instructions. Light microscopy (Leica) was used to examine mounted sections, and images were processed with Image-Pro Plus (version 6.0).
4
Quantifying COL1A1 expression in co-culture
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Protein expression of COL1A1 in each group was detected with confocal microscopy (LEICA TCS SP8) after coculture for 1 and 7 days. The samples were fixed with 4% paraformaldehyde, sealed with 10% goat serum (B900780, Proteintech, Wuhan, China), and then incubated overnight with primary antibody (COL1A1, A16891, ABclonal, Wuhan, China) 1:100. After rewarming at room temperature, the samples were incubated with secondary anti-rabbit IgG (AS039, ABclonal, Wuhan, China) 1:200 for 1 h. Subsequently, nuclei were stained using DAPI. Each sample randomly selected 3 vision, and each group of 3 samples, a total of 2 time nodes, measured 18 vision.
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