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Triton x 100

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0.5% Triton X-100 is a non-ionic detergent commonly used in biochemical applications. It is a mild, non-denaturing detergent that solubilizes proteins and disrupts cell membranes without affecting the structure of proteins. This product is available in a 0.5% concentration.

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Lab products found in correlation

Alexa conjugated secondary antibody, by Abcam (2 mentions) Pe labeled anti cd206 antibody, by BioLegend (1 mentions) Facscan flow cytometer, by BD (1 mentions)

3 protocols using triton x 100

1

Quantifying RNA Polymerase II Distribution

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CHO cells were fixed with a buffer consisting of 4% formaldehyde (BioLegend) for 15 min at room temperature. Cells were then permeabilized with 0.5% Triton X-100 (BioLegend) for 15 min and treated with a blocking buffer consisting of 2% BSA in PBS for 4 h. To quantify distribution of RNA Poly II, we used an anti-RNA Poly II primary antibody. Detection of RNA Poly II was performed using Alexa-conjugated secondary antibodies (Abcam, Cat. No.150074). Cells were incubated with primary antibodies (diluted with 2% BSA buffer at a ratio of 1:100) overnight at 4 °C. After being washed to remove unbound primary antibodies, cells were incubated with the secondary antibody (diluted with 2% BSA buffer at a ratio of 1:1000) for 1 h under dark conditions. To quantify Poly II fluorescence, the images from the microscope were saved in the Tif format. Image J was used to invert the black and white color. The optical density of the inverted image was calibrated and set to scale. Thresholds were adjusted to get rid of auto-fluorescence background noise and fluorescence intensity in the area of interests was quantified.
Tajik A., Zhang Y., Wei F., Sun J., Jia Q., Zhou W., Singh R., Khanna N., Belmont A.S, & Wang N. (2016). Transcription upregulation via force-induced direct stretching of chromatin. Nature materials, 15(12), 1287-1296.
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2

THP1 Cells Phenotyping by Flow Cytometry

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THP1 cells were washed and re-suspended in PBS after PMA or PMA + IL-4 induction. To determine the expression of CD206, the cells were then incubated with PE-labeled anti-CD206 antibody (BioLegend, San Diego, CA, USA). To determine the intracellular expression of IL-10, IL-12, and TGF-β, the cells were fixed with 4% paraformaldehyde, permeabilized with 0.5% Triton X-100 (BioLegend), and stained with PE-labeled anti-IL-10, anti-IL-12, anti-TGF-β antibodies (BioLegend). After final wash, the labeled cells were analyzed by flow cytometry on a FACScan flow cytometer (BD Biosciences).
Cao H., Huang Y., Wang L., Wang H., Pang X., Li K., Dang W., Tang H., Wei L., Su M., Tang C, & Chen T. (2016). Leptin promotes migration and invasion of breast cancer cells by stimulating IL-8 production in M2 macrophages. Oncotarget, 7(40), 65441-65453.
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3

Quantifying RNA Polymerase II Distribution

Check if the same lab product or an alternative is used in the 5 most similar protocols
CHO cells were fixed with a buffer consisting of 4% formaldehyde (BioLegend) for 15 min at room temperature. Cells were then permeabilized with 0.5% Triton X-100 (BioLegend) for 15 min and treated with a blocking buffer consisting of 2% BSA in PBS for 4 h. To quantify distribution of RNA Poly II, we used an anti-RNA Poly II primary antibody. Detection of RNA Poly II was performed using Alexa-conjugated secondary antibodies (Abcam, Cat. No.150074). Cells were incubated with primary antibodies (diluted with 2% BSA buffer at a ratio of 1:100) overnight at 4 °C. After being washed to remove unbound primary antibodies, cells were incubated with the secondary antibody (diluted with 2% BSA buffer at a ratio of 1:1000) for 1 h under dark conditions. To quantify Poly II fluorescence, the images from the microscope were saved in the Tif format. Image J was used to invert the black and white color. The optical density of the inverted image was calibrated and set to scale. Thresholds were adjusted to get rid of auto-fluorescence background noise and fluorescence intensity in the area of interests was quantified.
Tajik A., Zhang Y., Wei F., Sun J., Jia Q., Zhou W., Singh R., Khanna N., Belmont A.S, & Wang N. (2016). Transcription upregulation via force-induced direct stretching of chromatin. Nature materials, 15(12), 1287-1296.
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