Frozen liver tissues or cells were treated with protein extraction reagent supplemented with protease inhibitor cocktail (Bimake, B14002). Extracted protein samples were used for electrophoresis and then transferred onto polyvinylidene difluoride (PVDF) membrane. The membranes were blocked with 5%(w/v) skim milk (BD Bioscience, USA) for 1 h at room temperature, and then incubated with the following primary antibodies overnight at 4 °C: TWEAK (Abcam, ab37170, 1:1000), Tnfrsf12a(Abcam, ab109365, 1:1000), Phospho-RIPK1 (Cell Signaling Technology, 31122S, 1:1000), cleaved caspase-8 (Cell Signaling Technology, 8592T, 1:1000), cleaved caspase-3 (Cell Signaling Technology, 9664S, 1:1000), cleaved PARP (Cell Signaling Technology, 94885S,1:1000), RIPK3 (Cell Signaling Technology, 15828, 1:1000), MLKL (Abgent, AP14272B, 1:1000), GSDMD (Abcam, ab219800, 1:1000), GSDME (Abcam, ab215191, 1:1000), Tubulin (Sigma, T6199, 1:1000), GAPDH (Proteintech, 10494-1-AP, 1:5000). Original western blots are presented in Supplementary file.
T6199
Manufactured by Proteintech
Sourced in United States
The T6199 is a laboratory equipment product. It is used for [core function]. The product specifications and details are available on the Proteintech website.
Automatically generated - may contain errors
Lab products found in correlation
Ripk3, by Cell Signaling Technology (1 mentions)
Protease inhibitor cocktail, by Selleck Chemicals (1 mentions)
Phospho ripk1, by Cell Signaling Technology (1 mentions)
Ap14272b, by Abcam (1 mentions)
Cleaved caspase 8, by Cell Signaling Technology (1 mentions)
Cleaved parp, by Cell Signaling Technology (1 mentions)
Cleaved caspase 3, by Cell Signaling Technology (1 mentions)
Skim milk, by BD (1 mentions)
Gapdh, by Proteintech (1 mentions)
Ab3201, by Merck Group (1 mentions)
Mab1864, by GeneTex (1 mentions)
Gtx100911, by GeneTex (1 mentions)
T7451, by Merck Group (1 mentions)
Paraformaldehyde (pfa), by Electron Microscopy Sciences (1 mentions)
2 protocols using t6199
1
Comprehensive Necroptosis Protein Analysis
2
Immunocytochemistry of Cellular Microtubules
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Cells cultured in borosilicate glass Lab‐Tek eight‐well chambers (Nunc) were fixed in 4% paraformaldehyde (Electron Microscopy Sciences) at room temperature for 15 min. Fixed cells were permeabilized with 0.1% Triton X‐100 and then incubated in blocking solution (phosphate‐buffered saline with 2% bovine serum albumin) for 30 min at room temperature. To label cytosolic microtubules, primary ciliary membrane, and axonemal microtubules, cells were incubated for 1 h at room temperature with mouse antibody against α‐tubulin (1:500; Sigma Aldrich, T6199), rabbit antibody against Arl13b (1:500; Proteintech, 17711‐1‐AP), mouse antibody against glutamylated tubulin (1:100; Adipogen, AG‐20B‐0020‐C100), mouse antibody against acetylated tubulin (1:500; Sigma Aldrich, T7451), rat antibody against tyrosinated tubulin (1:100; Sigma Aldrich, MAB1864), rabbit antibody against cyclin B1 (1:500; Genetex, GTX100911), and mouse antibody against detyrosinated tubulin (1:100; MERCK, AB3201) each of which was diluted in blocking solution. Cells were then washed with PBS and incubated for 1 h with appropriate secondary antibodies (1:1,000 dilution; Thermo Fisher) at room temperature.
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