Nocodazole (M1404), PKH67 (green) (PKH67GL-1KT), PKH26 (red) (PKH26GL-1KT), brefeldin-A (B7651, used at 5 μg/ml), N-ethylmaleimide (E3876, used at 50 μM), LiCl2 (L-4408), PMA (used at 100 nM; P8139), and dynasore (D7693, used at 100 μM) were purchased from Sigma (St. Louis, MO, USA). TNF-α (210-TA) (used at 17 ng/ml), VEGF (used at 19 ng/ml; 293-VE), and neutralizing antibodies to VEGF (used at 1 μg/ml; MAB293) and VEGFR2 (used at 1 μg/ml; MAB3572) were purchased from R&D Systems (Minneapolis, MN, USA). Smac mimetic TL-32711 (A-1901) (used at 100 nM) was purchased from Active Biochem (Maplewood, NJ, USA). Cisplatin (NDC 0015-3220-22) was purchased from Bristol Laboratories (Princeton, NJ, USA). Antibodies to VEGF (for WB: Sc-7269), PKC-ζ (Sc-17781), K-Ras (Sc-30), and VEGFR2 (for WB: Sc-504) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Antibodies to E-cadherin (3195), p70S6K (2708), phospho-S6 ribosomal protein (Ser-235/236) (2211), S6 ribosomal protein (2317), syntaxin-6 (2869), and cdc42 (2466) were purchased from Cell Signaling Technology (Beverly, MA, USA). Antibody to β-catenin (610153) was purchased from BD Transduction Laboratories (Lexington, KY, USA). BI-D1870 (used at 5 μM; S2843) was purchased from Selleckchem (Houston, TX, USA).
Tl 32711 a 1901
Manufactured by Active Motif
Sourced in United States
The TL-32711 (A-1901) is a laboratory instrument used for digital imaging and analysis of biological samples. It is designed to capture and process high-quality images of various biological entities, such as cells, tissues, or molecular structures. The core function of this equipment is to provide users with a reliable and efficient tool for visualization, quantification, and data analysis in various life science applications.
Automatically generated - may contain errors
Lab products found in correlation
Tnf α 210 ta, by R&D Systems (3 mentions)
Mab293, by R&D Systems (2 mentions)
Bi d1870, by Selleck Chemicals (2 mentions)
Dynasore d7693, by Merck Group (2 mentions)
Cdc42 2466, by Cell Signaling Technology (2 mentions)
Antibody to β catenin 610153, by BD (2 mentions)
N ethylmaleimide e3876, by Merck Group (2 mentions)
Mab3572, by R&D Systems (2 mentions)
Brefeldin a b7651, by Merck Group (1 mentions)
A31604 02, by Thermo Fisher Scientific (1 mentions)
Vegfr2, by Santa Cruz Biotechnology (1 mentions)
Protein a sepharose cl 4b, by GE Healthcare (1 mentions)
Bi d1870 s2843, by Selleck Chemicals (1 mentions)
4 protocols using tl 32711 a 1901
1
Cellular Signaling Pathway Investigation
2
MLKL Silencing and Phosphorylation Assay in Cell Lines
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L-929 cells (CVCL_0462) were cultured in DMEM (12430-054; Gibco), 10% horse serum (HS; 26050-070; Gibco), 0.5 mM L-glutamine (25030-081; Gibco), and 1% penicillin/streptomycin (P/S; 15140-122; Gibco).
For mixed-lineage kinase domain-like (Mlkl) silencing, L929 cells were infected with lentivirus-containing shRNA against Mlkl (360819; Sigma Mission) at a multiplicity of infection (MOI) 100. Four days later, cells were harvested and protein extraction was performed to assess knock-down efficiency.
For inducing MLKL phosphorylation (p-MLKL), L929 cells were treated with 30-ng/ml human recombinant TNF-α (cyt-223-b; Prospec), 100 nM smac mimetic (TL-32711; A1901; Active Biochem), and 20 μM zVAD-fmk (V116; Sigma; TSZ) for 1 h, and then protein was extracted.
NIH 3T3 cells (CVCL_0594), were cultured in DMEM (12430-054; Gibco), 10% fetal bovine serum (FBS; A31604-02; Gibco), 0.5 mM L-glutamine (25030-081; Gibco), and 1% P/S (15140-122; Gibco). This cell line expresses high levels of MLKL and was used as positive control for testing anti-MLKL antibody.
For mixed-lineage kinase domain-like (Mlkl) silencing, L929 cells were infected with lentivirus-containing shRNA against Mlkl (360819; Sigma Mission) at a multiplicity of infection (MOI) 100. Four days later, cells were harvested and protein extraction was performed to assess knock-down efficiency.
For inducing MLKL phosphorylation (p-MLKL), L929 cells were treated with 30-ng/ml human recombinant TNF-α (cyt-223-b; Prospec), 100 nM smac mimetic (TL-32711; A1901; Active Biochem), and 20 μM zVAD-fmk (V116; Sigma; TSZ) for 1 h, and then protein was extracted.
NIH 3T3 cells (CVCL_0594), were cultured in DMEM (12430-054; Gibco), 10% fetal bovine serum (FBS; A31604-02; Gibco), 0.5 mM L-glutamine (25030-081; Gibco), and 1% P/S (15140-122; Gibco). This cell line expresses high levels of MLKL and was used as positive control for testing anti-MLKL antibody.
3
Comprehensive Cell Signaling Assay Protocol
4
Investigating Cell Death Signaling Pathways
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TL-32711 (A-1901; used at 100 nM throughout) was purchased from Active Biochem (Maplewood, NJ, USA), BI-D1870 (S2843) from Selleckchem (Houston, TX, USA), and TNF-α (210-TA; used at 17 ng/ml throughout) from R&D Systems (Minneapolis, MN, USA). LiCl2 (L-4408), H2O2 (1009) and DCF-DA (2′,7′-dichlorofluorescein diacetate: D6883) were purchased from Sigma (St. Louis, MO, USA). Cycloheximide (239763) was purchased from Calbiochem (San Diego, CA, USA), cisplatin (NDC 0015-3220-22) from Bristol Laboratories (Princeton, NJ, USA), and protein-A sepharose CL-4B (17-0780-01) from GE Healthcare (Pittsburgh, PA, USA). Antibodies to active caspase-3 (Sc-7148), caspase-9 (Sc-7885), PKC-ζ (Sc-17781), and p47phox (Sc-14015) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Antibody to caspase-8-Asp-391 (9496) was purchased from Cell Signaling Technology (Beverly, MA, USA).
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