The largest database of trusted experimental protocols

3 protocols using tumor necrosis factor (tnf)

1

Rat Bone Marrow-Derived Macrophage Culture

Check if the same lab product or an alternative is used in the 5 most similar protocols
Primary BMDMs were obtained from rats’ rear leg’s bone marrow and differentiated to macrophages for 6 days in DMEM medium supplemented with 10% heat-inactivated FBS, 1% non-essential amino acids, penicillin, streptomycin, 2 mM glutamine, and 25 ng/ml M-CSF at 37 °C with 5% CO2.
Human recombinant TNF, Nec-1, zVAD, and BV6 were all synthesized at Genentech. LPS was purchased from InvivoGen, San Diego, CA, USA and TAK1 inhibitor 5z-7-oxozeanol from Bio-Techne Corporation, Minneapolis, MN, USA.
+ Open protocol
+ Expand
2

Comprehensive Cell Death Pathway Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols
Human recombinant TNF, BV6, GNE684, and GSK583 were all synthesized at Genentech. Emricasan was purchased from Selleck Chemicals (S7775), zVAD from ABclonal and LPS from InvivoGen (tlrl-3peLPS). The primary antibodies used were directed against: RIP1 (610459, BD Biosciences), pRIP1 S166 (#31122, Cell Signaling Technology (CST)), RIP3 (#15828, CST), pRIP3 (#91702, CST), MLKL (#MABc604, Millipore), pMLKL (#37333, CST), FADD (#05-486, Millipore), RIP2 (#22763, Santa Cruz), XIAP (M044-3, MBL, 66800-1-Ig, Proteintech), c-IAP2 (Genentech), c-IAP1/2 (#3400, R&D), caspase-1(Genentech), caspase-3 (#9661, 9662, CST), caspase-7(#9491, CST), caspase-8 (#8592, 9429, 4927, CST), caspase-11(#14340, CST), GSDMD (#50928, CST), IL-1b (AF-401-NA, R&D), JNK(#9252, CST), pJNK (#4668, CST), p65(#8242, CST), p-p65(#3033, CST), IkBa (#9242, CST), p-IkBa (#2859, CST), p38 (#9212, CST), p-p38 (#9211, CST), ERK (#4695, CST), p-ERK (#4370, CST), actin (A3853, Sigma), GapDH (#2118, CST), HSP90 (#4877, CST).
+ Open protocol
+ Expand
3

Immune Profiling of Lepirudin-treated PBMCs

Check if the same lab product or an alternative is used in the 5 most similar protocols
PBMC were isolated from whole-blood anticoagulated with lepirudin (Refludan®, Celgene). Recombinant C5a, ultrapure cholesterol, 1-propanol were purchased from Sigma-Aldrich, and TNF from Genentech. The following reagents were used for qRT-PCR analyses: RNeasy Mini kit (Qiagen), DNase and High Capacity RNA-to-cDNA Kit (Applied Biosystems) and PerfeCTa® qPCR FastMix™ (Quanta Biosciences). Assays were from Applied Biosystems: GAPDH (Hs99999905_m1), NLRP3 (Hs00918082_m1), TNF (Hs00174128_m1), and IL-1β (Hs01555410_m1) and primers from SYBR as shown in Supplementary Table I. Antibodies used for immunohistochemistry: polyclonal rabbit anti-human C1q (A0136, Dako), normal rabbit IgG (Ab105-C, R&D systems), goat anti-mouse conjugated to Alexa 647 (Invitrogen), anti-C5b-9 (DIA-011, Antibodyshop), anti-CD68 (Dako Agilent M0814), anti-CD20 (Dako Agilent M0755), anti-CD3 (Dako Agilent A0452), anti-CD15 (BD Pharmingen 559045), control IgG2a (BD Pharmigen). Antibodies used for flow cytometry included anti-CD14 FITC (M5E2), anti-CD4 PerCP-CY 5.5 (RPA-T4), anti-CD8 BV605 (SK1) and anti-CD19 BV711 (SJ25C1) and were purchased from BD Pharmigen, and anti-CD3 APC (UCHT1) were from Invitrogen. Fluorescence-minus-one (FMO) was used as control.
+ Open protocol
+ Expand

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

Sign up now

Revolutionizing how scientists
search and build protocols!