Log In Sign up
The largest database of trusted experimental protocols

Amg655

Manufactured by Amgen
Visit Supplier
Sourced in United States

AMG655 is a laboratory equipment product manufactured by Amgen. It is designed for use in scientific research and analysis. The core function of AMG655 is to provide a platform for performing various experimental procedures and analyses in a controlled laboratory environment.

Automatically generated - may contain errors

Lab products found in correlation

Antibody coupling kit, by Thermo Fisher Scientific (2 mentions) Dynabead, by Thermo Fisher Scientific (2 mentions) Dynabead antibody coupling kit, by Thermo Fisher Scientific (1 mentions) Trail r2, by Cell Signaling Technology (1 mentions) Dynamag 2 magnetic rack, by Thermo Fisher Scientific (1 mentions) Entinostat ms 275, by Selleck Chemicals (1 mentions) Fugene hd transfection reagent, by Promega (1 mentions) Annexin 5 fitc, by BD (1 mentions) Recombinant human trail, by R&D Systems (1 mentions) Recombinant human trail, by Thermo Fisher Scientific (1 mentions) B ap15, by Cayman Chemical (1 mentions)

5 protocols using amg655

1

Antibody Sourcing for TRAIL-Induced Apoptosis

Check if the same lab product or an alternative is used in the 5 most similar protocols
Antibodies were sourced as follows: AMG655 (conatumumab) was sourced from Amgen Inc (Thousand Oaks, CA, USA); FADD mouse monoclonal antibody was from BD transduction laboratories (catalog #556402); N‐terminal specific procaspase 8 antibody was purchased from Abcam (catalog #ab32125); C‐terminal specific procaspase‐8 antibody was purchased from Enzo® life sciences (catalog # ALX‐804‐242‐C100); FLIP antibody (NF6; catalog # AG‐20B‐0056‐C100) was purchased from Adipogen; TRAIL‐R2 was purchased from Cell signalling technology (catalog #3696); and procaspase‐10 was purchased from MBL international (catalog #M059‐3). Horseradish peroxidase‐conjugated secondary antibodies were purchased from cell signalling technology; LI‐COR mouse (IRDye® 800CW) and rabbit (IRDye® 680CW) secondary antibodies were purchased from LI‐COR. MS‐275 (entinostat) was obtained from Selleck Chemicals (Newmarket, UK); annexin V‐FITC was obtained from BD biosciences; and z‐Val‐Ala‐Asp(OME)‐FMK (zVAD‐FMK) was purchased from Sigma‐Aldrich (Gillingham, UK). Transfections were completed using FuGENE® HD transfection reagent (Promega, UK). The Dynabead® antibody coupling kit and a Dynamag™‐2 magnetic rack were obtained from Life Technologies (Paisley, UK). FADD peptides were generated by Almac Sciences (Edinburgh, UK).
Humphreys L.M., Fox J.P., Higgins C.A., Majkut J., Sessler T., McLaughlin K., McCann C., Roberts J.Z., Crawford N.T., McDade S.S., Scott C.J., Harrison T, & Longley D.B. (2020). A revised model of TRAIL‐R2 DISC assembly explains how FLIP(L) can inhibit or promote apoptosis. EMBO Reports, 21(3), e49254.
+ Open protocol
+ Expand
2

DR5-Mediated Death-Inducing Signaling Complex Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols
AMG655 (Conatumumab) was a kind gift from Amgen. This fully humanised antibody, which recognises the extracellular region of DR5, was conjugated to Dynabeads® using the antibody coupling kit from Invitrogen as per manufacturer’s instructions. For the DR5 DISC assay, 30μL of Dynabeads® coated with 5μg of AMG655 was added to 2×106 cells and incubated for 1hr. The cells were then lysed in DISC buffer (0.2% NP-40, 20 mM Tris-HCL (pH 7.4), 150 mM NaCl, 10% glycerol). The AMG655-coated Dynabeads® were collected and washed 5 times in DISC buffer prior to resuspension in Laemmli buffer and analysis by Western blotting. Supernatants and inputs were also collected and analysed by Western blotting.
Majkut J., Sgobba M., Holohan C., Crawford N., Logan A., Kerr E., Higgins C., Redmond K., Riley J., Stasik I., Fennell D., Van Schaeybroeck S., Haider S., Johnston P., Haigh D, & Longley D. (2014). Differential affinity of FLIP and procaspase 8 for FADD’s DED binding surfaces regulates DISC assembly. Nature communications, 5, 3350.
+ Open protocol
+ Expand
3

DR5-Mediated Death-Inducing Signaling Complex Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols
AMG655 (Conatumumab) was a kind gift from Amgen. This fully humanised antibody, which recognises the extracellular region of DR5, was conjugated to Dynabeads® using the antibody coupling kit from Invitrogen as per manufacturer’s instructions. For the DR5 DISC assay, 30μL of Dynabeads® coated with 5μg of AMG655 was added to 2×106 cells and incubated for 1hr. The cells were then lysed in DISC buffer (0.2% NP-40, 20 mM Tris-HCL (pH 7.4), 150 mM NaCl, 10% glycerol). The AMG655-coated Dynabeads® were collected and washed 5 times in DISC buffer prior to resuspension in Laemmli buffer and analysis by Western blotting. Supernatants and inputs were also collected and analysed by Western blotting.
Majkut J., Sgobba M., Holohan C., Crawford N., Logan A., Kerr E., Higgins C., Redmond K., Riley J., Stasik I., Fennell D., Van Schaeybroeck S., Haider S., Johnston P., Haigh D, & Longley D. (2014). Differential affinity of FLIP and procaspase 8 for FADD’s DED binding surfaces regulates DISC assembly. Nature communications, 5, 3350.
+ Open protocol
+ Expand
4

Evaluating TRAIL-induced Cell Viability

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
Two thousand cells per well were seeded into 96-well plates and cultured for 72 h in the presence of increasing concentrations of recombinant human TRAIL (R&D systems, Minneapolis, USA), AMG655 (Amgen, Thousand Oaks, CA, USA) or APG880 (Apogenix, Heidelberg, Germany) as indicated. Cell viability was determined by Cristal Violet staining (Sigma-Aldrich, St. Louis, MO, USA) as previously described [33 (link)]. IC50 were calculated using GraphPad Prism v9.4.0 software modelization. Molecular weights are reported in Table S2.
Brion R., Gantier M., Biteau K., Taurelle J., Brounais-Le Royer B., Verrecchia F., Rédini F, & Guiho R. (2022). TRAIL-Based Therapies Efficacy in Pediatric Bone Tumors Models Is Modulated by TRAIL Non-Apoptotic Pathway Activation via RIPK1 Recruitment. Cancers, 14(22), 5627.
+ Open protocol
+ Expand
5

Apoptosis Induction Pathway Dissection

Check if the same lab product or an alternative is used in the 5 most similar protocols
b-AP15 was purchased from Cayman Chemical Company (Ann Arbor, MI). CFZ was purchased from Selleck Chemicals (Houston, TX). BTZ was originally provided by Millennium Pharmaceuticals (Cambridge, MA). CQ was purchased from Sigma Chemical Co. (St. Louis, MO). Baf A was purchased from LC laboratories (Woburn, MA). Human recombinant TRAIL was purchased from PeproTech, Inc. (Rocky Hill, NJ). AMG655 was supplied by Amgen, Inc (Thousand Oaks, CA). Other antibodies were the same as described previously45 (link).
Oh Y.T., Deng L., Deng J, & Sun S.Y. (2017). The proteasome deubiquitinase inhibitor b-AP15 enhances DR5 activation-induced apoptosis through stabilizing DR5. Scientific Reports, 7, 8027.
+ 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!