Platelet Activation Assay Protocol

Soluble collagen was from Corning (Corning, NY, USA). Prostaglandin I₂ (PGI₂) was from Cayman Chemical (Ann Arbor, MI, USA). Crosslinked collagen-related peptide (CRP-XL) was from R. Farndale (CambCol Laboratories, Cambridge University, UK). CHRONO-LUME^® detection reagent was from Chrono-Log Corporation (Havertown, PA, USA; #395). Pam2CSK4 and Pam3CSK4 were from Invivogen (#tlrl-pm2s-1 and #tlrl-pms). Tumor necrosis factor-α (TNF-α, #210-TA-020) was from R&D Systems. FSL-1 was from Tocris (#6011). Anti-TLR2 (#maba2-htlr2), anti-TLR6 (magb-htlr6) and control IgG (mabg1-ctrlm) and IgA (maba2-ctrl) antibodies were from Invivogen. All other reagents were from Sigma-Aldrich or previously named sources (48 (link)).

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Parra-Izquierdo I., Lakshmanan H.H., Melrose A.R., Pang J., Zheng T.J., Jordan K.R., Reitsma S.E., McCarty O.J, & Aslan J.E. (2021). The Toll-Like Receptor 2 Ligand Pam2CSK4 Activates Platelet Nuclear Factor-κB and Bruton’s Tyrosine Kinase Signaling to Promote Platelet-Endothelial Cell Interactions. Frontiers in Immunology, 12, 729951.

Publication 2021

Soluble collagen Prostaglandin I2 (PGI2) Pam2CSK4 Pam3CSK4 Tumor necrosis factor-α (TNF-α,

Antibodies Cayman Chrono lume Collagen Collagen related peptide Pam2csk4 Prostaglandin i2 Tlr2 Tlr6 Tnf α

Corresponding Organization : Oregon Health & Science University

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Variable analysis

independent variables

Soluble collagen (from Corning)
Prostaglandin I2 (PGI2) (from Cayman Chemical)
Crosslinked collagen-related peptide (CRP-XL) (from R. Farndale)
Pam2CSK4 (from Invivogen)
Pam3CSK4 (from Invivogen)
Tumor necrosis factor-α (TNF-α) (from R&D Systems)
FSL-1 (from Tocris)
Anti-TLR2 antibody (from Invivogen)
Anti-TLR6 antibody (from Invivogen)
Control IgG antibody (from Invivogen)
Control IgA antibody (from Invivogen)

dependent variables

Not explicitly mentioned

control variables

CHRONO-LUME® detection reagent (from Chrono-Log Corporation)
All other reagents (from Sigma-Aldrich or previously named sources)

positive controls

Not specified

negative controls

Control IgG antibody
Control IgA antibody

Annotations

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