human α 2 antiplasmin  (Innovative Research Inc)

Journal: Scientific Reports

Article Title: Structure functional insights into calcium binding during the activation of coagulation factor XIII A

doi: 10.1038/s41598-019-47815-z

Figure Lengend Snippet: Effect of rFXIII-A calcium binding site mutants on its specific activity and substrate specificity. Panels a, b and c represent comparative bar graphs for Specific activity of intracellular lysates (wild type and variant) based on Photometric assay, interpolated FXIII-A activity based on Pentylamine incorporation assay and α-2 antiplasmin incorporation assay, respectively. In the event a variant is significantly different than the wild type, it is marked with a star (*) sign on top of the corresponding bar. Statistical significance is set at p < 0.05. Experiments were performed in duplicates, thrice (N = 6). Panel d shows the putative fibrinogen, α-2 antiplasmin and BAPA binding site residues on zymogenic FXIII-A monomeric and activated monomeric FXIII-A* structures along with two intermediate transition state structures. The structure backbone is depicted in grey colored ribbon format. The putative binding site region backbones are colored cyan for fibrinogen, yellow for BAPA and magenta for α-2 antiplasmin. The three calcium binding sites are also depicted on all four structures as stick models colored red (Cab1), blue (Cab2) and green (Cab3) respectively.

Article Snippet: Cross-linking of the α 2 -antiplasmin into the fibrin by rFXIII was detected using goat anti-human α 2 -antiplasmin antibody with a horse-radish peroxidase conjugate (Enzyme Research Laboratories) and 1, 2-diaminobenzene o -phenylenediamine (OPD; Dako).

Techniques: Binding Assay, Activity Assay, Variant Assay

Journal: Journal of thrombosis and haemostasis : JTH

Article Title: Factor XIII cotreatment with hemostatic agents in hemophilia A increases fibrin α-chain crosslinking

doi: 10.1111/jth.13887

Figure Lengend Snippet: As compared with FVIII treatment alone, FXIII cotreatment accelerates and increases α2-antiplasmin crosslinking. (A) Representative western blots for α2-antiplasmin crosslinking in recalcified hemophilic plasma treated as in Fig. 1. (B) Quantification of crosslinked α2-antiplasmin normalized to total α2-antiplasmin at t = 0 min (mean ± standard error of the mean [SEM], n = 7). Symbols are: buffer, closed circles; + FXIII, half-filled boxes; + FVIII, open triangles; + FVIII + FXIII, closed inverted triangles. *P < 0.05 versus buffer-treated, FXIII-treated and FVIII-treated samples. Two-way ANOVA with the Holm–Sidak multiple comparisons test was used to compare time and treatment. (C) Maximum rate of α2-antiplasmin crosslinking (mean ± SEM, n = 7, arbitrary units [AU] min−1). Treatments were compared by the use of repeated measures ANOVA with the Holm–Sidak multiple comparisons test.

Article Snippet: Polyclonal sheep anti-human α 2 -antiplasmin antibody was from Affinity Biologicals (Ancaster, Ontario, Canada).

Techniques: Western Blot, Clinical Proteomics

Journal: Journal of Cellular and Molecular Medicine

Article Title: A specific plasminogen activator inhibitor‐1 antagonist derived from inactivated urokinase

doi: 10.1111/jcmm.12875

Figure Lengend Snippet: Specificity of PAI trap for PAI ‐1. The inhibition of serpins by PAI trap in comparison to PAI ‐1. The ratio of PAI trap inhibition of the serpin studied compared to PAI ‐1 is presented as a function of PAI trap concentration. ( A ) PAI ‐2; ( B ) PN ‐1; ( C ) anti‐thrombin‐ III ; ( D ) 2‐antiplasmin.

Article Snippet: Antithrombin‐III, α‐thrombin, human α 2 ‐antiplasmin and human plasmin were from Haematologic Technologies, Essex Junction, USA.

Techniques: Inhibition, Comparison, Concentration Assay

Journal: Frontiers in Cellular Neuroscience

Article Title: Tissue plasminogen activator inhibits NMDA-receptor-mediated increases in calcium levels in cultured hippocampal neurons

doi: 10.3389/fncel.2015.00404

Figure Lengend Snippet: tPA inhibition of NMDA-induced calcium influx is independent of plasmin and lipoprotein receptor-related protein 1 (LRP-1). (A) Hippocampal cultures were preincubated with tPA (40 μg/ml; for 5 min) or α 2 -antiplasmin (140 nM; for 15 min) and tPA (for 5 min). Baseline Fluo-4 fluorescence was monitored for 15 s prior to the addition of NMDA (5 μM), at time = 0. Fluo-4 fluorescence was monitored for a further 45 s. Raw fluorescence values were converted to ΔF/F 0 , where F 0 is the average fluorescence over the first 15 s of recording prior to addition of agonist (baseline) and ΔF is F max −F 0 . (B) The responses in A were quantitated by measuring the AUC and are presented relative to the AUC for 5 μM NMDA (100%). (C) Hippocampal cultures were preincubated with tPA (40 μg/ml) for 5 min or receptor-associated protein (RAP) (500 nM; for 15 min) and tPA (for 5 min). Baseline Fluo-4 fluorescence was monitored for 15 s prior to the addition of NMDA (5 μM), at time = 0. Fluo-4 fluorescence was monitored for a further 45 s. Raw fluorescence values were converted to ΔF/F 0 , where F 0 is the average fluorescence over the first 15 s of recording prior to addition of agonist (baseline) and ΔF is F max −F 0 . (D) The responses in C were quantitated by measuring the AUC and are presented relative to the AUC for 5 μM NMDA (100%). RFU, Relative Fluorescent Units; n.s, not significant; *** p < 0.001. Error bar, SEM.

Article Snippet: Human α 2 -antiplasmin was purchased from MyBioSource (San Diego, CA, USA).

Techniques: Inhibition, Fluorescence