anti 6xhis alpha donor beads  (Revvity)

Journal: Neuropsychopharmacology

Article Title: Decreased striatal adenosine A 2A -dopamine D 2 receptor heteromerization in schizophrenia

doi: 10.1038/s41386-020-00872-9

Figure Lengend Snippet: a Illustration of the specific AlphaLISA protein–protein interaction assay designed for A 2A R–D 2 R heteromer identification and quantification in native tissue; anti-guinea pig-coated acceptor beads (red sphere) were generated to capture a guinea pig anti-A 2A R antibodies bound to the receptors within the membrane extract; anti-rabbit coated donor beads (blue sphere) capture the immune complexes between the rabbit anti-D 2 R antibodies and the receptors within the membrane extract; A 2A R–D 2 R heteromerization brings donor beads into close proximity (<200 nm) to the acceptor beads. The excitation of the donor beads at 680 nm generates singlet oxygen ( 1 O 2 ) molecules triggering a chemical reaction within the acceptor beads, which results in a sharp peak of fluorescent emission at 615 nm (figure designed using image templates from Servier Medical Art https://smart.servier.com/image-set-download/ ). b The A 2A R–D 2 R interaction capacity in membranes from saline- ( n = 10) and PCP- ( n = 10) treated mouse striatum (left panel) or from postmortem control ( n = 10) and schizophrenic ( n = 10; SCZ) caudate (right panel) was determined by AlphaLISA method (see ‘Methods’); the specific AlphaLISA signal (i.e., ∆AlphaLISA) was calculated as described in the ‘Methods’ section and expressed as percentage (mean ± SEM) of either the saline-treated mice or control subjects. ** P < 0.01 and *** P < 0.001, Student’s t -test.

Article Snippet: After 1 h incubation (in darkness at 22 °C), donor beads (anti-rabbit IgG AlphaLISA Donor beads, AS105D, PerkinElmer) were diluted in AlphaLISA buffer and 5 μL of diluted beads added to each well (final concentration 10 μg/mL).

Techniques: Protein Protein Interaction Assay, Generated, Membrane, Saline, Control

Journal: Neuropsychopharmacology

Article Title: Decreased striatal adenosine A 2A -dopamine D 2 receptor heteromerization in schizophrenia

doi: 10.1038/s41386-020-00872-9

Figure Lengend Snippet: a PPI impairment in saline- ( n = 10) and PCP-treated animals ( n = 10) with or without chronic treatment with haloperidol (Halo, 0.1 mg/kg/day for 5 days) or clozapine (Clz, 10 mg/kg/day for 5 days); results are expressed as percentage (mean ± SEM) of inhibition of ASR at the 75 dB amplitude prepulse acoustic stimulus; * P < 0.05, one-way ANOVA followed by Dunnett’s post hoc test compared to saline-treated mice. b A 2A R–D 2 R heteromerization in striatal membranes from the same animals shown in a determined by AlphaLISA method (see ‘Methods’); the specific AlphaLISA signal (i.e., ∆AlphaLISA) was calculated as described in the ‘Methods’ section and expressed as percentage (mean ± SEM) of the saline-treated mice. * P < 0.05 one-way ANOVA with Dunnett’s post hoc test when compared to saline-treated mice. c Representative immunoblot showing the expression of A 2A R, D 2 R and DAT in striatal membranes from animals from the same groups shown in a ; striatal membranes from PCP-treated mice were analyzed by SDS-PAGE (50 μg of protein/lane) and immunoblotted using guinea pig anti-A 2A R, rabbit anti-D 2 R, goat anti-DAT and rabbit anti-α-actinin antibodies (see ‘Methods’). d Relative quantification of A 2A R, D 2 R and DAT expression; the immunoblot protein bands corresponding to A 2A R, D 2 R, DAT and α-actinin from the same animals shown in a were quantified by densitometric scanning; values were normalized by the respective amount α-actinin in each lane to correct for protein loading. Results are expressed as percentage (mean ± SEM) of the corresponding saline-treated animal; *** P < 0.001, two-way ANOVA with Sidak’s post hoc test.

Article Snippet: After 1 h incubation (in darkness at 22 °C), donor beads (anti-rabbit IgG AlphaLISA Donor beads, AS105D, PerkinElmer) were diluted in AlphaLISA buffer and 5 μL of diluted beads added to each well (final concentration 10 μg/mL).

Techniques: Saline, Inhibition, Western Blot, Expressing, SDS Page, Quantitative Proteomics

Journal: Pharmaceutical Research

Article Title: Analytical and Functional Similarity Assessment of ABP 710, a Biosimilar to Infliximab Reference Product

doi: 10.1007/s11095-020-02816-w

Figure Lengend Snippet: Similarity Testing Plan and the Analytical Methods for the Structural and Functional Characterization of the Proposed Biosimilar ABP 710 and Infliximab Reference Products

Article Snippet: The AlphaLISA ® assay donor beads were coated with a hydrogel that contains phthalocyanine, a photosensitizer and streptavidin, which binds to biotinylated human IgG1 (Amgen).

Techniques: Functional Assay, Protein Concentration, Sequencing, Glycoproteomics, Hydrophilic Interaction Liquid Chromatography, Enzyme-linked Immunosorbent Assay, Spectroscopy, Activity Assay, Inhibition, Binding Assay, Imaging, Cytometry, Amplified Luminescent Proximity Homogenous Assay

Journal: Pharmaceutical Research

Article Title: Analytical and Functional Similarity Assessment of ABP 710, a Biosimilar to Infliximab Reference Product

doi: 10.1007/s11095-020-02816-w

Figure Lengend Snippet: Summary of Biological Activity Attributes for ABP 710, Infliximab (US), and Infliximab (EU)

Article Snippet: The AlphaLISA ® assay donor beads were coated with a hydrogel that contains phthalocyanine, a photosensitizer and streptavidin, which binds to biotinylated human IgG1 (Amgen).

Techniques: Activity Assay, Inhibition, Binding Assay, Enzyme-linked Immunosorbent Assay, SPR Assay, Imaging, Cytometry, Amplified Luminescent Proximity Homogenous Assay