rabbit anti-synapsin ii polyclonal antibody 56382-100  (QED Bioscience)

Journal: Brain, Behavior, & Immunity - Health

Article Title: Synapsin autoantibodies during pregnancy are associated with fetal abnormalities

doi: 10.1016/j.bbih.2023.100678

Figure Lengend Snippet: High prevalence of synapsin-I autoantibodies in pregnant women. (A) Representative examples of immunofluorescence stainings with human sera at 1:300 dilution (green) with (top row) or without binding (bottom row) to HEK293 cells overexpressing human synapsin-Ib. Protein expression is confirmed with a commercial synapsin-I/II antibody (red). Nuclei are stained with DAPI (blue). Scale bar = 20 μm. (B) Frequencies of synapsin-Ib autoantibodies in pregnant women and control cohorts, as determined by CBA. CBA scores: 0 = no binding, 1 = unspecific signal, 2 = intensive binding (positive); vertical dotted line represents cut-off for positivity. (C) Representative immunoblots of wild type (wt) and Syn1/2/3 triple knock out (TKO) mice cortex homogenates with a commercial synapsin-I/II antibody as positive control and sera from CBA-positive pregnant women (1:200 dilution). The strong band at ∼90 kDa corresponding to the molecular weight of synapsin-Ia/Ib was detected in wild type but not in TKO mouse tissue. Another band at ∼50 kDa likely represents the synapsin-IIb isoform or breakdown products of synapsin-I. Detection of GAPDH served as loading control. (D) Immunofluorescence staining on rat hippocampal neurons demonstrated strong IgG binding of human sera (green) in a characteristic synaptic pattern (insert shows high magnification, MAP2 [blue] staining for better visualization of neuronal processes). The punctate staining completely overlapped with a commercial synapsin antibody (red, insert for high magnification).

Article Snippet: A rabbit polyclonal synapsin-I/II antibody (Synaptic Systems, #106002) served as positive control to detect synapsin isoforms in brain lysates and transfected HEK cells.

Techniques: Immunofluorescence, Binding Assay, Expressing, Staining, Western Blot, Knock-Out, Positive Control, Molecular Weight

Journal: Brain, Behavior, & Immunity - Health

Article Title: Synapsin autoantibodies during pregnancy are associated with fetal abnormalities

doi: 10.1016/j.bbih.2023.100678

Figure Lengend Snippet: Strong, titer-dependent association of synapsin-I autoantibodies with abnormalities of fetal development during pregnancy. (A-B) Several ultrasound parameters and status of having previous children with neuropsychiatric disorders were significantly associated with the presence of synapsin-I autoantibodies in serum of pregnant mothers (A), with odds ratios of 3–7 (B). Statistical analysis used Fisher's exact test when expected frequencies were <5 or Chi-square test with expected frequencies ≥5, * representing p < 0.05, **p < 0.01 and ***p < 0.001. (C) Synapsin autoantibodies reached the fetal circulation and were present with similar titers (±1 titer step, in one case 2 titer steps) in umbilical cord blood. (D) Synapsin autoantibodies were of IgG1 and/or IgG3 subclass in almost all pregnant women. (E) While the autoantibody titer alone did not discriminate between pathological and normal findings during fetal development (left), the combination of IgG1 subclass with high-level titers (≥1:30,000) was significantly associated with abnormal measurements (right).

Article Snippet: A rabbit polyclonal synapsin-I/II antibody (Synaptic Systems, #106002) served as positive control to detect synapsin isoforms in brain lysates and transfected HEK cells.

Techniques:

Journal: Brain, Behavior, & Immunity - Health

Article Title: Synapsin autoantibodies during pregnancy are associated with fetal abnormalities

doi: 10.1016/j.bbih.2023.100678

Figure Lengend Snippet: Detection of synapsin-I autoantibodies by ELISA, immunoblotting and microarray-based epitope mapping. (A) ELISA-based screening of all human sera using recombinant synapsin-I correlated with autoantibody binding in the CBA. (B) Immunoblots of synapsin-Ib-transfected HEK293 cells confirm that some human sera (1:200 dilution) strongly detect the synapsin-Ib band (middle), while in others binding to non-conformational epitopes is lost (right). Commercial synapsin-I/II antibody served as positive control (left, GAPDH is exemplarily shown; UT = untransfected cells). (C) An overlapping peptide library containing 20-mer peptides with an off-set of three amino acids was designed to fully cover synapsin-I primary sequence. Microarrays were probed with autoantibody-positive and -negative serum. The representative example shows detection of IgG epitope 2 within the synapsin-I slides and a representative negative control serum. Heat map visualization: IgG binding intensities are displayed in grayscale (0 = no binding, 1 = maximal binding). The core shared motif is highlighted in red. (D) Synapsin-I domain architecture together with a cartoon representation of the alpha-fold model of synapsin-I visualized in Pymol. Mapped synapsin-I autoantibody epitopes (1–5, marked in red) are located in domain A-B (epitope 1, peptide sequence: 28 PQPPPPPPGAH 38 ), domain C (epitope 2, 124 TDWAKYFKGKK 134 ) and domain D (epitope 3: 472 PGPQRQGPPLQ 482 ; 4: 583 GGQQRQGPPQK 592 ; 5: 610 VPRTGPPTTQQPRP 623 ).

Article Snippet: A rabbit polyclonal synapsin-I/II antibody (Synaptic Systems, #106002) served as positive control to detect synapsin isoforms in brain lysates and transfected HEK cells.

Techniques: Enzyme-linked Immunosorbent Assay, Western Blot, Microarray, Recombinant, Binding Assay, Transfection, Positive Control, Sequencing, Negative Control