Journal: Scientific Reports

Article Title: Tracing the invisible mutant ADNP protein in Helsmoortel-Van der Aa syndrome patients

doi: 10.1038/s41598-024-65608-x

Figure Lengend Snippet: Overview of the reported ADNP antibodies with indication of the observed molecular weights and tested sample materials.

Article Snippet: Adult human brain (NB820-59177), human brain cerebellum (NB820-59180), human brain frontal lobe (NB820- 59186), and human brain hippocampus whole tissue lysates (NB820-59187) from a healthy 45-year-old male subject were purchased from Novus Biologicals.

Techniques: Molecular Weight, Transfection, Expressing, Construct

Journal: Scientific Reports

Article Title: Tracing the invisible mutant ADNP protein in Helsmoortel-Van der Aa syndrome patients

doi: 10.1038/s41598-024-65608-x

Figure Lengend Snippet: A polyclonal N-terminal ADNP antibody from Aviva Systems detects ADNP specifically in murine and rat tissues and suggests proteolytic processing of the protein in the human brain. Cerebellum, frontal cortex or lobe, hippocampus and whole brains of control mice, rats and humans were lysed in RIPA buffer and used as protein samples for the assessment of N-terminal antibody of Aviva systems. (A – C) The predicted molecular weight of ADNP is 124 kDa. The antibody recognizes ADNP in a range of 145 kDa with (E) additional lower mass signal of 85 kDa in all human brain regions. (B – D – F) Western blot analysis of the blocking peptide competition assay. Supplementation of the immunization peptide in a 5 × excess to antibody concentration reduced the signal observed at 145 kDa in all tested cell lines. Importantly, the 85 kDa band suggestive for proteolytic cleavage as well as degraded ADNP signal disappeared completely after immunization peptide supplementation. GAPDH was used as a loading control.

Article Snippet: Adult human brain (NB820-59177), human brain cerebellum (NB820-59180), human brain frontal lobe (NB820- 59186), and human brain hippocampus whole tissue lysates (NB820-59187) from a healthy 45-year-old male subject were purchased from Novus Biologicals.

Techniques: Control, Molecular Weight, Western Blot, Blocking Assay, Competitive Binding Assay, Concentration Assay

Journal: Scientific Reports

Article Title: Tracing the invisible mutant ADNP protein in Helsmoortel-Van der Aa syndrome patients

doi: 10.1038/s41598-024-65608-x

Figure Lengend Snippet: Different C-terminal ADNP antibodies detect ADNP in the range of 150 kDa and suggest proteolytic processing of the protein in the brain. Cerebellum, frontal cortex or lobe, hippocampus and whole brains of control mice, rats, and humans were lysed in RIPA buffer and used as protein samples for the assessment with three C-terminal antibodies with the optimized dilutions listed in Table . GAPDH was used as a loading control. The predicted molecular weight of ADNP is 124 kDa. ( A ) C ) Murine samples indicate detection of ADNP in the range of 150 kDa with bands suggesting proteolytic processing at 50 kDa. ( D – F ) Rat samples indicate detection of ADNP in the range of 150 kDa with bands indicating proteolytic processing at 82 kDa after incubation with the C-terminal Abcam antibody. ( G – I ) Human brain samples indicate detection of ADNP at different molecular weights of 124 – 150 kDa in the adult frontal lobe and hippocampus and highlight the antibody differences in detection of ADNP. The three tested antibodies showed strong band signals at lower molecular weights, which could indicate proteolytic cleavage or degradation of the protein.

Article Snippet: Adult human brain (NB820-59177), human brain cerebellum (NB820-59180), human brain frontal lobe (NB820- 59186), and human brain hippocampus whole tissue lysates (NB820-59187) from a healthy 45-year-old male subject were purchased from Novus Biologicals.

Techniques: Control, Molecular Weight, Incubation

Journal: Molecular cancer therapeutics

Article Title: Potency-matched dual cytokine-antibody fusion proteins for cancer therapy

doi: 10.1158/1535-7163.MCT-17-0211

Figure Lengend Snippet: A microarray containing normal tissue specimens (left) and their tumoral counterpart (right) was stained with a biotinylated preparation of the fully human IL2-F8-TNFmut (green, Alexa Fluor 488), cell nuclei were counterstained with DAPI (blue), 20x magnification, scale bars = 100µm.

Article Snippet: Frozen tumor and normal tissue specimens in microarray format were obtained from Amsbio and stained with a biotinylated preparation of the fully human IL2-F8-TNF mut fusion protein and detected with Streptavidin-AlexaFluor488 (Invitrogen {"type":"entrez-protein","attrs":{"text":"S11223","term_id":"112468","term_text":"pir||S11223"}} S11223 ).

Techniques: Microarray, Staining