Journal: Computational and Structural Biotechnology Journal

Article Title: Systematic screening and dynamic profiling of germline regulatory pathways and spermatogonial surface markers in a bivalve mollusc

doi: 10.1016/j.csbj.2025.01.023

Figure Lengend Snippet: Preparation and validation of the antiserum. (A) A flowchart illustrating the process of preparing the antiserum against PyFGFR extramembrane domains. (B) The antiserum was validated by Western blotting. The anti-6 × His tag monoclonal antibody, the antiserum, and the preimmune serum were utilized for the detection of PyFGFR in bacterial lysates. Similarly, the antiserum and the preimmune serum were employed for the detection of PyFGFR in protein extracts derived from scallop testes. The estimated molecular weights of the protein bands are indicated on the left. M, marker; B, bacterial lysates; T, testis proteins. (C) An immunohistochemical analysis was performed using three different types of serum: antiserum, antigen-preadsorbed antiserum, and preimmune serum. The analysis was conducted on the testis of P. yessoensis . Sg, spermatogonium; Sc, spermatocyte; St, spermatid; Sz, spermatozoon. (D) An immunofluorescent analysis was conducted using the antiserum. Some testicular cells exhibited red fluorescence on their surface. The nucleus was labeled with DAPI, which produces a blue fluorescence.

Article Snippet: Thereafter, it was incubated with rabbit anti-6 × His tag monoclonal antibody (PTM Biolabs, 1:5000), antiserum (1:5000), or preimmune serum (1:5000) overnight at 4°C.

Techniques: Biomarker Discovery, Western Blot, Derivative Assay, Marker, Immunohistochemical staining, Fluorescence, Labeling

Journal: Human Molecular Genetics

Article Title: Disruption of the methyltransferase-like 23 gene METTL23 causes mild autosomal recessive intellectual disability

doi: 10.1093/hmg/ddu115

Figure Lengend Snippet: Western blot for METTL23 protein: Purified METTL23-His8-tag protein (expressed in E. coli ) was loaded in lane 1 (50 ng) and 2–4 (100 ng). METTL23 proteins were immunoprecipitated from lymphoblastoid cells from healthy control (lane 5) and METTL23-mutant patient PK31 II:1 (lane 6) using polyclonal anti-METTL23. Immunoreactive bands were visualized using ( A ) rabbit polyclonal anti-METTL23-His8-tag antiserum, ( B ) rabbit polyclonal anti-His-tag (C-terminal) antibody and ( C ) sequence-specific anti-METTL23 (C17orf95) antibody as primary antibodies. Arrow (METTL23-His8-tag protein, 22.6 kDa); arrowhead (immunoprecipitated METTL23 protein, 21.5 kDa).

Article Snippet: Membranes were blocked with 5% (w/v) non-fat milk in TBST (Tris-buffered saline containing Tween 20) (25°C, 2 h) and incubated with either (i) rabbit polyclonal anti-METTL23 antiserum (1:2000 in 5%, w/v, BSA), (ii) rabbit polyclonal anti-His-tag (C-terminal) antibody (1:1000 in 5%, w/v, BSA, Relia Tech, Germany) and (iii) rabbit sequence-specific anti-METTL23 (C17orf95) antibody (1:300 in 5%, w/v, BSA, Abgent, Germany, raised against a synthetic peptide; position 137–166 amino acids) (4°C, overnight).

Techniques: Western Blot, Purification, Immunoprecipitation, Control, Mutagenesis, Sequencing