tris glycine sds page gel  (Bio-Rad)

Journal: bioRxiv

Article Title: Component A2 is a redox-sensitive archaeal ATPase activated by methyl-coenzyme M reductase

doi: 10.64898/2026.03.18.712670

Figure Lengend Snippet: (a) Genomic arrangement of the MCR activation operon in M. acetivorans with scale bar depicting 1 kilobase pair. (b) Structure of the putative MCR activation complex (PDB: 9H1L) with MCR in orange, component A2 (with relevant domains highlighted) in blue, and the other methanogenesis marker proteins in gray. NBD refers to N ucleotide B inding D omain and ZBM refers to Z inc B inding M otif. (c) Log 2 transformed FPKM ( F ragments P er K ilobase of transcript per M illion mapped reads) of the MCR activation operon and the MCR operon in M. acetivorans grown in high-salt (HS) minimal medium supplemented with trimethylamine (TMA) at 37 °C. (d) Genotype of an M. acetivorans strain expressing a second copy of component A2 in trans under the control of a tetracycline inducible promotor. (e) Anti-FLAG immunoblot showing the inducible production of component A2 upon the addition of 100 µg/mL tetracycline (tet) to the growth medium in crude and soluble cell lysates with 13.9 µg total protein loaded into each lane. (f) Anti-FLAG immunoblot of aerobic affinity-purification of A2 with a Streptactin resin. The lanes represent the following: (1) ladder, (2) cell lysate, (3) flow-through, (4) first wash, (5) second wash, (6) first elution, (7) second elution, and (8) third elution. (g) SDS-PAGE gel showing anaerobic purification of full-length TAP-tagged component A2 (64 kDa with tag). (h) Anaerobic ATPase assay of component A2 alone (blue), MCR alone (purple), and component A2 combined with MCR (orange). Each reaction contained 500 µg/mL of each protein indicated with 200 µM ATP, 10 mM MgCl 2 , 20 mM HEPES, 300 mM NaCl, and 1% glycerol. Reactions were incubated at 37 °C. Inorganic phosphate production was measured at 0, 15, 30, and 60 minutes using the malachite green reagent. (I) Aerobic ATPase assay of component A2 alone (blue), MCR alone (purple), and component A2 combined with MCR (orange). Proteins were purified anaerobically then removed from the anaerobic chamber and reactions were set up on the bench top. The same assay conditions as (c) were used, but time points were taken at 0, 30, and 60 minutes. Error bars represent the standard deviation of three technical replicates for each reaction.

Article Snippet: Samples were then loaded into 12% precast Tris-Glycine denaturing SDS-PAGE gels (Mini-PROTEAN TGX, Bio-Rad, Hercules, CA, USA).

Techniques: Activation Assay, Marker, Transformation Assay, Expressing, Control, Western Blot, Affinity Purification, SDS Page, Purification, ATPase Assay, Incubation, Standard Deviation

Journal: bioRxiv

Article Title: Inactivation of the RB1 and PTPN14 tumor suppressors cooperatively enables the carcinogenic activity of the human papillomavirus E7 oncoprotein

doi: 10.64898/2026.03.16.712171

Figure Lengend Snippet: Recombinant MHT-E7 proteins were purified from bacteria. (A) Size exclusion chromatogram (SEC) profiles of the different E7 proteins used for affinity measurements. Fractions between the dotted lines were pooled and used for downstream BLI assays. (B) SDS-PAGE of pooled fractions for each MHT-E7.

Article Snippet: Equal amounts of protein were resolved using Mini-Protean or Criterion Tris/Glycine SDS-PAGE gels (Bio-Rad) and transferred to polyvinylidene difluoride (PVDF) membrane.

Techniques: Recombinant, Purification, Bacteria, SDS Page

Journal: bioRxiv

Article Title: Helicobacter pylori allelic variation in cell surface genes influences human exoproteome binding and stomach tissue adherence

doi: 10.64898/2026.03.06.710112

Figure Lengend Snippet: (A-E) SDS-PAGE of purified LPS stained with ProQ Emerald 300 kit. Higher mobility LPS denoted ($). (A) Previously described H. pylori mouse-adapted strains and their parent human isolate. (B-E) PMSS1, SS1, and mouse outputs of PMSS1 at indicated times post-infection in C57BL6/N (black) or GIM mice (red). (F) Predicted amino acid variants of the α-( , )-fucosyltransferase FutB, the putative galactosyltransferase HPYLPMSS1_0826, G27 FutC (HpG27_0086), and the HpG27_579/580 loci from the subset of strains we sequenced. Strains and isolates are indicated next to the allele they are predicted to encode based on WGS.

Article Snippet: Samples were then run on 4-15% gradient SDS-PAGE Tris-glycine gels (Bio Rad Mini-PROTEAN TGX) with discontinuous running buffer (cathode: 0.1 M Tris, 0.1 M glycine, 0.1% SDS, pH 8.25; anode: 0.2 M Tris-HCl, pH 8.9) at 100 V until the dye front was near the bottom.

Techniques: SDS Page, Purification, Staining, Infection

Journal: bioRxiv

Article Title: Helicobacter pylori allelic variation in cell surface genes influences human exoproteome binding and stomach tissue adherence

doi: 10.64898/2026.03.06.710112

Figure Lengend Snippet: (A) SDS-PAGE of purified LPS from a panel of deletions subset of the characterized G27 glycosyltransferases. Loss of O-antigen production was identified in wecA, wzk, HpG27_1046, HpG27_0437, HpG27_0094, and HpG27_1235 deletions. (B) Schematic of the G27 LPS structure with enzymes responsible for the addition of each unit, adapted from Li et al. . (C) Adherence of fluorescein-labeled H. pylori to FFPE GIM mouse stomach tissue. Number of adhered bacteria per area of tissue plotted in GraphPad Prism. Purple - values greater than one standard deviation above wild type average (dotted line), green - values one standard deviation below. Sequential tissue slices from the same animal graphed with the same symbol.

Article Snippet: Samples were then run on 4-15% gradient SDS-PAGE Tris-glycine gels (Bio Rad Mini-PROTEAN TGX) with discontinuous running buffer (cathode: 0.1 M Tris, 0.1 M glycine, 0.1% SDS, pH 8.25; anode: 0.2 M Tris-HCl, pH 8.9) at 100 V until the dye front was near the bottom.

Techniques: SDS Page, Purification, Labeling, Bacteria, Standard Deviation