Anti GST  

ELISA results of purified Goat anti-GST Antibody Biotin Conjugated tested against purified GST. Each well was coated in duplicate with 1.0 μg of GST (p/n 000-001-200). The starting dilution of antibody was 5μg/ml and the X-axis represents the Log10 of a 3-fold dilution. This titration is a 4-parameter curve fit where the IC50 is defined as the titer of the antibody. Assay performed using 3% fish gelatin as blocking buffer, Streptavidin Peroxidase Conjugated and TMB substrate p/n TMBE-1000

(G？I) GMF and VCA interfere with the SPIN90‐Arp2/3 interaction in the absence of filament. (G) Immunoblots from pull‐down assays where GST beads, decorated with GST or GST‐SPIN90‐Cter (the functional domain of SPIN90), were incubated with 0.4 μΜ Arp2/3 and gradient concentrations of GMF or VCA for 1 h at room temperature. After the unbound protein was washed out, the amount of Arp2/3 attached to the beads was detected by anti‐ArpC2 antibody. Ponceau red straining of the membrane verified that the beads were loaded with equal amounts of GST or GST‐SPIN90 (Appendix Fig？S2). (H, I) Quantification of the amount of ArpC2 remaining on the beads in different conditions. Data information: In all graphs (H and I): the bars indicate the average values, and the error bars indicate the standard deviations, from three independent repeats of each experiment (data points). Fig 4. PMID:？36939020

PEG passivation hinders non-specific protein adsorption. (a) Typical TIRF image of YFP-tagged protein pulled down using polyclonal anti-GFP antibody from the lysate (a, left). Non-specific binding of the protein to the PEG passivated surface (a, second from right) is comparable to the blank (a, second from left). Significant non-specific adsorption of YFP is observed when the same amount of lysate is added to surface passivated with 1 mg ml−1？BSA (a, right). Scale bar is 5 μm. (b) Bar graph with average number of fluorophores per image. Error bars represent standard deviation of the mean across > 20 images. Fig 2. PMID:？22322217

TAP/tapasin ratio is 1:2 in human cells. C, representative fluorescent images (left panel) and quantitation (right panel) of tapasin-YFP pulldown in slides coated with goat anti-GFP, the anti-TAP1 antibody 148.3, no antibody, or a control antibody (mouse anti-FLAG). To achieve an optimal density of molecules for imaging, the .220.B4402.tapasin-YFP lysate was diluted 1000-fold (anti-GFP), 20-fold (anti-TAP1), or 10-fold (no antibody and anti-FLAG) in 1% digitonin lysis buffer. The？scale bar？on the fluorescent images represents 5 μm. The quantitative results are expressed as the means plus the S.D. of at least 20 imaging areas (2500 μm2？each) in one representative experiment.？ Fig 1. PMID:？22829594

Characterization of skeletal muscle pathology in？Tg(ACTA1？D286G-eGFP)？high？zebrafish.？a？Skeletal muscle expressing？i？mosaic ACTA1D286G-eGFP, and？ii？overlaid with a light microscopy image of the same section showing Gomori trichrome staining, and？iii？enlarged.？Dark regions？(indicative of nemaline bodies) of disrupted muscle correspond to eGFP expression (arrows).？b？Correlative light and electron microscopy of？Tg(ACTA1？D286G-eGFP)？high？fish muscle at 2 dpf.？b？i？Fluorescent image and corresponding？ii？electron microscopy image of skeletal muscle section containing a dense, elongated nemaline body (arrow) and enlarged in (iii). Sections are matched using nuclei positions (asterisk,？plus？and？hash).？c？i？Accumulations of actin filaments (arrowheads) and？ii？diffuse regions of filamentous actin (arrowheads), as well as？ii？disrupted sarcomeric regions are evident in？Tg(ACTA1？D286G-eGFP)？high？skeletal muscle, at 2 dpf unlike the？iii？uniform sarcomeres observed in？Tg(ACTA1-eGFP)？zebrafish. Fig 2. PMID:？25931053

Localizations of Lgr5+？stem cells and pMET in the corpus epithelium of the adult stomach. (A) Immunohistochemical staining for GFP and pMET. (B) Double immunofluorescence staining for GFP and pMET. In (A), black arrows, Lgr5-driven GFP; red arrowheads, pMET in the glandular base region; yellow arrowheads, pMET in the isthmus and/or surface regions. In (B), white arrows indicate the co-localization of pMET and GFP in Lgr5+？stem cells. The images in the lower panel are magnified images of the boxed areas in the upper panel. Tissues were obtained from Lgr5-DTR-EGFP mice. Scale bars represent 200 μm. (C) The numbers of GFP- and pMET-positive cells per corpus gland. These values were obtained by counting in 10 glands (n？= 10). Data were tested for significance using an unpaired two-tailed？t-test. NS, not significant. Fig 6. PMID:？31212972

Confocal images of AAV9-hSyn and AAV9AU-hSyn mediated transduction in the rat striatum.？(A？C)？AAV9-JetI-GFP gene expression primarily co-localizes with the oligodendrocyte marker, Olig2, while？(D？F)？the general lack of GFP co-localization is with the neuronal marker, NeuN. In contrast？(G？I),？AAV9AU-JetI-GFP gene expression exhibits substantial co-localization with NeuN, but little co-localization with Olig2. Scale bars equal 20？μm. AAV, adeno-associated virus; GFP, green fluorescent protein. Fig 1. PMID:？32940068