huprot v3.1-human proteome microarray  (Cambridge Protein Arrays)

Journal: Bioactive Materials

Article Title: An intra articular injectable Mitocelle recovers dysfunctional mitochondria in cellular organelle disorders

doi: 10.1016/j.bioactmat.2024.09.021

Figure Lengend Snippet: Mitocelle can track and bind stressed mitochondria. (A) Schematic diagram of the human protein microarray analysis. (B) Mitocelle-interacting proteins were analyzed using the HuProt™ 3.1 human protein chip. The signal-to-noise ratio (SNR) for each spot was calculated as the ratio of the foreground-to-background signal. In addition, the GST signal intensity (red) was used for SNR normalization (left). A high-power image of NOX4 binding (white circles) is shown in the right panel. (C) Chord diagram visualizing the relationship between proteins with SNR > 1.0 and a list of mitochondrial, Golgi, and ER proteins. The SNR was calculated using the formula SNR = 20 log 10 (Is In −1 ), where ‘Is’ indicates the signal and ‘In’ indicates the noise. (D) Mouse chondrocytes treated with and without H 2 O 2 were analyzed by real-time live imaging of mitochondria (green) and Mitocelle (red) using a Celldiscoverer7 and an LSM900 confocal microscope. Intensity profiles of linear regions of interest are shown in the right panel. (E) To confirm mitochondrial dysfunction, we performed JC-1 staining and quantified JC-1 aggregates (red) and JC-1 monomers (green) as average intensities expressed in arbitrary units. Data are presented as means ± SD ( n = 5) and were assessed using one-way ANOVA with Bonferroni's test. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001.

Article Snippet: Protein microarray data were generated using the HuProt Human Proteome Microarray v3.1 (CDI Laboratories, Mayaguez, Puerto Rico), which contains more than 21,000 unique and individually purified full-length human protein clones.

Techniques: Microarray, Binding Assay, Imaging, Microscopy, Staining

Journal: Bioactive Materials

Article Title: An intra articular injectable Mitocelle recovers dysfunctional mitochondria in cellular organelle disorders

doi: 10.1016/j.bioactmat.2024.09.021

Figure Lengend Snippet: Mitocelle interferes with the NOX4-p22phox interaction that contributes to ROS generation. (A) HuProt™ 3.1 Human Protein chip was used to analyze the effects of free Cy5 and Cy5-labeled Mitocelle (5 μg/mL) on Mitocelle-interacting proteins. The signal-to-noise ratio (SNR) of each point was calculated as the ratio of foreground to background signal. Additionally, a high-power image of p22phox binding (white circle) is shown in the right panel. (B) The SNR >1.0 proteins included 10 proteins known to be involved in ROS generation. (C) Schematic illustration of the principle of FRET and the acceptor photobleaching used for FRET measurements; CFP (donor), YFP (acceptor). (D) FRET detection by acceptor photobleaching. Chondrocytes were transfected for 24 h with vectors encoding YFP-NOX4 and CFP-p22phox, Mitocelles were applied, and transfection was continued for an additional 24 h. Fluorescence images were collected using YFP and CFP channels before and after photobleaching. To better show the changes in CFP fluorescence, pre- and post-bleaching CFP images are presented using pseudocolor. (E) FRET efficiency was measured after acceptor bleaching. Data are presented as means ± SD ( n = 10) and were assessed using (E) Mann-Whitney U test. ∗∗∗∗ P < 0.0001.

Article Snippet: Protein microarray data were generated using the HuProt Human Proteome Microarray v3.1 (CDI Laboratories, Mayaguez, Puerto Rico), which contains more than 21,000 unique and individually purified full-length human protein clones.

Techniques: Labeling, Binding Assay, Transfection, Fluorescence, MANN-WHITNEY

Journal: Journal of Cellular and Molecular Medicine

Article Title: Human CPPED1 belongs to calcineurin‐like metallophosphoesterase superfamily and dephosphorylates PI3K‐AKT pathway component PAK4

doi: 10.1111/jcmm.16607

Figure Lengend Snippet: CPPED1‐interacting proteins obtained by human proteome microarray. Above‐threshold interactions are listed in order of highest to lowest binding affinity with CPPED1

Article Snippet: The HuProt TM v3.1‐Human Proteome Microarray (Cambridge Protein Arrays Ltd.) was used to identify protein interactions on an immobilized array.

Techniques: Microarray, Binding Assay, Derivative Assay, RNA Binding Assay, Ubiquitin Proteomics, Transduction

Journal: Journal of Cellular and Molecular Medicine

Article Title: Human CPPED1 belongs to calcineurin‐like metallophosphoesterase superfamily and dephosphorylates PI3K‐AKT pathway component PAK4

doi: 10.1111/jcmm.16607

Figure Lengend Snippet: KEGG pathway analysis of CPPED1 binding partners identified by human proteome microarray. Significant terms ( P <.05) are shown

Article Snippet: The HuProt TM v3.1‐Human Proteome Microarray (Cambridge Protein Arrays Ltd.) was used to identify protein interactions on an immobilized array.

Techniques: Binding Assay, Microarray

Journal: EBioMedicine

Article Title: Longitudinal serum autoantibody repertoire profiling identifies surgery-associated biomarkers in lung adenocarcinoma

doi: 10.1016/j.ebiom.2020.102674

Figure Lengend Snippet: The schematic diagram and work flow. Serum samples were collected longitudinally from the same patient at different time points before and after surgery. Representative samples were probed on the HuProt proteome microarray to identify candidates of surgery-associated antigens. A focused protein microarray with 14 identical subarrays was constructed by including the candidates of surgery-associated antigens. All the serum samples collected from the same patient were incubated on the focused microarray, some of the surgery-associated antigens were confirmed. (For interpretation of the references to color in this figure, the reader is referred to the web version of this article.)

Article Snippet: The HuProt human proteome microarrays (V3.1) from the same batch were purchased from CDI Laboratories, USA.

Techniques: Microarray, Construct, Incubation

Journal: EBioMedicine

Article Title: Longitudinal serum autoantibody repertoire profiling identifies surgery-associated biomarkers in lung adenocarcinoma

doi: 10.1016/j.ebiom.2020.102674

Figure Lengend Snippet: The serum autoantibody repertoires of the lung adenocarcinoma patients. (a) Representative proteome microarray results. (b) Pearson correlation coefficient matrix of IgG (upper) and IgM (lower). Each square represents the correlation coefficient of auto-antibody repertoires of two sera. (c) The correlations of the overall IgG signal intensities among samples from the same patient. (d) The correlations of the overall IgG signal intensities among samples from different patients. (e) The amounts of the positive autoantibodies (IgG and IgM) and the shared portion (the ratio of shared to total) of the three longitudinal sera from P1. (f) The ratios of shared/total positive autoantibodies of 5 sera for IgG and IgM in three groups, which are pre-operative (PxP), 1 month after surgery (PxA1) and 3 months after surgery (PxA3), respectively (x is the number of the samples, i.e ., 1, 2, 3, 4 or 5). (For interpretation of the references to color in this figure, the reader is referred to the web version of this article.)

Article Snippet: The HuProt human proteome microarrays (V3.1) from the same batch were purchased from CDI Laboratories, USA.

Techniques: Microarray

Journal: EBioMedicine

Article Title: Longitudinal serum autoantibody repertoire profiling identifies surgery-associated biomarkers in lung adenocarcinoma

doi: 10.1016/j.ebiom.2020.102674

Figure Lengend Snippet: Summary of surgery-associated candidate autoantigens.

Article Snippet: The HuProt human proteome microarrays (V3.1) from the same batch were purchased from CDI Laboratories, USA.

Techniques: Immunopeptidomics, Microarray

Journal: EBioMedicine

Article Title: Longitudinal serum autoantibody repertoire profiling identifies surgery-associated biomarkers in lung adenocarcinoma

doi: 10.1016/j.ebiom.2020.102674

Figure Lengend Snippet: Several serum autoantibodies with level decreased in response to surgery were validated . (a) The serum levels of anti- LSP1 IgG in samples of P1 according to the results of HuProt microarray. The gray arrows indicate the spots of LSP1 in the microarray, and the corresponding quantitative signal intensities are shown in the chart on the right. (b) The microarray segments of anti-LSP1 IgG for all the 14 sera from P1 detected by the focused microarray, the autoantibody levels of two unrelated proteins, i.e., RHOD and HRAS, were included as controls. (c–h) Other examples, e.g ., Anti-LSP1 IgG, anti-RGS20 IgM, anti-SNRPA IgM, anti-SPP1 IgG, anti-PVALB IgM and Anti-CDH12 IgM. “Days” mean days after surgery. All experiments were performed at least twice, the mean and standard deviations were calculated.

Article Snippet: The HuProt human proteome microarrays (V3.1) from the same batch were purchased from CDI Laboratories, USA.

Techniques: Microarray