Journal: F1000Research

Article Title: The identification of high-performing antibodies for TDP-43 for use in Western Blot, immunoprecipitation and immunofluorescence

doi: 10.12688/f1000research.131852.2

Figure Lengend Snippet: Lysates of HAP1 (WT and TARDBP KO) were prepared and 50 μg of protein were processed for Western blot with the indicated TDP-43 antibodies. The Ponceau stained transfers of each blot are shown. Antibody dilutions were chosen according to the recommendations of the antibody supplier. Exceptions were given for antibody 80001-1-RR**, which was titrated to 1/1000 as the signal was too weak when following the supplier’s recommendations. When the concentration was not indicated by the supplier, which was the case for antibody 80002-1-RR**, we tested the antibody at 1/1000. Antibody dilution used: 10782-2-AP at 1/5000, 12892-1-AP at 1/1000, 800001-1-RR** at 1/1000, 80002-1-RR** at 1/1000, MAB7778* at 1/500, NBP1-92695* at 1/1000, 711051** at 1/1000, MA5-27828* at 1/1000, MA5-32627** at 1/1000, A19123** at 1/1000, 89789** at 1/1000, 89718** at 1/1000, GTX630196* at 1/500, GTX630197* at 1/500, ab109535** at 1/2000, ab133547** at 1/1000, ab190963** at 1/1000, ab254166** at 1/1000. Predicted band size: 45 kDa. *Monoclonal antibody, **Recombinant antibody.

Article Snippet: Bio-Techne , NBP1-92695 , 122117 , AB_11005586 , monoclonal , 3H8 , mouse , 1.00 , Wb, IF.

Techniques: Western Blot, Staining, Concentration Assay, Recombinant

Journal: F1000Research

Article Title: The identification of high-performing antibodies for TDP-43 for use in Western Blot, immunoprecipitation and immunofluorescence

doi: 10.12688/f1000research.131852.2

Figure Lengend Snippet: HAP1 WT and TARDBP KO cells were labelled with a green or a far-red fluorescent dye, respectively. WT and KO cells were mixed and plated to a 1:1 ratio in a 96-well plate with a glass bottom. Cells were stained with the indicated TDP-43 antibodies and with the corresponding Alexa-fluor 555 coupled secondary antibody including DAPI. Acquisition of the blue (nucleus-DAPI), green (identification of WT cells), red (antibody staining) and far-red (identification of KO cells) channels was performed. Representative images of the merged blue and red (grayscale) channels are shown. WT and KO cells are outlined on both channels with green and magenta dashed lines, respectively. Antibody dilutions were chosen according to the recommendations of the antibody supplier. Exceptions were given for antibodies 12892-1-AP, MA5-32627**, A19123**, 89789**, 89718** and ab133547** which were titrated to 1/400, 1/1000, 1/900, 1/30, 1/10 and 1/700, respectively, as the signals were too weak when following the supplier’s recommendations When the concentration was not indicated by the supplier, which was the case for antibodies 80001-1-RR**, GTX630196* and ab254166**, we tested antibodies at 1/700, 1/1000 and 1/500, respectively. At these concentrations, the signal from each antibody was in the range of detection of the microscope used. Antibody dilution used: 10782-2-AP at 1/400, 12892-1-AP at 1/250, 800001-1-RR** at 1/700, 80002-1-RR** at 1/250, MAB7778* at 1/500, NBP1-92695* at 1/1000, 711051** at 1/500, MA5-27828* at 1/1000, MA5-32627** at 1/1000, A19123** at 1/900, 89789** at 1/30, 89718** at 1/10, GTX630196* at 1/1000, GTX630197* at 1/1000, ab109535** at 1/30, ab133547** at 1/700, ab190963** at 1/800, ab254166** at 1/500. Bars = 10 μm. *Monoclonal antibody, **Recombinant antibody.

Article Snippet: Bio-Techne , NBP1-92695 , 122117 , AB_11005586 , monoclonal , 3H8 , mouse , 1.00 , Wb, IF.

Techniques: Staining, Concentration Assay, Microscopy, Recombinant

Journal: F1000Research

Article Title: The identification of high-performing antibodies for TDP-43 for use in Western Blot, immunoprecipitation and immunofluorescence

doi: 10.12688/f1000research.131852.2

Figure Lengend Snippet: Summary of the TDP-43 antibodies tested.

Article Snippet: Bio-Techne , NBP1-92695 , 122117 , AB_11005586 , monoclonal , 3H8 , mouse , 1.00 , Wb, IF.

Techniques: Concentration Assay, Recombinant

Journal: Data in Brief

Article Title: Genome sequence data of Burkholderia sp. IMCC1007 isolated from maize rhizosphere: A potential strain in fusaric acid mycotoxin biodegradation

doi: 10.1016/j.dib.2023.109204

Figure Lengend Snippet: Responsible genes found in the strain IMCC1007 genome.

Article Snippet: Gra , Granaticin polyketide synthase , Streptomyces violaceoruber ATCC 14980 T , 50.4.

Techniques:

Journal: Brain Sciences

Article Title: Identification and Validation of Novel Potential Pathogenesis and Biomarkers to Predict the Neurological Outcome after Cardiac Arrest

doi: 10.3390/brainsci12070928

Figure Lengend Snippet: Primer sequences.

Article Snippet: Anti-RPL27 , Proteintech , 14980-1-AP , 1:500.

Techniques: Sequencing

Journal: Brain Sciences

Article Title: Identification and Validation of Novel Potential Pathogenesis and Biomarkers to Predict the Neurological Outcome after Cardiac Arrest

doi: 10.3390/brainsci12070928

Figure Lengend Snippet: The information of the primary antibodies used in this study.

Article Snippet: Anti-RPL27 , Proteintech , 14980-1-AP , 1:500.

Techniques:

Journal: Journal of Orthopaedic Translation

Article Title: Comparative proteomic analysis identifies differentially expressed proteins and reveals potential mechanisms of traumatic heterotopic ossification progression

doi: 10.1016/j.jot.2022.04.003

Figure Lengend Snippet: 1 Selected hub Proteins in magenta module as Identified by Proteomics Analysis 2 Selected hub Proteins in turquoise module as Identified by Proteomics Analysis.

Article Snippet: Subsequently, the membranes were blocked in 5% nonfat milk or bovine serum albumin (BSA) for 1 ​h and subsequently probed overnight at the temperature of 4 ​°C with diluted primary antibodies against β-Actin (1:5000, T0022, Affinity biosciences, ChangZhou, China), UBA1(1:2000, ab180125, Abcam Inc, Cambridge, UK), EIF3E (1:1000, ab134958, Abcam Inc, Cambridge, UK), RPL27 (1:500, 14980-1-AP, Proteintech, Chicago, USA), RPL17 (1:5000, 67223-1-Ig, Proteintech, Chicago, USA), RPS28(1:500, 14796-1-AP, Proteintech, Chicago, USA).

Techniques:

Journal: Journal of Orthopaedic Translation

Article Title: Comparative proteomic analysis identifies differentially expressed proteins and reveals potential mechanisms of traumatic heterotopic ossification progression

doi: 10.1016/j.jot.2022.04.003

Figure Lengend Snippet: qPCR and immunoblot assay compared the differential expression of the UBA1, EIF3E, RPL27, RPS28, RPL17 in tissue samples of rat models and clinical specimens. The relative gene expression of UBA1, EIF3E, RPL27, RPS28, RPL17 was evaluated in (a) clinical specimens and (b) tissue samples of AT, PAT and control rat models using qRT-PCR (c, d) Western blots for UBA1, EIF3E, RPL27, RPS28, RPL17 on whole-tissue lysate from clinical specimens and control, AT, PAT rat models. ​β-Actin served as a loading control. The relative expression level of UBA1, EIF3E, RPL27, RPS28, RPL17 in (e) clinical specimens and (f) tissue samples of control, AT, PAT rat models quantified using ImageJ and normalized to b-actin is shown. All tests in triplicate, ∗ represent significant change in the expression level compared to control group, ∗P ​< ​0.05, ∗∗P ​< ​0.01.

Article Snippet: Subsequently, the membranes were blocked in 5% nonfat milk or bovine serum albumin (BSA) for 1 ​h and subsequently probed overnight at the temperature of 4 ​°C with diluted primary antibodies against β-Actin (1:5000, T0022, Affinity biosciences, ChangZhou, China), UBA1(1:2000, ab180125, Abcam Inc, Cambridge, UK), EIF3E (1:1000, ab134958, Abcam Inc, Cambridge, UK), RPL27 (1:500, 14980-1-AP, Proteintech, Chicago, USA), RPL17 (1:5000, 67223-1-Ig, Proteintech, Chicago, USA), RPS28(1:500, 14796-1-AP, Proteintech, Chicago, USA).

Techniques: Western Blot, Expressing, Quantitative RT-PCR

Journal: Journal of Orthopaedic Translation

Article Title: Comparative proteomic analysis identifies differentially expressed proteins and reveals potential mechanisms of traumatic heterotopic ossification progression

doi: 10.1016/j.jot.2022.04.003

Figure Lengend Snippet: Representative IHC staining of (a–e) UBA1, EIF3E, RPL27, RPS28, RPL17 in tendon tissue of control group and injury tissue from AT and PAT groups 2days, 7days, 14days, 28days, 10weeks after modeling. Original magnification is 20x. Inserts are approximately 4x magnified images of the boxed area. Scale bars: 250 ​μm.

Article Snippet: Subsequently, the membranes were blocked in 5% nonfat milk or bovine serum albumin (BSA) for 1 ​h and subsequently probed overnight at the temperature of 4 ​°C with diluted primary antibodies against β-Actin (1:5000, T0022, Affinity biosciences, ChangZhou, China), UBA1(1:2000, ab180125, Abcam Inc, Cambridge, UK), EIF3E (1:1000, ab134958, Abcam Inc, Cambridge, UK), RPL27 (1:500, 14980-1-AP, Proteintech, Chicago, USA), RPL17 (1:5000, 67223-1-Ig, Proteintech, Chicago, USA), RPS28(1:500, 14796-1-AP, Proteintech, Chicago, USA).

Techniques: Immunohistochemistry

Journal: Journal of Orthopaedic Translation

Article Title: Comparative proteomic analysis identifies differentially expressed proteins and reveals potential mechanisms of traumatic heterotopic ossification progression

doi: 10.1016/j.jot.2022.04.003

Figure Lengend Snippet: Representative IHC staining of (a–e) UBA1, EIF3E, RPL27, RPS28, RPL17 in ligament tissue of clinically relevant HO patients, clinically irrelevant HO patients and HO negative patients. Semiquantitative analysis of immunohistochemical staining of UBA1, EIF3E, RPL27, RPS28, RPL17 expression in tissue samples of control, AT and PAT rat models (f) and clinical specimens (g). ImageJ software was applied to calculate the average optical density. Original magnification is 20x. Inserts are approximately 4x magnified images of the boxed area. Scale bars: 250 ​μm n ​= ​3/group. ∗ represent significant change in the expression level compared to control group, ∗P ​< ​0.05, ∗∗P ​< ​0.01.

Article Snippet: Subsequently, the membranes were blocked in 5% nonfat milk or bovine serum albumin (BSA) for 1 ​h and subsequently probed overnight at the temperature of 4 ​°C with diluted primary antibodies against β-Actin (1:5000, T0022, Affinity biosciences, ChangZhou, China), UBA1(1:2000, ab180125, Abcam Inc, Cambridge, UK), EIF3E (1:1000, ab134958, Abcam Inc, Cambridge, UK), RPL27 (1:500, 14980-1-AP, Proteintech, Chicago, USA), RPL17 (1:5000, 67223-1-Ig, Proteintech, Chicago, USA), RPS28(1:500, 14796-1-AP, Proteintech, Chicago, USA).

Techniques: Immunohistochemistry, Immunohistochemical staining, Staining, Expressing, Software