Journal: Journal of Anatomy

Article Title: Scar-free cutaneous wound healing in the leopard gecko, Eublepharis macularius

doi: 10.1111/joa.12368

Figure Lengend Snippet: Summary table of the optimised immunohistochemistry and immunofluorescence protocols for proteins of interest

Article Snippet: Slides were then coverslipped using Cytoseal (TM) (Fischer Scientific). table ft1 table-wrap mode="anchored" t5 Table 1 caption a7 Antigen Type Retrieval Block Primary Secondary HRP DAB WE6 IHC 12 min citrate buffer 3% NGS 1.5 h RT Mouse anti-WE6 1 : 5 (Developmental Studies Hybridoma Bank WE6-s) Biotinylated goat anti-mouse 1 : 200 (Vector Laboratories BA-9200) 1 : 200 40 s PCNA IHC None 3% NGS 1 h RT Rabbit anti-PCNA 1 : 500 (Santa Cruz Biotechnology, Inc. sc-7907) Biotinylated goat anti-rabbit 1 : 200 (Jackson ImmunoResearch Laboratories, 111-066-003) 1 : 200 25 s VEGF IHC 12 min citrate buffer 3% NGS 1 h RT Rabbit anti-VEGF 1 : 100 (Santa Cruz Biotechnology, Inc. sc-152) Biotinylated goat anti-rabbit 1 : 500 (Jackson ImmunoResearch Laboratories, 111-066-003) 1 : 200 25 s TSP-1 IHC 12 min citrate buffer 3% NGS 1 h RT Mouse anti-Thrombospondin 1 1 : 50 (Santa Cruz Biotechnology, sc-59887) Biotinylated goat anti-mouse 1 : 500 (Vector Laboratories BA-9200) 1 : 200 40 s vWF IF None 3% NGS 1 h RT Rabbit anti-vonWillebrand Factor 1 : 500 (Dako Canada, A0082) Cy3 goat anti-rabbit 1 : 400 (Jackson ImmunoResearch Laboratories, 111-165-144) α-SMA IF None 3% NGS 1 h RT Mouse anti-α-Actin 1 : 400 (Santa Cruz Biotechnology, sc-32251) Goat anti-mouse AlexaFluor-488 1 : 400 (Life Technologies, A-11001) Open in a separate window Summary table of the optimised immunohistochemistry and immunofluorescence protocols for proteins of interest Immunofluorescence To visualise blood vessels, double-labelled immunofluorescence, for both vonWillebrand factor (vWF; expressed by endothelial cells) and α-smooth muscle actin (α-SMA; expressed by perivascular cells), was performed.

Techniques: Immunohistochemistry, Immunofluorescence, Blocking Assay, Plasmid Preparation, Immunohistochemistry-IF

Journal: Acta Crystallographica. Section F, Structural Biology Communications

Article Title: Neutron crystallographic study of heterotrimeric glutamine amidotransferase CAB

doi: 10.1107/S2053230X19000220

Figure Lengend Snippet: Macromolecule-production information (Nakamura et al. , 2006 ▸ , 2010 ▸ )

Article Snippet: Macromolecule-production information is given in Table 1 . table ft1 table-wrap mode="anchored" t5 Table 1 caption a7 Source organism S. aureus Mu50 Cloning site NcoI/XhoI Expression vector pET-28b(+) vector (Novagen) Expression host E. coli B834 strain Complete amino-acid sequence of the construct produced GatA MSIRYESVENLLTLIKDKKIKPSDVVKDIYDAIEETDPTIKSFLALDKENAIKKAQELDELQAKDQMDGKLFGIPMGIKDNIITNGLETTCASKMLEGFVPIYESTVMEKLHKENAVLIGKLNMDEFAMGGSTETSYFKKTVNPFDHKAVPGGSSGGSAAAVAAGLVPLSLGSDTGGSIRQPAAYCGVVGMKPTYGRVSRFGLVAFASSLDQIGPLTRNVKDNAIVLEAISGADVNDSTSAPVDDVDFTSEIGKDIKGLKVALPKEYLGEGVADDVKEAVQNAVETLKSLGAVVEEVSLPNTKFGIPSYYVIASSEASSNLSRFDGIRYGYHSKEAHSLEELYKMSRSEGFGKEVKRRIFLGTFALSSGYYDAYYKKSQKVRTLIKNDFDKVFENYDVVVGPTAPTTAFNLGEEIDDPLTMYANDLLTTPVNLAGLPGISVPCGQSNGRPIGLQFIGKPFDEKTLYRVAYQYETQYNLHDVYEKL GatB MHFETVIGLEVHVELKTDSKMFSPSPAHFGAEPNSNTNVIDLAYPGVLPVVNKRAVDWAMRAAMALNMEIATESKFDRKNYFYPDNPKAYQISQFDQPIGENGYIDIEVDGETKRIGITRLHMEEDAGKSTHKGEYSLVDLNRQGTPLIEIVSEPDIRSPKEAYAYLEKLRSIIQYTGVSDVKMEEGSLRCDANISLRPYGQEKFGTKAELKNLNSFNYVRKGLEYEEKRQEEELLNGGEIGQETRRFDESTGKTILMRVKEGSDDYRYFPEPDIVPLYIDDAWKERVRQTIPELPDERKAKYVNELGLPAYDAHVLTLTKEMSDFFESTIEHGADVKLTSNWLMGGVNEYLNKNQVELLDTKLTPENLAGMIKLIEDGTMSSKIAKKVFPELAAKGGNAKQIMEDNGLVQISDEATLLKFVNEALDNNEQSVEDYKNGKGKAMGFLVGQIMKASKGQANPQLVNQLLKQELDKRLEHHHHHH GatC MTKVTREEVEHIANLARLQISPEETEEMANTLESILDFAKQNDSADTEGVEPTYHVLDLQNVLREDKAIKGIPQELALKNAKETEDGQFKVPTIMNEEDA Open in a separate window caption a8 Macromolecule-production information (Nakamura et al. , 2006 , 2010 ) 2.2.

Techniques: Clone Assay, Expressing, Plasmid Preparation, Sequencing, Construct, Produced

Journal: Methods in enzymology

Article Title: Chemoenzymatic Synthesis and Applications of Prokaryote-Specific UDP-Sugars

doi: 10.1016/bs.mie.2017.06.003

Figure Lengend Snippet: Plasmids used in this manuscript.

Article Snippet: Transform plasmid into freshly-competent BL21 (DE3) RIL cells, growing transformants on LB-agar selection media containing corresponding vector selection antibiotic (see ) and 30 μg/mL CAM. table ft1 table-wrap mode="anchored" t5 Table 1. caption a7 Construct Vector Resistance Reference Addgene ID GST-PglFΔ1–130 ( Cj ) pGEX4T-2 Carb, 50 μg/mL Olivier et al., 2006 89708 His 8 -TEV-PglC ( Ng ) modified pET30b(+) Kan, 30 μμg/mL Hartley et al., 2011 89709 T7-PglD-His 6 ( Cj ) pET24a(+) Kan, 30 μg/mL Morrison et al., 2010 89710 PseB-His 6 ( Cj ) pET-24a(+) Kan, 30 μg/mL this report 89723 His 8 -TEV-PseC ( Cj ) modified pET30b(+) Kan, 30 μg/mL Hartley et al., 2011 89724 His 8 -TEV-PseH ( Cj ) modified pET30b(+) Kan, 30 μg/mL Hartley et al., 2011 89725 T7-WbpB-His 6 ( Pa ) pET-24a(+) Kan, 30 μg/mL Larkin et al., 2009 89711 T7-WbpE-His 6 ( Pa ) pET-24a(+) Kan, 30 μg/mL Larkin et al., 2009 89712 T7-WbpD-His 6 ( Pa ) pET-24a(+) Kan, 30 μg/mL Larkin et al., 2009 89713 T7-AglK-His 6 ( Mv ) pET-24a(+) Kan, 30 μg/mL Larkin et al., 2013 89714 T7-AglC-His 6 ( Mv ) pET-24a(+) Kan, 30 μg/mL Larkin et al., 2013 89726 Open in a separate window Plasmids used in this manuscript.

Techniques: Plasmid Preparation, Modification

Journal:

Article Title: Schistosomiasis-Induced Experimental Pulmonary Hypertension

doi: 10.2353/ajpath.2010.100063

Figure Lengend Snippet: Reagents for Immunoblots

Article Snippet: Immunohistochemistry and immunofluorescence staining on formalin-fixed and paraffin embedded samples of lung tissue obtained at autopsy from two patients who died of schistosomiasis-associated PAH in Brazil was performed using the reagents listed in the . table ft1 table-wrap mode="anchored" t5 caption a7 Immunostain Antigen retrieval Block Primary antibody Secondary antibody Tertiary reagent p-Smad2/3 Citrate buffer 30 minutes in steamer (Vector Laboratories H-3300) 5% Horse serum in PBS 1 hour at room temperature 1/2000 overnight at 4°C (Cell Signaling Technology 3101) 1/100 AF488 Donkey anti-Rabbit (Invitrogen A21206) 1 hour at room temperature None α-Smooth muscle actin Citrate buffer 30 minutes in steamer (Vector Laboratories H-3300) 5% Goat serum in PBS 1 hour at room temperature 1/200 1 hour at room temperature (DakoCytomation M0851) 1/100 AF594 Goat anti-mouse (Invitrogen A11005) 1 hour at room temperature None CD31 Citrate buffer 30 minutes in steamer (Vector Laboratories H-3300) 1.5% Rabbit serum in PBS 1 hour at room temperature 1:500 1 hour at room temperature (antibody courtesy of Lee Lab, Mayo Clinic) 1/100 Biotin-bound rabbit anti-rat (DakoCytomation E0468) 1 hour at room temperature Strepavidin-horseradish peroxidase (Vector Laboratories SA-5704), then DAB 5 minutes, then hematoxylin Open in a separate window Reagents for Immunostains on Human Tissue

Techniques: Blocking Assay, Plasmid Preparation

Journal:

Article Title: Schistosomiasis-Induced Experimental Pulmonary Hypertension

doi: 10.2353/ajpath.2010.100063

Figure Lengend Snippet: Reagents for Immunostains on Mouse Tissue

Article Snippet: Immunohistochemistry and immunofluorescence staining on formalin-fixed and paraffin embedded samples of lung tissue obtained at autopsy from two patients who died of schistosomiasis-associated PAH in Brazil was performed using the reagents listed in the . table ft1 table-wrap mode="anchored" t5 caption a7 Immunostain Antigen retrieval Block Primary antibody Secondary antibody Tertiary reagent p-Smad2/3 Citrate buffer 30 minutes in steamer (Vector Laboratories H-3300) 5% Horse serum in PBS 1 hour at room temperature 1/2000 overnight at 4°C (Cell Signaling Technology 3101) 1/100 AF488 Donkey anti-Rabbit (Invitrogen A21206) 1 hour at room temperature None α-Smooth muscle actin Citrate buffer 30 minutes in steamer (Vector Laboratories H-3300) 5% Goat serum in PBS 1 hour at room temperature 1/200 1 hour at room temperature (DakoCytomation M0851) 1/100 AF594 Goat anti-mouse (Invitrogen A11005) 1 hour at room temperature None CD31 Citrate buffer 30 minutes in steamer (Vector Laboratories H-3300) 1.5% Rabbit serum in PBS 1 hour at room temperature 1:500 1 hour at room temperature (antibody courtesy of Lee Lab, Mayo Clinic) 1/100 Biotin-bound rabbit anti-rat (DakoCytomation E0468) 1 hour at room temperature Strepavidin-horseradish peroxidase (Vector Laboratories SA-5704), then DAB 5 minutes, then hematoxylin Open in a separate window Reagents for Immunostains on Human Tissue

Techniques: Blocking Assay, Plasmid Preparation, Avidin-Biotin Assay, TUNEL Assay

Journal:

Article Title: Schistosomiasis-Induced Experimental Pulmonary Hypertension

doi: 10.2353/ajpath.2010.100063

Figure Lengend Snippet: Reagents for Immunostains on Human Tissue

Article Snippet: Immunohistochemistry and immunofluorescence staining on formalin-fixed and paraffin embedded samples of lung tissue obtained at autopsy from two patients who died of schistosomiasis-associated PAH in Brazil was performed using the reagents listed in the . table ft1 table-wrap mode="anchored" t5 caption a7 Immunostain Antigen retrieval Block Primary antibody Secondary antibody Tertiary reagent p-Smad2/3 Citrate buffer 30 minutes in steamer (Vector Laboratories H-3300) 5% Horse serum in PBS 1 hour at room temperature 1/2000 overnight at 4°C (Cell Signaling Technology 3101) 1/100 AF488 Donkey anti-Rabbit (Invitrogen A21206) 1 hour at room temperature None α-Smooth muscle actin Citrate buffer 30 minutes in steamer (Vector Laboratories H-3300) 5% Goat serum in PBS 1 hour at room temperature 1/200 1 hour at room temperature (DakoCytomation M0851) 1/100 AF594 Goat anti-mouse (Invitrogen A11005) 1 hour at room temperature None CD31 Citrate buffer 30 minutes in steamer (Vector Laboratories H-3300) 1.5% Rabbit serum in PBS 1 hour at room temperature 1:500 1 hour at room temperature (antibody courtesy of Lee Lab, Mayo Clinic) 1/100 Biotin-bound rabbit anti-rat (DakoCytomation E0468) 1 hour at room temperature Strepavidin-horseradish peroxidase (Vector Laboratories SA-5704), then DAB 5 minutes, then hematoxylin Open in a separate window Reagents for Immunostains on Human Tissue

Techniques: Blocking Assay, Plasmid Preparation

Journal:

Article Title: Schistosomiasis-Induced Experimental Pulmonary Hypertension

doi: 10.2353/ajpath.2010.100063

Figure Lengend Snippet: RELM-α and p-Smad2 are up-regulated in response to S. mansoni infection. A: RELM-α, a downstream target of IL-4 and IL-13 signaling, was expressed in pulmonary peri-egg granulomas in infected mice, but suppressed in the absence of IL-13Rα1, as demonstrated by immunostaining (arrowheads mark positive cells) and immunoblot (B; densitometry: five animals per group). There was no evidence of pulmonary intravascular RELM-α expression (representative vessel from wild-type infected animal shown). (The full Western blot is shown in Supplemental Figure 13, see http://ajp.amjpathol.org). C: p-Smad2/3 activity, a downstream target of TGF-β signaling, is increased in both granulomas and vessels with S. mansoni infection (arrowheads mark positive cells), and quantified by immunoblot normalized by total SMAD2 (D; densitometry: three to five animals per group; P = 0.44 by analysis of variance). Only bands for Smad2 (61 kDa) were seen (Scale bars are 50 μm.).

Article Snippet: Immunohistochemistry and immunofluorescence staining on formalin-fixed and paraffin embedded samples of lung tissue obtained at autopsy from two patients who died of schistosomiasis-associated PAH in Brazil was performed using the reagents listed in the . table ft1 table-wrap mode="anchored" t5 caption a7 Immunostain Antigen retrieval Block Primary antibody Secondary antibody Tertiary reagent p-Smad2/3 Citrate buffer 30 minutes in steamer (Vector Laboratories H-3300) 5% Horse serum in PBS 1 hour at room temperature 1/2000 overnight at 4°C (Cell Signaling Technology 3101) 1/100 AF488 Donkey anti-Rabbit (Invitrogen A21206) 1 hour at room temperature None α-Smooth muscle actin Citrate buffer 30 minutes in steamer (Vector Laboratories H-3300) 5% Goat serum in PBS 1 hour at room temperature 1/200 1 hour at room temperature (DakoCytomation M0851) 1/100 AF594 Goat anti-mouse (Invitrogen A11005) 1 hour at room temperature None CD31 Citrate buffer 30 minutes in steamer (Vector Laboratories H-3300) 1.5% Rabbit serum in PBS 1 hour at room temperature 1:500 1 hour at room temperature (antibody courtesy of Lee Lab, Mayo Clinic) 1/100 Biotin-bound rabbit anti-rat (DakoCytomation E0468) 1 hour at room temperature Strepavidin-horseradish peroxidase (Vector Laboratories SA-5704), then DAB 5 minutes, then hematoxylin Open in a separate window Reagents for Immunostains on Human Tissue

Techniques: Infection, Immunostaining, Western Blot, Expressing, Activity Assay

Journal:

Article Title: Schistosomiasis-Induced Experimental Pulmonary Hypertension

doi: 10.2353/ajpath.2010.100063

Figure Lengend Snippet: p-Smad2/3 signaling is present in vascular lesions of patients who died of schistosomiasis-associated PAH. Tissue from two patients was immunostained. A: Costaining for α-smooth muscle actin (αSM-actin) and p-Smad2/3 reveals nuclear activity in the media of vessels with thickened media (arrowheads mark representative positive cells). B: Costaining for CD31 and p-Smad2/3 reveals nuclear activity in many cells present within plexiform lesions (arrowheads mark representative positive cells) (Scale bars: 50 μm.)

Article Snippet: Immunohistochemistry and immunofluorescence staining on formalin-fixed and paraffin embedded samples of lung tissue obtained at autopsy from two patients who died of schistosomiasis-associated PAH in Brazil was performed using the reagents listed in the . table ft1 table-wrap mode="anchored" t5 caption a7 Immunostain Antigen retrieval Block Primary antibody Secondary antibody Tertiary reagent p-Smad2/3 Citrate buffer 30 minutes in steamer (Vector Laboratories H-3300) 5% Horse serum in PBS 1 hour at room temperature 1/2000 overnight at 4°C (Cell Signaling Technology 3101) 1/100 AF488 Donkey anti-Rabbit (Invitrogen A21206) 1 hour at room temperature None α-Smooth muscle actin Citrate buffer 30 minutes in steamer (Vector Laboratories H-3300) 5% Goat serum in PBS 1 hour at room temperature 1/200 1 hour at room temperature (DakoCytomation M0851) 1/100 AF594 Goat anti-mouse (Invitrogen A11005) 1 hour at room temperature None CD31 Citrate buffer 30 minutes in steamer (Vector Laboratories H-3300) 1.5% Rabbit serum in PBS 1 hour at room temperature 1:500 1 hour at room temperature (antibody courtesy of Lee Lab, Mayo Clinic) 1/100 Biotin-bound rabbit anti-rat (DakoCytomation E0468) 1 hour at room temperature Strepavidin-horseradish peroxidase (Vector Laboratories SA-5704), then DAB 5 minutes, then hematoxylin Open in a separate window Reagents for Immunostains on Human Tissue

Techniques: Activity Assay