Journal: Journal of Clinical Microbiology

Article Title: Identification and Expression of Babesia ovis Secreted Antigen 1 and Evaluation of Its Diagnostic Potential in an Enzyme-Linked Immunosorbent Assay

doi: 10.1128/JCM.03219-14

Figure Lengend Snippet: SDS-PAGE analysis of the recombinant BoSA1 protein expressed in E. coli DH5α. Lane M, protein ladder (kDa). The predicted rBoSA1 protein band expressed in cells is indicated. The figure includes five randomly selected clones of E. coli DH5α cells transformed with recombinant plasmid pGEX-4T3/BoSA1. Lanes E1, E2, E3, E4, and E5, E. coli DH5α cells treated with IPTG. Lanes C1, C2, C3, C4, and C5, uninduced control E. coli DH5α cells.

Article Snippet: Six female ICR mice (CLEA, Japan), 6 weeks of age, were intraperitoneally immunized with 100 μg of purified rBoSA1 emulsified with an equal volume of complete Freund's adjuvant (Difco Laboratories).

Techniques: SDS Page, Recombinant, Clone Assay, Transformation Assay, Plasmid Preparation

Journal: Journal of Clinical Microbiology

Article Title: Identification and Expression of Babesia ovis Secreted Antigen 1 and Evaluation of Its Diagnostic Potential in an Enzyme-Linked Immunosorbent Assay

doi: 10.1128/JCM.03219-14

Figure Lengend Snippet: Western blot analysis of recombinant and native BoSA1 proteins. (A) Western blots showing the reactivity of anti-B. ovis antibodies with rBoSA1. The anti-B. ovis antibodies reacted with several components of rBoSA1 between ∼70 and ∼40 kDa but not with the GST protein. (B) Western blots showing the reactivity of anti-rBoSA1 mouse antibodies with native BoSA1 proteins in the erythrocytes and plasma of B. ovis-infected blood. Lane M, protein ladder; lane iRBC, infected erythrocyte lysate extracted with saponin; lane iP, infected plasma proteins precipitated with saturated ammonium sulfate; lane niRBC, noninfected erythrocyte lysate extracted with saponin; lane niP, noninfected plasma proteins precipitated with saturated ammonium sulfate. Comparisons with molecular size markers indicated estimated molecular masses consistent with the monomeric, dimeric, and tetrameric forms of native BoSA1.

Article Snippet: Six female ICR mice (CLEA, Japan), 6 weeks of age, were intraperitoneally immunized with 100 μg of purified rBoSA1 emulsified with an equal volume of complete Freund's adjuvant (Difco Laboratories).

Techniques: Western Blot, Recombinant, Infection

Journal: Journal of Clinical Microbiology

Article Title: Identification and Expression of Babesia ovis Secreted Antigen 1 and Evaluation of Its Diagnostic Potential in an Enzyme-Linked Immunosorbent Assay

doi: 10.1128/JCM.03219-14

Figure Lengend Snippet: Localization of native BoSA1 proteins recognized by anti-rBoSA1 mouse antibodies in confocal laser micrographs. The images were derived from two sections. (A and E) Phase-contrast images of B. ovis merozoites. (B and F) Overlaid images of fluorescent reactivity and red PI staining on the phase-contrast images of the parasites. (C and G) Immunofluorescence images of parasite-specific antigens. (D and H) Propidium iodide staining of B. ovis merozoite nuclei. Bars = 5 µm.

Article Snippet: Six female ICR mice (CLEA, Japan), 6 weeks of age, were intraperitoneally immunized with 100 μg of purified rBoSA1 emulsified with an equal volume of complete Freund's adjuvant (Difco Laboratories).

Techniques: Derivative Assay, Staining, Immunofluorescence

Journal: PeerJ

Article Title: Generation and characterization of mAb 61H9 against junctional adhesion molecule-a with potent antitumor activity

doi: 10.7717/peerj.17088

Figure Lengend Snippet: (A) Schematic drawing of construct containing the sequence of JAM-A followed by a C-terminal human IgG1 Fc tag. (B) SDS-PAGE analysis of purified JAM-A-Fc proteins. 2 μg of purified production were loaded onto 10% SDS gels.

Article Snippet: JAM-A-Fc (1 μg/mL, 100 μL/well) was placed in coated microplates (Corning, Corning, NY, USA), incubated in a 37 °C incubator for 60 min, then removed and washed with PBST three times.

Techniques: Construct, Sequencing, SDS Page, Purification

Journal: PeerJ

Article Title: Generation and characterization of mAb 61H9 against junctional adhesion molecule-a with potent antitumor activity

doi: 10.7717/peerj.17088

Figure Lengend Snippet: (A) Images for the expression of JAM-A were captured by fluorescence microscope. DAPI staining solution is often used to stain nuclei, which can stain blue. Scale bar, 20 μm. (B) The relative fluorescence intensity was examined by immunofluorescence staining ( Unpaired t-test ). ** P < 0.01,*** P < 0.001

Article Snippet: JAM-A-Fc (1 μg/mL, 100 μL/well) was placed in coated microplates (Corning, Corning, NY, USA), incubated in a 37 °C incubator for 60 min, then removed and washed with PBST three times.

Techniques: Expressing, Fluorescence, Microscopy, Staining, Immunofluorescence

Journal: PeerJ

Article Title: Generation and characterization of mAb 61H9 against junctional adhesion molecule-a with potent antitumor activity

doi: 10.7717/peerj.17088

Figure Lengend Snippet: (A) IgG, negative control group. (B) Commercial antibody FITC-anti-JAM-A (AB275688) was incubated separately, positive group 1. (C) Incubated self-made monoclonal antibody 61H9G4 separately, positive group 2. (D) Self-made antibody was first incubated and then commercial antibody was incubated, competitive inhibition binding test group. (E) Quantification of binding antigen sites of 61H9G4 and AB275688 with JAM-A (Unpaired t-test). **** P < 0.0001

Article Snippet: JAM-A-Fc (1 μg/mL, 100 μL/well) was placed in coated microplates (Corning, Corning, NY, USA), incubated in a 37 °C incubator for 60 min, then removed and washed with PBST three times.

Techniques: Negative Control, Incubation, Inhibition, Binding Assay

Journal: PeerJ

Article Title: Generation and characterization of mAb 61H9 against junctional adhesion molecule-a with potent antitumor activity

doi: 10.7717/peerj.17088

Figure Lengend Snippet: (A) Proliferation ability of JAM-A mAb in KYSE30 and KYSE410 was assessed by CCK-8 assay. (B) The cell apoptosis degree of JAM-A mAb in KYSE30 and KYSE410 was analyzed by flow cytometry ( Two-tailed t-test ). (C) The cell cycle of KYSE30 and KYSE410 with or without JAM-A mAb (Two-tailed t-test). ** p < 0.01, *** p < 0.001.

Article Snippet: JAM-A-Fc (1 μg/mL, 100 μL/well) was placed in coated microplates (Corning, Corning, NY, USA), incubated in a 37 °C incubator for 60 min, then removed and washed with PBST three times.

Techniques: CCK-8 Assay, Flow Cytometry, Two Tailed Test

Journal: PeerJ

Article Title: Generation and characterization of mAb 61H9 against junctional adhesion molecule-a with potent antitumor activity

doi: 10.7717/peerj.17088

Figure Lengend Snippet: (A) Scratch-migration assay at 0, 6, 24, and 48 h after adding JAM-A mAb or IgG in ESCC cells. Scale bar, 100 μm (two-tailed t-test). (B) Transwell invasion assays were carried out in KYSE30 and KYSE410 with JAM-A mAb or IgG (two-tailed t-test). Scale bar, 50 μm. * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: JAM-A-Fc (1 μg/mL, 100 μL/well) was placed in coated microplates (Corning, Corning, NY, USA), incubated in a 37 °C incubator for 60 min, then removed and washed with PBST three times.

Techniques: Migration, Two Tailed Test

Journal: PeerJ

Article Title: Generation and characterization of mAb 61H9 against junctional adhesion molecule-a with potent antitumor activity

doi: 10.7717/peerj.17088

Figure Lengend Snippet: (A) Comparison of tumor volume in the control group and JAM-A mAb group. (B) Tumor growth curves, 10 days after cell inoculation, the tumor volume was measured and tumor size was recorded every 3 days (paired t-test). (C) The tumor cell morphology of nude mice tumor tissues in the control and JAM-A mAb group were detected by HE staining. (D) Immunohistochemical staining for the expression of BCL-2 and IκBα in nude mice. (E) Quantification of overall BCL-2 and IκBα staining intensity in control group and anti-JAM-A group. (Two-tailed t-test), * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: JAM-A-Fc (1 μg/mL, 100 μL/well) was placed in coated microplates (Corning, Corning, NY, USA), incubated in a 37 °C incubator for 60 min, then removed and washed with PBST three times.

Techniques: Comparison, Staining, Immunohistochemical staining, Expressing, Two Tailed Test