monoclonal mouse anti horse igga igg1  (Bio-Rad)

Journal: Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association

Article Title: Impact of processing on the sensitising capacity and cross-reactivity of cow's and camel milk proteins in a Brown Norway rat study.

doi: 10.1016/j.fct.2024.114761

Figure Lengend Snippet: Fig. 3. Impact of processing on the reactivity against cow’s and camel milk. Comparison of cow’s and camel milk specific (A) IgG1 and (B) IgE reactivity for Brown Norway rats ip immunised with either PBS, cow’s milk, enzyme hydrolysed (EH) cow’s milk, heat treated (HT) cow’s milk, camel milk, EH camel milk and HT camel milk in sera at Day 35. Each symbol represents a single rat and horizontal lines indicate median value. Clinical signs by means of (C) ear swelling and (D) upper gastrointestinal symptoms were evaluated before sacrifice by an ear swelling test and an oral challenge, respectively. Each symbol represents a single rat and bars represent median value. Kruskal-Wallis test followed by Dunn’s post-test was applied to compare three groups. Asterisk(s) indicate statistically significant differences to cow’s and camel milk immunised rats, with: ∗= p ≤0.05, ∗∗= p ≤0.01. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: For IgG1 detection, 50 μL/well of horse radish peroxidase (HRP)-labelled mouse-anti-rat IgG1 (3060-05, Southern Biotech, Birmingham, AL, US) diluted 1:20,000 (v/v) in PBS-T was added to the plates and incubated for 1 h at RT.

Techniques: Comparison

Journal: Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association

Article Title: Impact of processing on the sensitising capacity and cross-reactivity of cow's and camel milk proteins in a Brown Norway rat study.

doi: 10.1016/j.fct.2024.114761

Figure Lengend Snippet: Fig. 2. Immunogenicity and sensitising capacity of cow’s and camel milk products. Comparison of inherent (A) immunogenicity and (B) sensitising capacity of cow’s milk, enzyme hydrolysed (EH) cow’s milk, heat treated (HT) cow’s milk, camel milk, EH camel milk and HT camel milk. Specific IgG1 and IgE were analysed in sera from Day 35 against the same products as the Brown Norway rats were ip immunised with. Each symbol represents a single rat and horizontal lines indicate median value. Kruskal-Wallis test followed by Dunn’s post-test was applied to compare three groups and Mann-Whitney test was applied to compare two groups. Asterisk(s) indicate statistically significant differences to cow’s and camel milk immunised rats, respectively, whereas asterisk(s) on a horizontal line indicates statistically significant difference between the given groups, with: ∗= p ≤0.05, ∗∗= p ≤0.01. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: For IgG1 detection, 50 μL/well of horse radish peroxidase (HRP)-labelled mouse-anti-rat IgG1 (3060-05, Southern Biotech, Birmingham, AL, US) diluted 1:20,000 (v/v) in PBS-T was added to the plates and incubated for 1 h at RT.

Techniques: Immunopeptidomics, Comparison, MANN-WHITNEY

Journal: Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association

Article Title: Impact of processing on the sensitising capacity and cross-reactivity of cow's and camel milk proteins in a Brown Norway rat study.

doi: 10.1016/j.fct.2024.114761

Figure Lengend Snippet: Fig. 4. Inhibitory ELISA for assessment of cow’s and camel milk product competitive capacity for binding to IgE raised against processed version of cow’s and camel milk. (A) competitive capacity of cow’s milk for binding to IgG1 raised against cow’s milk. (B) competitive capacity of cow’s milk and enzyme hydrolysed (EH) cow’s milk for binding to IgG1 raised against EH cow’s milk. (C) competitive capacity of cow’s milk and heat treated (HT) cow’s milk for binding to IgG1 raised against HT cow’s milk. (D) competitive capacity of camel milk for binding to IgG1 raised against camel milk. (E) competitive capacity of camel milk and EH camel milk for binding to IgG1 raised against EH camel milk. (F) competitive capacity of camel milk and HT camel milk for binding to IgG1 raised against HT camel milk. Analyses were performed in duplicates with pooled sera from individual groups and repeated three times. Each symbol represents the mean ( ± SD) percentage of inhibition of IgG1 against the inhibitor concentrations. The stippled line indicates the concentration required for 50% inhibition (IC50).

Article Snippet: For IgG1 detection, 50 μL/well of horse radish peroxidase (HRP)-labelled mouse-anti-rat IgG1 (3060-05, Southern Biotech, Birmingham, AL, US) diluted 1:20,000 (v/v) in PBS-T was added to the plates and incubated for 1 h at RT.

Techniques: Enzyme-linked Immunosorbent Assay, Binding Assay, Inhibition, Concentration Assay

Journal: Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association

Article Title: Impact of processing on the sensitising capacity and cross-reactivity of cow's and camel milk proteins in a Brown Norway rat study.

doi: 10.1016/j.fct.2024.114761

Figure Lengend Snippet: Fig. 6. Reactivity of cow’s and camel milk specific antibodies to processed cow’s and camel milk products, respectively. Comparison of impact of enzyme hydrolysis (EH) and heat treatment (HT) on (A) IgG1 and (B) IgE raised against cow’s and camel milk. Each symbol represents a single rat and horizontal lines indicate median value. Kruskal-Wallis test followed by Dunn’s post-test was applied to compare three groups. Asterisk(s) indicate statistically significant differences to cow’s and camel milk-specific IgG1 and IgE, with: ∗∗= p ≤0.01. (C) competitive capacity of cow’s milk, EH cow’s milk and HT cow’s milk for binding to IgG1 raised against cow’s milk. (D) competitive capacity of camel milk, EH camel milk and HT camel milk for binding to IgG1 raised against camel milk. Analyses were performed in duplicates with pooled sera from individual groups and repeated three times. Each symbol represents the mean ( ± SD) percentage of inhibition of IgG1 against the inhibitor concentrations. The stippled line indicates the concentration required for 50% inhibition (IC50).

Article Snippet: For IgG1 detection, 50 μL/well of horse radish peroxidase (HRP)-labelled mouse-anti-rat IgG1 (3060-05, Southern Biotech, Birmingham, AL, US) diluted 1:20,000 (v/v) in PBS-T was added to the plates and incubated for 1 h at RT.

Techniques: Comparison, Binding Assay, Inhibition, Concentration Assay

Journal: Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association

Article Title: Impact of processing on the sensitising capacity and cross-reactivity of cow's and camel milk proteins in a Brown Norway rat study.

doi: 10.1016/j.fct.2024.114761

Figure Lengend Snippet: Fig. 5. Impact of processing on IgG1 protein specificity. (A) reactivity to cow’s milk proteins by IgG1 raised against cow’s milk (diluted 1:2000 (v/v)) (lane 1), enzyme hydrolysed (EH) cow’s milk (diluted 1:100 (v/v)) (lane 2), or heat treated (HT) cow’s milk (diluted 1:100 (v/v)) (lane 3). (B) reactivity to camel milk proteins by IgG1 raised against camel milk (diluted 1:1000 (v/v)) (lane 1), EH camel milk (diluted 1:200 (v/v)) (lane 2), or HT camel milk (diluted 1:1000 (v/v)) (lane 3). Gels were loaded with 5 μg of cow’s or camel milk proteins. M, protein standard (kDa).

Article Snippet: For IgG1 detection, 50 μL/well of horse radish peroxidase (HRP)-labelled mouse-anti-rat IgG1 (3060-05, Southern Biotech, Birmingham, AL, US) diluted 1:20,000 (v/v) in PBS-T was added to the plates and incubated for 1 h at RT.

Techniques:

Journal: Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association

Article Title: Impact of processing on the sensitising capacity and cross-reactivity of cow's and camel milk proteins in a Brown Norway rat study.

doi: 10.1016/j.fct.2024.114761

Figure Lengend Snippet: Fig. 7. Impact of processing on the cross-reactivity between cow’s and camel milk. Comparison of cow’s (A,B) and camel milk (C,D) specific (A,C) IgG1 and (B,D) IgE against either (A,B) camel milk or (C,D) cow’s milk products. Each symbol represents a single rat and horizontal lines indicate median value. Kruskal- Wallis test followed by Dunn’s post-test was applied to compare four groups. Asterisk(s) indicate statistically significant differences to cow’s and camel milk- specific IgG1 and IgE against cow’s and camel milk, respectively, with: *p ≤0.05, **p ≤0.01, ***p ≤0.001, ****p ≤0.0001.

Article Snippet: For IgG1 detection, 50 μL/well of horse radish peroxidase (HRP)-labelled mouse-anti-rat IgG1 (3060-05, Southern Biotech, Birmingham, AL, US) diluted 1:20,000 (v/v) in PBS-T was added to the plates and incubated for 1 h at RT.

Techniques: Comparison

Journal: Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association

Article Title: Impact of processing on the sensitising capacity and cross-reactivity of cow's and camel milk proteins in a Brown Norway rat study.

doi: 10.1016/j.fct.2024.114761

Figure Lengend Snippet: Fig. 8. Cross-competitive capacity of cow’s and camel milk products. Comparison of impact of enzyme hydrolysis (EH) and heat treatment (HT) on cross- reactivity with IgG1 raised against (A) cow’s milk and (B) camel milk. Analyses were performed in duplicates with pooled sera from individual groups and repeated three times. Each symbol represents the mean (± SD) percentage of inhibition of IgG1 against the inhibitor concentrations. The stippled line indicates the concentration required for 50% inhibition (IC50).

Article Snippet: For IgG1 detection, 50 μL/well of horse radish peroxidase (HRP)-labelled mouse-anti-rat IgG1 (3060-05, Southern Biotech, Birmingham, AL, US) diluted 1:20,000 (v/v) in PBS-T was added to the plates and incubated for 1 h at RT.

Techniques: Comparison, Inhibition, Concentration Assay

Journal: Scientific Reports

Article Title: Construction of Vero cell-adapted rabies vaccine strain by five amino acid substitutions in HEP-Flury strain

doi: 10.1038/s41598-024-63337-9

Figure Lengend Snippet: Efficiency of viral growth, entry, and expression of G protein of each recombinant viruses in Vero cells. ( a ) Information on recombinant viruses used in this figure. *: Amino acid sequence identical with rHEP. **: Amino acid positions are numbered based on the mature G protein without signal peptide. ( b ) Vero cells were inoculated with the indicated recombinant viruses ( a ) at a M.O.I. of 0.05 and supernatants were collected every day until 4 d.p.i. Viral titers were determined in MNA cells. ( c ) The total number of focuses was determined by counting the number of focuses stained with the FITC anti-rabies monoclonal globulin in Vero or MNA cells under a fluorescence microscope. The ratio of the number of focuses in Vero to that in MNA was compared. ( d ) The particle titer of each secreted alkaline phosphatase (SEAP)-expressing VSVp stock was determined in MNA cells. Vero cells then were inoculated with two-fold serial dilutions (starting from 150 particles) of VSVp pseudotyped with the HEP or HEP-10V G protein SEAP activity was assessed in culture supernatants and detected by optical density (OD). ( e ) Vero cells were inoculated with each recombinant virus at an M.O.I. of 5 and harvested at 2 d.p.i. The cells were stained with anti-rabies G protein monoclonal antibody (#7-1-9) and FITC-conjugated anti-mouse secondary. After that, cells were fixed with 4% paraformaldehyde, and finally analyzed by BD FACS Canto II flow cytometer (Becton Dickinson and Company; BD, Franklin Lakes, NJ, USA) and Kaluza analysis software Version 2.1 (Beckman Coulter Life Sciences, Indianapolis, IN, USA). Data are presented as the mean and S.D. from three ( b , d , e ) or four ( c ) independent experiments. Significant differences are indicated in the comparison between rHEP and each recombinant virus after application of two-way ANOVA followed by Tukey ( b ), or each virus after application of one-way ANOVA followed by Turkey ( c , e ) or two-way ANOVA followed by Sidaks ( d ) (*: p < 0.05, **: p < 0.01, ***: p < 0.001).

Article Snippet: Cells were then washed twice and reacted for 1 h on ice with the FITC-conjugated Goat Anti-Mouse IgG1 secondary antibodies (1:1600) (FI-2000; Vector Laboratories, Newark, CA, USA).

Techniques: Expressing, Recombinant, Sequencing, Staining, Fluorescence, Microscopy, Activity Assay, Virus, Flow Cytometry, Software, Comparison