h7 Search Results


e coli o157 h7 atcc 35150 ehec  (ATCC)
96
ATCC e coli o157 h7 atcc 35150 ehec
E Coli O157 H7 Atcc 35150 Ehec, supplied by ATCC, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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h 7 dihydrochloride  (Tocris)
90
Tocris h 7 dihydrochloride
H 7 Dihydrochloride, supplied by Tocris, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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b7h7fc  (R&D Systems)
91
R&D Systems b7h7fc
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escherichia coli o157 h7  (ATCC)
97
ATCC escherichia coli o157 h7
Bacterial numbers after treating strawberries contaminated with ( a ) Salmonella Thyphimurium and ( b ) Escherichia coli <t>O157:H7.</t> Data points reflect the mean of triplicate measurements ± standard deviation. Different letters indicate significant differences ( p ≤ 0.05).
Escherichia Coli O157 H7, supplied by ATCC, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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e coli o157 h7 atcc  (ATCC)
96
ATCC e coli o157 h7 atcc
Modified Kirby-Bauer results of thin films coated on glass substrates. Antibacterial performance of ZnO thin-film coatings (ZnO-1 thin film coated with ZnO-1 NPs synthesized via N. hansenii B1; ZnO-2 thin film coated with ZnO-2 NPs synthesized via N. hansenii B2) assessed against E. <t>coli</t> <t>O157:H7</t> and S. aureus . Bacterial suspensions were standardized to 0.5 McFarland, spread on nutrient agar plates, and ZnO thin films were placed onto the agar surface. Plates were incubated at 37 °C for 24 h, and antibacterial activity was quantified by measuring inhibition zone diameters (mm). In the figure, different lower-case letters ( a , b ) in the same row and different capital letters (A, B) in the same column on the data table indicate a significant difference ( P < 0.05) between mean values
E Coli O157 H7 Atcc, supplied by ATCC, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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bacteria e coli o157 h7 atcc 43888  (ATCC)
97
ATCC bacteria e coli o157 h7 atcc 43888
Modified Kirby-Bauer results of thin films coated on glass substrates. Antibacterial performance of ZnO thin-film coatings (ZnO-1 thin film coated with ZnO-1 NPs synthesized via N. hansenii B1; ZnO-2 thin film coated with ZnO-2 NPs synthesized via N. hansenii B2) assessed against E. <t>coli</t> <t>O157:H7</t> and S. aureus . Bacterial suspensions were standardized to 0.5 McFarland, spread on nutrient agar plates, and ZnO thin films were placed onto the agar surface. Plates were incubated at 37 °C for 24 h, and antibacterial activity was quantified by measuring inhibition zone diameters (mm). In the figure, different lower-case letters ( a , b ) in the same row and different capital letters (A, B) in the same column on the data table indicate a significant difference ( P < 0.05) between mean values
Bacteria E Coli O157 H7 Atcc 43888, supplied by ATCC, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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43895 e coli o157  (ATCC)
96
ATCC 43895 e coli o157
Modified Kirby-Bauer results of thin films coated on glass substrates. Antibacterial performance of ZnO thin-film coatings (ZnO-1 thin film coated with ZnO-1 NPs synthesized via N. hansenii B1; ZnO-2 thin film coated with ZnO-2 NPs synthesized via N. hansenii B2) assessed against E. <t>coli</t> <t>O157:H7</t> and S. aureus . Bacterial suspensions were standardized to 0.5 McFarland, spread on nutrient agar plates, and ZnO thin films were placed onto the agar surface. Plates were incubated at 37 °C for 24 h, and antibacterial activity was quantified by measuring inhibition zone diameters (mm). In the figure, different lower-case letters ( a , b ) in the same row and different capital letters (A, B) in the same column on the data table indicate a significant difference ( P < 0.05) between mean values
43895 E Coli O157, supplied by ATCC, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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antibody anti human b7h7 af647 r d systems fab80841r m mouse igg  (R&D Systems)
90
R&D Systems antibody anti human b7h7 af647 r d systems fab80841r m mouse igg
(A) Representative contour plots of NK-cell degranulation induced by S2 cells and S2 cells expressing <t>B7H7,</t> CD48, or both, after incubation for 2 hours at an E:T ratio of 1:2. Degranulation was determined by staining for CD56 and CD107a. (B) NK-cell degranulation as in (A) from several donors. Each symbol represents an independent donor, n=5. (C-F) Cytokine production by NK cells after incubation with the indicated S2 cells at 37°C for 6 hours at an E:T ratio of 1:2. A mAb to CD28H (anti-CD28H) was included at 10 μg/ml to block the B7H7-CD28H interaction. Cells were stained for CD56 and CD107a, fixed and permeabilized, and stained for intracellular cytokines. Expression of IFNγ (C), TNFα (D), MIP-1α (E) and MIP-1β (F) in NK cells from multiple donors. Each symbol represents an independent donor, n=7 in (C), n=5 in (D), n=6 in (E), and n= 6 in (F). All data are presented as the mean ± SEM. *P<0.05, **P<0.01, ***P<0.001 (Mann-Whitney test, two-tailed). All experiments have been repeated at least twice.
Antibody Anti Human B7h7 Af647 R D Systems Fab80841r M Mouse Igg, supplied by R&D Systems, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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n diisopropylethylamine dipea  (Chem Impex International)
95
Chem Impex International n diisopropylethylamine dipea
(A) Representative contour plots of NK-cell degranulation induced by S2 cells and S2 cells expressing <t>B7H7,</t> CD48, or both, after incubation for 2 hours at an E:T ratio of 1:2. Degranulation was determined by staining for CD56 and CD107a. (B) NK-cell degranulation as in (A) from several donors. Each symbol represents an independent donor, n=5. (C-F) Cytokine production by NK cells after incubation with the indicated S2 cells at 37°C for 6 hours at an E:T ratio of 1:2. A mAb to CD28H (anti-CD28H) was included at 10 μg/ml to block the B7H7-CD28H interaction. Cells were stained for CD56 and CD107a, fixed and permeabilized, and stained for intracellular cytokines. Expression of IFNγ (C), TNFα (D), MIP-1α (E) and MIP-1β (F) in NK cells from multiple donors. Each symbol represents an independent donor, n=7 in (C), n=5 in (D), n=6 in (E), and n= 6 in (F). All data are presented as the mean ± SEM. *P<0.05, **P<0.01, ***P<0.001 (Mann-Whitney test, two-tailed). All experiments have been repeated at least twice.
N Diisopropylethylamine Dipea, supplied by Chem Impex International, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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mouse igg 1 b159 antibody anti human b7h7 r d systems mab80841 m mouse igg  (R&D Systems)
90
R&D Systems mouse igg 1 b159 antibody anti human b7h7 r d systems mab80841 m mouse igg
Key Resources Table
Mouse Igg 1 B159 Antibody Anti Human B7h7 R D Systems Mab80841 M Mouse Igg, supplied by R&D Systems, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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anti h7 mouse monoclonal antibody mab ina414  (Novus Biologicals)
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Novus Biologicals anti h7 mouse monoclonal antibody mab ina414
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Anti H7 Mouse Monoclonal Antibody Mab Ina414, supplied by Novus Biologicals, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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axl c 20  (Santa Cruz Biotechnology)
94
Santa Cruz Biotechnology axl c 20
Key Resources Table
Axl C 20, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Image Search Results


Bacterial numbers after treating strawberries contaminated with ( a ) Salmonella Thyphimurium and ( b ) Escherichia coli O157:H7. Data points reflect the mean of triplicate measurements ± standard deviation. Different letters indicate significant differences ( p ≤ 0.05).

Journal: Foods

Article Title: Neutral Electrolyzed Water as Sanitizer Solution in Fresh Foods: The Strawberry as a Study Model

doi: 10.3390/foods15050800

Figure Lengend Snippet: Bacterial numbers after treating strawberries contaminated with ( a ) Salmonella Thyphimurium and ( b ) Escherichia coli O157:H7. Data points reflect the mean of triplicate measurements ± standard deviation. Different letters indicate significant differences ( p ≤ 0.05).

Article Snippet: In the present work, the conservative effect of Neutral Electrolyzed Water (NEW) on strawberries was evaluated on contaminated berries with Salmonella Typhimurium ATCC 13311 and Escherichia coli O157:H7.

Techniques: Standard Deviation

Modified Kirby-Bauer results of thin films coated on glass substrates. Antibacterial performance of ZnO thin-film coatings (ZnO-1 thin film coated with ZnO-1 NPs synthesized via N. hansenii B1; ZnO-2 thin film coated with ZnO-2 NPs synthesized via N. hansenii B2) assessed against E. coli O157:H7 and S. aureus . Bacterial suspensions were standardized to 0.5 McFarland, spread on nutrient agar plates, and ZnO thin films were placed onto the agar surface. Plates were incubated at 37 °C for 24 h, and antibacterial activity was quantified by measuring inhibition zone diameters (mm). In the figure, different lower-case letters ( a , b ) in the same row and different capital letters (A, B) in the same column on the data table indicate a significant difference ( P < 0.05) between mean values

Journal: 3 Biotech

Article Title: Thin films coating with ZnO nanoparticles synthesized via acetic acid bacteria: Structural and antibacterial characterization for food applications

doi: 10.1007/s13205-026-04754-7

Figure Lengend Snippet: Modified Kirby-Bauer results of thin films coated on glass substrates. Antibacterial performance of ZnO thin-film coatings (ZnO-1 thin film coated with ZnO-1 NPs synthesized via N. hansenii B1; ZnO-2 thin film coated with ZnO-2 NPs synthesized via N. hansenii B2) assessed against E. coli O157:H7 and S. aureus . Bacterial suspensions were standardized to 0.5 McFarland, spread on nutrient agar plates, and ZnO thin films were placed onto the agar surface. Plates were incubated at 37 °C for 24 h, and antibacterial activity was quantified by measuring inhibition zone diameters (mm). In the figure, different lower-case letters ( a , b ) in the same row and different capital letters (A, B) in the same column on the data table indicate a significant difference ( P < 0.05) between mean values

Article Snippet: The antibacterial activity of ZnO NPs synthesized using supernatants of AAB strains was tested both on E. coli O157:H7 ATCC 35,150 and S. aureus 6538P to represent both Gram-negative and Gram-positive bacteria, respectively.

Techniques: Modification, Synthesized, Incubation, Activity Assay, Inhibition

(A) Representative contour plots of NK-cell degranulation induced by S2 cells and S2 cells expressing B7H7, CD48, or both, after incubation for 2 hours at an E:T ratio of 1:2. Degranulation was determined by staining for CD56 and CD107a. (B) NK-cell degranulation as in (A) from several donors. Each symbol represents an independent donor, n=5. (C-F) Cytokine production by NK cells after incubation with the indicated S2 cells at 37°C for 6 hours at an E:T ratio of 1:2. A mAb to CD28H (anti-CD28H) was included at 10 μg/ml to block the B7H7-CD28H interaction. Cells were stained for CD56 and CD107a, fixed and permeabilized, and stained for intracellular cytokines. Expression of IFNγ (C), TNFα (D), MIP-1α (E) and MIP-1β (F) in NK cells from multiple donors. Each symbol represents an independent donor, n=7 in (C), n=5 in (D), n=6 in (E), and n= 6 in (F). All data are presented as the mean ± SEM. *P<0.05, **P<0.01, ***P<0.001 (Mann-Whitney test, two-tailed). All experiments have been repeated at least twice.

Journal: Cancer immunology research

Article Title: CD28 homolog is a strong activator of natural killer cells for lysis of B7H7 + tumor cells

doi: 10.1158/2326-6066.CIR-18-0733

Figure Lengend Snippet: (A) Representative contour plots of NK-cell degranulation induced by S2 cells and S2 cells expressing B7H7, CD48, or both, after incubation for 2 hours at an E:T ratio of 1:2. Degranulation was determined by staining for CD56 and CD107a. (B) NK-cell degranulation as in (A) from several donors. Each symbol represents an independent donor, n=5. (C-F) Cytokine production by NK cells after incubation with the indicated S2 cells at 37°C for 6 hours at an E:T ratio of 1:2. A mAb to CD28H (anti-CD28H) was included at 10 μg/ml to block the B7H7-CD28H interaction. Cells were stained for CD56 and CD107a, fixed and permeabilized, and stained for intracellular cytokines. Expression of IFNγ (C), TNFα (D), MIP-1α (E) and MIP-1β (F) in NK cells from multiple donors. Each symbol represents an independent donor, n=7 in (C), n=5 in (D), n=6 in (E), and n= 6 in (F). All data are presented as the mean ± SEM. *P<0.05, **P<0.01, ***P<0.001 (Mann-Whitney test, two-tailed). All experiments have been repeated at least twice.

Article Snippet: ​ Reagent type Designation Source Identifier Monoclonal (M) or Polyclonal (P) Host & Sub-class Clone name Antibody Anti-human TMIGD2/CD28H R&D Systems MAB83162 M Mouse IgG 2b 953743 Antibody Anti-human TMIGD2/CD28H AF647 R&D Systems FAB83162R M Mouse IgG 2b 953743 Antibody Anti-human 2B4 BioLegend 329502 M Mouse IgG 1 C1.7 Antibody Anti-human NKG2D R&D Systems MAB139 M Mouse IgG 1 149810 Antibody Anti-human NKp46 BD Biosciences 557847 M Mouse IgG 1 9-E2 Antibody Anti-human CD2 BD Biosciences 555323 M Mouse IgG 1 RPA-2.10 Antibody Anti-human DNAM-1 BD Biosciences 559787 M Mouse IgG 1 DX11 Antibody Anti-human CD20 hIgG1 chimera (Rituximab) Genentech M Antibody Anti-human CD107a PE BD Biosciences 555801 M Mouse IgG 1 H4A3 Antibody Anti-human CD107a FITC BD Biosciences 555800 M Mouse IgG 1 H4A3 Antibody Anti-human CD56 BV421 BD Biosciences 562751 M Mouse IgG 2b NCAM16.2 Antibody Goat anti-mouse F(ab’) 2 PE Jackson ImmunoResearch 115–116-071 P Goat Antibody Anti-human CD16 BD Biosciences 555404 M Mouse IgG 1 3G8 Antibody Anti-human CD56 BD Biosciences 555513 M Mouse IgG 1 B159 Antibody Anti-human B7H7 R&D Systems MAB80841 M Mouse IgG 1 907812 Antibody Anti-human B7H7 AF647 R&D Systems FAB80841R M Mouse IgG 1 907812 Antibody Anti-human HLA-E (3D12) Gift from D. Geraghty (Fred Hutchinson Cancer Research Center, Seattle) M Mouse IgG 1 3D12 Antibody Anti-human NKG2A (Z199) Beckman Coulter IM2751 M Mouse IgG 2b Z199 Antibody Anti-human IFNγ APC BioLegend 506510 M Mouse IgG 1 B27 Antibody Anti-human TNFα BV650 BioLegend 502937 M Mouse IgG 1 MAb11 Antibody Anti-human MIP-1α FITC Invitrogen {"type":"entrez-nucleotide","attrs":{"text":"MA523564","term_id":"1543382917","term_text":"MA523564"}} MA523564 M Mouse IgG 2b 93342 Antibody Anti-human MIP-1β PerCP-Cy5.5 BD Biosciences 560688 M Mouse IgG 1 D21–1351 Antibody Anti-2A peptide Millipore ABS31 P Rabbit Antibody Anti-phosphotyrosine (4G10) Millipore (Upstate) 05–321 M Mouse IgG 2b 4G10 Chemical compound Near-IR Dead Cell Stain Kit Thermo Fisher Scientific {"type":"entrez-nucleotide","attrs":{"text":"L34975","term_id":"522218","term_text":"L34975"}} L34975 Chemical compound PKH67 Green Fluorescent Cell Linker Kit for General Cell Membrane Labeling SIGMA Aldrich PKH67GL Commercial kit In-Fusion HD cloning kit Clontech Laboratories 639648 Commercial kit ToxiLight Non-Destructive Cytotoxicity BioAssay Kit Lonza LT07–117 Commercial kit EasySep Human NK Cell Enrichment Kit STEMCELL Technologies 19055 Open in a separate window Key Resources Table

Techniques: Expressing, Incubation, Staining, Blocking Assay, MANN-WHITNEY, Two Tailed Test

(A) Representative contour plots of NK-cell degranulation induced by the indicated S2 cells in the presence of anti-S2 serum for 2 hours and measured by staining with fluorophore-conjugated CD56 and CD107a mAbs. (B) Degranulation by NK cells as in (A) from multiple donors. Each symbol represents an independent donor, n=7. (C) Staining for CD20 on 221 and Daudi cells. Shaded histograms indicate staining with isotype control. (D, E) Specific lysis of target cells by NK cells at effector to target cell ratio of 5:1 in the presence of 10 μg/ml Rituximab. Each symbol represents an independent donor, n=5 in both (D) and (E). Target cells were 221 (D) and Daudi (E) cell lines, either untransfected or transfected with B7H7. A mAb to CD28H (anti-CD28H) was included at 10 μg/ml to block the B7H7–CD28H interaction. All data are presented as the mean ± SEM. *P<0.05 (Wilcoxon signed-rank test, paired, two-tailed). All experiments have been repeated at least twice.

Journal: Cancer immunology research

Article Title: CD28 homolog is a strong activator of natural killer cells for lysis of B7H7 + tumor cells

doi: 10.1158/2326-6066.CIR-18-0733

Figure Lengend Snippet: (A) Representative contour plots of NK-cell degranulation induced by the indicated S2 cells in the presence of anti-S2 serum for 2 hours and measured by staining with fluorophore-conjugated CD56 and CD107a mAbs. (B) Degranulation by NK cells as in (A) from multiple donors. Each symbol represents an independent donor, n=7. (C) Staining for CD20 on 221 and Daudi cells. Shaded histograms indicate staining with isotype control. (D, E) Specific lysis of target cells by NK cells at effector to target cell ratio of 5:1 in the presence of 10 μg/ml Rituximab. Each symbol represents an independent donor, n=5 in both (D) and (E). Target cells were 221 (D) and Daudi (E) cell lines, either untransfected or transfected with B7H7. A mAb to CD28H (anti-CD28H) was included at 10 μg/ml to block the B7H7–CD28H interaction. All data are presented as the mean ± SEM. *P<0.05 (Wilcoxon signed-rank test, paired, two-tailed). All experiments have been repeated at least twice.

Article Snippet: ​ Reagent type Designation Source Identifier Monoclonal (M) or Polyclonal (P) Host & Sub-class Clone name Antibody Anti-human TMIGD2/CD28H R&D Systems MAB83162 M Mouse IgG 2b 953743 Antibody Anti-human TMIGD2/CD28H AF647 R&D Systems FAB83162R M Mouse IgG 2b 953743 Antibody Anti-human 2B4 BioLegend 329502 M Mouse IgG 1 C1.7 Antibody Anti-human NKG2D R&D Systems MAB139 M Mouse IgG 1 149810 Antibody Anti-human NKp46 BD Biosciences 557847 M Mouse IgG 1 9-E2 Antibody Anti-human CD2 BD Biosciences 555323 M Mouse IgG 1 RPA-2.10 Antibody Anti-human DNAM-1 BD Biosciences 559787 M Mouse IgG 1 DX11 Antibody Anti-human CD20 hIgG1 chimera (Rituximab) Genentech M Antibody Anti-human CD107a PE BD Biosciences 555801 M Mouse IgG 1 H4A3 Antibody Anti-human CD107a FITC BD Biosciences 555800 M Mouse IgG 1 H4A3 Antibody Anti-human CD56 BV421 BD Biosciences 562751 M Mouse IgG 2b NCAM16.2 Antibody Goat anti-mouse F(ab’) 2 PE Jackson ImmunoResearch 115–116-071 P Goat Antibody Anti-human CD16 BD Biosciences 555404 M Mouse IgG 1 3G8 Antibody Anti-human CD56 BD Biosciences 555513 M Mouse IgG 1 B159 Antibody Anti-human B7H7 R&D Systems MAB80841 M Mouse IgG 1 907812 Antibody Anti-human B7H7 AF647 R&D Systems FAB80841R M Mouse IgG 1 907812 Antibody Anti-human HLA-E (3D12) Gift from D. Geraghty (Fred Hutchinson Cancer Research Center, Seattle) M Mouse IgG 1 3D12 Antibody Anti-human NKG2A (Z199) Beckman Coulter IM2751 M Mouse IgG 2b Z199 Antibody Anti-human IFNγ APC BioLegend 506510 M Mouse IgG 1 B27 Antibody Anti-human TNFα BV650 BioLegend 502937 M Mouse IgG 1 MAb11 Antibody Anti-human MIP-1α FITC Invitrogen {"type":"entrez-nucleotide","attrs":{"text":"MA523564","term_id":"1543382917","term_text":"MA523564"}} MA523564 M Mouse IgG 2b 93342 Antibody Anti-human MIP-1β PerCP-Cy5.5 BD Biosciences 560688 M Mouse IgG 1 D21–1351 Antibody Anti-2A peptide Millipore ABS31 P Rabbit Antibody Anti-phosphotyrosine (4G10) Millipore (Upstate) 05–321 M Mouse IgG 2b 4G10 Chemical compound Near-IR Dead Cell Stain Kit Thermo Fisher Scientific {"type":"entrez-nucleotide","attrs":{"text":"L34975","term_id":"522218","term_text":"L34975"}} L34975 Chemical compound PKH67 Green Fluorescent Cell Linker Kit for General Cell Membrane Labeling SIGMA Aldrich PKH67GL Commercial kit In-Fusion HD cloning kit Clontech Laboratories 639648 Commercial kit ToxiLight Non-Destructive Cytotoxicity BioAssay Kit Lonza LT07–117 Commercial kit EasySep Human NK Cell Enrichment Kit STEMCELL Technologies 19055 Open in a separate window Key Resources Table

Techniques: Staining, Control, Lysis, Transfection, Blocking Assay, Two Tailed Test

(A) Tyrosine-phosphorylation of CD28H wild type and the indicated Tyr mutants upon pervanadate treatment. Transfected 293T cells were treated with pervanadate for 10 minutes. Cell lysates were immunoprecipitated with CD28H mAb. Phospho-tyrosine (4G10) and total CD28H (anti-2A) were detected by immunoblots. (B) Lysis of P815 cells by NKL.CD28H cells at various E:T ratios in presence of the indicated mAbs after 6 hours. (C) Lysis of P815 cells by NKL cell transfectants in presence of mAbs to 2B4 and CD28H at the indicated E:T ratios. NKL.CD28H-WT with mAb to 2B4 served as negative control (Ctrl). The graph represents one of two independent experiments. (D) Lysis of 221.B7H7 cells by NKL cell transfectants at the indicated E:T ratios after 6 hours. Untransfected 221 cells served as negative control. The graph represents one of two independent experiments. (E) Immunofluorescence staining with CD28H antibody of Daudi or Daudi–B7H7 cells incubated with NKL cells expressing CD28H wild-type or Y192F mutant. (F) Fold change in fluorescence intensity of CD28H at the immunological synapse (IS), as compared to cell surface regions away from the synapse (non-IS), as in (E). Each dot represents a single cell. Sample size (n) is shown in the figure. Data are obtained and combined from two independent experiments and presented as the mean ± SEM. ****P<0.0001 (Mann-Whitney test, two-tailed). All experiments have been repeated at least twice.

Journal: Cancer immunology research

Article Title: CD28 homolog is a strong activator of natural killer cells for lysis of B7H7 + tumor cells

doi: 10.1158/2326-6066.CIR-18-0733

Figure Lengend Snippet: (A) Tyrosine-phosphorylation of CD28H wild type and the indicated Tyr mutants upon pervanadate treatment. Transfected 293T cells were treated with pervanadate for 10 minutes. Cell lysates were immunoprecipitated with CD28H mAb. Phospho-tyrosine (4G10) and total CD28H (anti-2A) were detected by immunoblots. (B) Lysis of P815 cells by NKL.CD28H cells at various E:T ratios in presence of the indicated mAbs after 6 hours. (C) Lysis of P815 cells by NKL cell transfectants in presence of mAbs to 2B4 and CD28H at the indicated E:T ratios. NKL.CD28H-WT with mAb to 2B4 served as negative control (Ctrl). The graph represents one of two independent experiments. (D) Lysis of 221.B7H7 cells by NKL cell transfectants at the indicated E:T ratios after 6 hours. Untransfected 221 cells served as negative control. The graph represents one of two independent experiments. (E) Immunofluorescence staining with CD28H antibody of Daudi or Daudi–B7H7 cells incubated with NKL cells expressing CD28H wild-type or Y192F mutant. (F) Fold change in fluorescence intensity of CD28H at the immunological synapse (IS), as compared to cell surface regions away from the synapse (non-IS), as in (E). Each dot represents a single cell. Sample size (n) is shown in the figure. Data are obtained and combined from two independent experiments and presented as the mean ± SEM. ****P<0.0001 (Mann-Whitney test, two-tailed). All experiments have been repeated at least twice.

Article Snippet: ​ Reagent type Designation Source Identifier Monoclonal (M) or Polyclonal (P) Host & Sub-class Clone name Antibody Anti-human TMIGD2/CD28H R&D Systems MAB83162 M Mouse IgG 2b 953743 Antibody Anti-human TMIGD2/CD28H AF647 R&D Systems FAB83162R M Mouse IgG 2b 953743 Antibody Anti-human 2B4 BioLegend 329502 M Mouse IgG 1 C1.7 Antibody Anti-human NKG2D R&D Systems MAB139 M Mouse IgG 1 149810 Antibody Anti-human NKp46 BD Biosciences 557847 M Mouse IgG 1 9-E2 Antibody Anti-human CD2 BD Biosciences 555323 M Mouse IgG 1 RPA-2.10 Antibody Anti-human DNAM-1 BD Biosciences 559787 M Mouse IgG 1 DX11 Antibody Anti-human CD20 hIgG1 chimera (Rituximab) Genentech M Antibody Anti-human CD107a PE BD Biosciences 555801 M Mouse IgG 1 H4A3 Antibody Anti-human CD107a FITC BD Biosciences 555800 M Mouse IgG 1 H4A3 Antibody Anti-human CD56 BV421 BD Biosciences 562751 M Mouse IgG 2b NCAM16.2 Antibody Goat anti-mouse F(ab’) 2 PE Jackson ImmunoResearch 115–116-071 P Goat Antibody Anti-human CD16 BD Biosciences 555404 M Mouse IgG 1 3G8 Antibody Anti-human CD56 BD Biosciences 555513 M Mouse IgG 1 B159 Antibody Anti-human B7H7 R&D Systems MAB80841 M Mouse IgG 1 907812 Antibody Anti-human B7H7 AF647 R&D Systems FAB80841R M Mouse IgG 1 907812 Antibody Anti-human HLA-E (3D12) Gift from D. Geraghty (Fred Hutchinson Cancer Research Center, Seattle) M Mouse IgG 1 3D12 Antibody Anti-human NKG2A (Z199) Beckman Coulter IM2751 M Mouse IgG 2b Z199 Antibody Anti-human IFNγ APC BioLegend 506510 M Mouse IgG 1 B27 Antibody Anti-human TNFα BV650 BioLegend 502937 M Mouse IgG 1 MAb11 Antibody Anti-human MIP-1α FITC Invitrogen {"type":"entrez-nucleotide","attrs":{"text":"MA523564","term_id":"1543382917","term_text":"MA523564"}} MA523564 M Mouse IgG 2b 93342 Antibody Anti-human MIP-1β PerCP-Cy5.5 BD Biosciences 560688 M Mouse IgG 1 D21–1351 Antibody Anti-2A peptide Millipore ABS31 P Rabbit Antibody Anti-phosphotyrosine (4G10) Millipore (Upstate) 05–321 M Mouse IgG 2b 4G10 Chemical compound Near-IR Dead Cell Stain Kit Thermo Fisher Scientific {"type":"entrez-nucleotide","attrs":{"text":"L34975","term_id":"522218","term_text":"L34975"}} L34975 Chemical compound PKH67 Green Fluorescent Cell Linker Kit for General Cell Membrane Labeling SIGMA Aldrich PKH67GL Commercial kit In-Fusion HD cloning kit Clontech Laboratories 639648 Commercial kit ToxiLight Non-Destructive Cytotoxicity BioAssay Kit Lonza LT07–117 Commercial kit EasySep Human NK Cell Enrichment Kit STEMCELL Technologies 19055 Open in a separate window Key Resources Table

Techniques: Transfection, Immunoprecipitation, Western Blot, Lysis, Negative Control, Immunofluorescence, Staining, Incubation, Expressing, Mutagenesis, Fluorescence, MANN-WHITNEY, Two Tailed Test

(A) Design and expression of CD28H chimeric antigen receptors in NKL cells. Full-length or cytoplasmic-domain-deleted (ΔCD) CD28H was fused to the cytoplasmic domain of TCRζ and transfected into NKL cells. Cells were stained with fluorophore-conjugated CD28H mAb. Shaded histograms represent staining of untransfected NKL cells. (B, C) Lysis of 221 cells (B) and 221.B7H7 cells (C) by transfected NKL cells at the indicated E:T ratio after 5 hours. (D) Staining for HLA-E and B7H7 in 221.AEH and 221.AEH.B7H7 cells. Shaded histograms represent staining of untransfected 221 cells. (E) Expression of NKG2A on NKL cells determined by immunostaining. Shaded histograms represent staining with IgG control. (F, G) Lysis of 221.AEH cells (F) and 221.AEH.B7H7 cells (G) by transfected NKL cells at the indicated E:T ratios after 5 hours. The graph represents one of two independent experiments. (H) Staining of B7H7 and HLA-E on HDLM-2 tumor cells. Shaded histograms represent staining with isotype controls. (I) Expression of NKG2A on KHYG-1 cells determined by immunostaining (left). Lysis of HDLM-2 tumor cells by KHYG-1 cells after 5 hours, in the presence or absence of a blocking antibody to NKG2A. The graph represents one of two independent experiments. (J) Lysis of HHLA-2+ HLA-E+ HDLM-2 tumor cells by transfected NKL cells at the indicated E:T ratio after 5 hours. The graph represents one of two independent experiments. All experiments have been repeated at least twice.

Journal: Cancer immunology research

Article Title: CD28 homolog is a strong activator of natural killer cells for lysis of B7H7 + tumor cells

doi: 10.1158/2326-6066.CIR-18-0733

Figure Lengend Snippet: (A) Design and expression of CD28H chimeric antigen receptors in NKL cells. Full-length or cytoplasmic-domain-deleted (ΔCD) CD28H was fused to the cytoplasmic domain of TCRζ and transfected into NKL cells. Cells were stained with fluorophore-conjugated CD28H mAb. Shaded histograms represent staining of untransfected NKL cells. (B, C) Lysis of 221 cells (B) and 221.B7H7 cells (C) by transfected NKL cells at the indicated E:T ratio after 5 hours. (D) Staining for HLA-E and B7H7 in 221.AEH and 221.AEH.B7H7 cells. Shaded histograms represent staining of untransfected 221 cells. (E) Expression of NKG2A on NKL cells determined by immunostaining. Shaded histograms represent staining with IgG control. (F, G) Lysis of 221.AEH cells (F) and 221.AEH.B7H7 cells (G) by transfected NKL cells at the indicated E:T ratios after 5 hours. The graph represents one of two independent experiments. (H) Staining of B7H7 and HLA-E on HDLM-2 tumor cells. Shaded histograms represent staining with isotype controls. (I) Expression of NKG2A on KHYG-1 cells determined by immunostaining (left). Lysis of HDLM-2 tumor cells by KHYG-1 cells after 5 hours, in the presence or absence of a blocking antibody to NKG2A. The graph represents one of two independent experiments. (J) Lysis of HHLA-2+ HLA-E+ HDLM-2 tumor cells by transfected NKL cells at the indicated E:T ratio after 5 hours. The graph represents one of two independent experiments. All experiments have been repeated at least twice.

Article Snippet: ​ Reagent type Designation Source Identifier Monoclonal (M) or Polyclonal (P) Host & Sub-class Clone name Antibody Anti-human TMIGD2/CD28H R&D Systems MAB83162 M Mouse IgG 2b 953743 Antibody Anti-human TMIGD2/CD28H AF647 R&D Systems FAB83162R M Mouse IgG 2b 953743 Antibody Anti-human 2B4 BioLegend 329502 M Mouse IgG 1 C1.7 Antibody Anti-human NKG2D R&D Systems MAB139 M Mouse IgG 1 149810 Antibody Anti-human NKp46 BD Biosciences 557847 M Mouse IgG 1 9-E2 Antibody Anti-human CD2 BD Biosciences 555323 M Mouse IgG 1 RPA-2.10 Antibody Anti-human DNAM-1 BD Biosciences 559787 M Mouse IgG 1 DX11 Antibody Anti-human CD20 hIgG1 chimera (Rituximab) Genentech M Antibody Anti-human CD107a PE BD Biosciences 555801 M Mouse IgG 1 H4A3 Antibody Anti-human CD107a FITC BD Biosciences 555800 M Mouse IgG 1 H4A3 Antibody Anti-human CD56 BV421 BD Biosciences 562751 M Mouse IgG 2b NCAM16.2 Antibody Goat anti-mouse F(ab’) 2 PE Jackson ImmunoResearch 115–116-071 P Goat Antibody Anti-human CD16 BD Biosciences 555404 M Mouse IgG 1 3G8 Antibody Anti-human CD56 BD Biosciences 555513 M Mouse IgG 1 B159 Antibody Anti-human B7H7 R&D Systems MAB80841 M Mouse IgG 1 907812 Antibody Anti-human B7H7 AF647 R&D Systems FAB80841R M Mouse IgG 1 907812 Antibody Anti-human HLA-E (3D12) Gift from D. Geraghty (Fred Hutchinson Cancer Research Center, Seattle) M Mouse IgG 1 3D12 Antibody Anti-human NKG2A (Z199) Beckman Coulter IM2751 M Mouse IgG 2b Z199 Antibody Anti-human IFNγ APC BioLegend 506510 M Mouse IgG 1 B27 Antibody Anti-human TNFα BV650 BioLegend 502937 M Mouse IgG 1 MAb11 Antibody Anti-human MIP-1α FITC Invitrogen {"type":"entrez-nucleotide","attrs":{"text":"MA523564","term_id":"1543382917","term_text":"MA523564"}} MA523564 M Mouse IgG 2b 93342 Antibody Anti-human MIP-1β PerCP-Cy5.5 BD Biosciences 560688 M Mouse IgG 1 D21–1351 Antibody Anti-2A peptide Millipore ABS31 P Rabbit Antibody Anti-phosphotyrosine (4G10) Millipore (Upstate) 05–321 M Mouse IgG 2b 4G10 Chemical compound Near-IR Dead Cell Stain Kit Thermo Fisher Scientific {"type":"entrez-nucleotide","attrs":{"text":"L34975","term_id":"522218","term_text":"L34975"}} L34975 Chemical compound PKH67 Green Fluorescent Cell Linker Kit for General Cell Membrane Labeling SIGMA Aldrich PKH67GL Commercial kit In-Fusion HD cloning kit Clontech Laboratories 639648 Commercial kit ToxiLight Non-Destructive Cytotoxicity BioAssay Kit Lonza LT07–117 Commercial kit EasySep Human NK Cell Enrichment Kit STEMCELL Technologies 19055 Open in a separate window Key Resources Table

Techniques: Expressing, Transfection, Staining, Lysis, Immunostaining, Control, Blocking Assay

Key Resources Table

Journal: Cancer immunology research

Article Title: CD28 homolog is a strong activator of natural killer cells for lysis of B7H7 + tumor cells

doi: 10.1158/2326-6066.CIR-18-0733

Figure Lengend Snippet: Key Resources Table

Article Snippet: ​ Reagent type Designation Source Identifier Monoclonal (M) or Polyclonal (P) Host & Sub-class Clone name Antibody Anti-human TMIGD2/CD28H R&D Systems MAB83162 M Mouse IgG 2b 953743 Antibody Anti-human TMIGD2/CD28H AF647 R&D Systems FAB83162R M Mouse IgG 2b 953743 Antibody Anti-human 2B4 BioLegend 329502 M Mouse IgG 1 C1.7 Antibody Anti-human NKG2D R&D Systems MAB139 M Mouse IgG 1 149810 Antibody Anti-human NKp46 BD Biosciences 557847 M Mouse IgG 1 9-E2 Antibody Anti-human CD2 BD Biosciences 555323 M Mouse IgG 1 RPA-2.10 Antibody Anti-human DNAM-1 BD Biosciences 559787 M Mouse IgG 1 DX11 Antibody Anti-human CD20 hIgG1 chimera (Rituximab) Genentech M Antibody Anti-human CD107a PE BD Biosciences 555801 M Mouse IgG 1 H4A3 Antibody Anti-human CD107a FITC BD Biosciences 555800 M Mouse IgG 1 H4A3 Antibody Anti-human CD56 BV421 BD Biosciences 562751 M Mouse IgG 2b NCAM16.2 Antibody Goat anti-mouse F(ab’) 2 PE Jackson ImmunoResearch 115–116-071 P Goat Antibody Anti-human CD16 BD Biosciences 555404 M Mouse IgG 1 3G8 Antibody Anti-human CD56 BD Biosciences 555513 M Mouse IgG 1 B159 Antibody Anti-human B7H7 R&D Systems MAB80841 M Mouse IgG 1 907812 Antibody Anti-human B7H7 AF647 R&D Systems FAB80841R M Mouse IgG 1 907812 Antibody Anti-human HLA-E (3D12) Gift from D. Geraghty (Fred Hutchinson Cancer Research Center, Seattle) M Mouse IgG 1 3D12 Antibody Anti-human NKG2A (Z199) Beckman Coulter IM2751 M Mouse IgG 2b Z199 Antibody Anti-human IFNγ APC BioLegend 506510 M Mouse IgG 1 B27 Antibody Anti-human TNFα BV650 BioLegend 502937 M Mouse IgG 1 MAb11 Antibody Anti-human MIP-1α FITC Invitrogen {"type":"entrez-nucleotide","attrs":{"text":"MA523564","term_id":"1543382917","term_text":"MA523564"}} MA523564 M Mouse IgG 2b 93342 Antibody Anti-human MIP-1β PerCP-Cy5.5 BD Biosciences 560688 M Mouse IgG 1 D21–1351 Antibody Anti-2A peptide Millipore ABS31 P Rabbit Antibody Anti-phosphotyrosine (4G10) Millipore (Upstate) 05–321 M Mouse IgG 2b 4G10 Chemical compound Near-IR Dead Cell Stain Kit Thermo Fisher Scientific {"type":"entrez-nucleotide","attrs":{"text":"L34975","term_id":"522218","term_text":"L34975"}} L34975 Chemical compound PKH67 Green Fluorescent Cell Linker Kit for General Cell Membrane Labeling SIGMA Aldrich PKH67GL Commercial kit In-Fusion HD cloning kit Clontech Laboratories 639648 Commercial kit ToxiLight Non-Destructive Cytotoxicity BioAssay Kit Lonza LT07–117 Commercial kit EasySep Human NK Cell Enrichment Kit STEMCELL Technologies 19055 Open in a separate window Key Resources Table

Techniques: Staining, Membrane, Labeling, Clone Assay, Bioassay

Key Resources Table

Journal: Cancer immunology research

Article Title: CD28 homolog is a strong activator of natural killer cells for lysis of B7H7 + tumor cells

doi: 10.1158/2326-6066.CIR-18-0733

Figure Lengend Snippet: Key Resources Table

Article Snippet: ​ Reagent type Designation Source Identifier Monoclonal (M) or Polyclonal (P) Host & Sub-class Clone name Antibody Anti-human TMIGD2/CD28H R&D Systems MAB83162 M Mouse IgG 2b 953743 Antibody Anti-human TMIGD2/CD28H AF647 R&D Systems FAB83162R M Mouse IgG 2b 953743 Antibody Anti-human 2B4 BioLegend 329502 M Mouse IgG 1 C1.7 Antibody Anti-human NKG2D R&D Systems MAB139 M Mouse IgG 1 149810 Antibody Anti-human NKp46 BD Biosciences 557847 M Mouse IgG 1 9-E2 Antibody Anti-human CD2 BD Biosciences 555323 M Mouse IgG 1 RPA-2.10 Antibody Anti-human DNAM-1 BD Biosciences 559787 M Mouse IgG 1 DX11 Antibody Anti-human CD20 hIgG1 chimera (Rituximab) Genentech M Antibody Anti-human CD107a PE BD Biosciences 555801 M Mouse IgG 1 H4A3 Antibody Anti-human CD107a FITC BD Biosciences 555800 M Mouse IgG 1 H4A3 Antibody Anti-human CD56 BV421 BD Biosciences 562751 M Mouse IgG 2b NCAM16.2 Antibody Goat anti-mouse F(ab’) 2 PE Jackson ImmunoResearch 115–116-071 P Goat Antibody Anti-human CD16 BD Biosciences 555404 M Mouse IgG 1 3G8 Antibody Anti-human CD56 BD Biosciences 555513 M Mouse IgG 1 B159 Antibody Anti-human B7H7 R&D Systems MAB80841 M Mouse IgG 1 907812 Antibody Anti-human B7H7 AF647 R&D Systems FAB80841R M Mouse IgG 1 907812 Antibody Anti-human HLA-E (3D12) Gift from D. Geraghty (Fred Hutchinson Cancer Research Center, Seattle) M Mouse IgG 1 3D12 Antibody Anti-human NKG2A (Z199) Beckman Coulter IM2751 M Mouse IgG 2b Z199 Antibody Anti-human IFNγ APC BioLegend 506510 M Mouse IgG 1 B27 Antibody Anti-human TNFα BV650 BioLegend 502937 M Mouse IgG 1 MAb11 Antibody Anti-human MIP-1α FITC Invitrogen {"type":"entrez-nucleotide","attrs":{"text":"MA523564","term_id":"1543382917","term_text":"MA523564"}} MA523564 M Mouse IgG 2b 93342 Antibody Anti-human MIP-1β PerCP-Cy5.5 BD Biosciences 560688 M Mouse IgG 1 D21–1351 Antibody Anti-2A peptide Millipore ABS31 P Rabbit Antibody Anti-phosphotyrosine (4G10) Millipore (Upstate) 05–321 M Mouse IgG 2b 4G10 Chemical compound Near-IR Dead Cell Stain Kit Thermo Fisher Scientific {"type":"entrez-nucleotide","attrs":{"text":"L34975","term_id":"522218","term_text":"L34975"}} L34975 Chemical compound PKH67 Green Fluorescent Cell Linker Kit for General Cell Membrane Labeling SIGMA Aldrich PKH67GL Commercial kit In-Fusion HD cloning kit Clontech Laboratories 639648 Commercial kit ToxiLight Non-Destructive Cytotoxicity BioAssay Kit Lonza LT07–117 Commercial kit EasySep Human NK Cell Enrichment Kit STEMCELL Technologies 19055 Open in a separate window Key Resources Table

Techniques: Staining, Membrane, Labeling, Clone Assay, Bioassay