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psbbigp okt3 tf85g9 bite  (Thermo Fisher)
99
Thermo Fisher psbbigp okt3 tf85g9 bite
Anti-TF-anti-CD3 BiTE activates T cells in the presence of TF-expressing cells in vitro . A Expi293F cells were transfected with pSBbi-GP plasmid containing <t>OKT3-TF85G9</t> BiTE. His-tagged protein was purified from cell culture supernatant via a Nickel-NTA column. OKT3-TF85G9 BiTE protein either not boiled/not reduced (nBnR) or boiled/reduced (BR) were run on an SDS-PAGE gel and Coomassie blue stained. Gel representative of at least two preparations. B HEK293T cells were transiently transfected or co-transfected with plasmids containing either human TF, anti-TF-anti-CD3 BiTE, or an empty vector as stated. At 16 h post-transfection, Jurkat T cells were added and further incubated for 24 h before IL-2 release was analysed by ELISA. Untransfected HEK293T with Jurkat cells or co-transfected TF/BiTE without Jurkat cells were used as additional negative controls, and PMA/ionomycin as a positive control. Bar indicates mean with SD error bars of three pooled independent experiments. One-way ANOVA with Bonferroni post-correction test performed: ns = not significant; * P < 0.05; **** P < 0.0001
Psbbigp Okt3 Tf85g9 Bite, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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restriction cloning  (New England Biolabs)
96
New England Biolabs restriction cloning
Anti-TF-anti-CD3 BiTE activates T cells in the presence of TF-expressing cells in vitro . A Expi293F cells were transfected with pSBbi-GP plasmid containing <t>OKT3-TF85G9</t> BiTE. His-tagged protein was purified from cell culture supernatant via a Nickel-NTA column. OKT3-TF85G9 BiTE protein either not boiled/not reduced (nBnR) or boiled/reduced (BR) were run on an SDS-PAGE gel and Coomassie blue stained. Gel representative of at least two preparations. B HEK293T cells were transiently transfected or co-transfected with plasmids containing either human TF, anti-TF-anti-CD3 BiTE, or an empty vector as stated. At 16 h post-transfection, Jurkat T cells were added and further incubated for 24 h before IL-2 release was analysed by ELISA. Untransfected HEK293T with Jurkat cells or co-transfected TF/BiTE without Jurkat cells were used as additional negative controls, and PMA/ionomycin as a positive control. Bar indicates mean with SD error bars of three pooled independent experiments. One-way ANOVA with Bonferroni post-correction test performed: ns = not significant; * P < 0.05; **** P < 0.0001
Restriction Cloning, supplied by New England Biolabs, 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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psbbigh  (Addgene inc)
92
Addgene inc psbbigh
Anti-TF-anti-CD3 BiTE activates T cells in the presence of TF-expressing cells in vitro . A Expi293F cells were transfected with pSBbi-GP plasmid containing <t>OKT3-TF85G9</t> BiTE. His-tagged protein was purified from cell culture supernatant via a Nickel-NTA column. OKT3-TF85G9 BiTE protein either not boiled/not reduced (nBnR) or boiled/reduced (BR) were run on an SDS-PAGE gel and Coomassie blue stained. Gel representative of at least two preparations. B HEK293T cells were transiently transfected or co-transfected with plasmids containing either human TF, anti-TF-anti-CD3 BiTE, or an empty vector as stated. At 16 h post-transfection, Jurkat T cells were added and further incubated for 24 h before IL-2 release was analysed by ELISA. Untransfected HEK293T with Jurkat cells or co-transfected TF/BiTE without Jurkat cells were used as additional negative controls, and PMA/ionomycin as a positive control. Bar indicates mean with SD error bars of three pooled independent experiments. One-way ANOVA with Bonferroni post-correction test performed: ns = not significant; * P < 0.05; **** P < 0.0001
Psbbigh, supplied by Addgene inc, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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psbbign #60517  (Addgene inc)
90
Addgene inc psbbign #60517
Anti-TF-anti-CD3 BiTE activates T cells in the presence of TF-expressing cells in vitro . A Expi293F cells were transfected with pSBbi-GP plasmid containing <t>OKT3-TF85G9</t> BiTE. His-tagged protein was purified from cell culture supernatant via a Nickel-NTA column. OKT3-TF85G9 BiTE protein either not boiled/not reduced (nBnR) or boiled/reduced (BR) were run on an SDS-PAGE gel and Coomassie blue stained. Gel representative of at least two preparations. B HEK293T cells were transiently transfected or co-transfected with plasmids containing either human TF, anti-TF-anti-CD3 BiTE, or an empty vector as stated. At 16 h post-transfection, Jurkat T cells were added and further incubated for 24 h before IL-2 release was analysed by ELISA. Untransfected HEK293T with Jurkat cells or co-transfected TF/BiTE without Jurkat cells were used as additional negative controls, and PMA/ionomycin as a positive control. Bar indicates mean with SD error bars of three pooled independent experiments. One-way ANOVA with Bonferroni post-correction test performed: ns = not significant; * P < 0.05; **** P < 0.0001
Psbbign #60517, supplied by Addgene inc, 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-TF-anti-CD3 BiTE activates T cells in the presence of TF-expressing cells in vitro . A Expi293F cells were transfected with pSBbi-GP plasmid containing OKT3-TF85G9 BiTE. His-tagged protein was purified from cell culture supernatant via a Nickel-NTA column. OKT3-TF85G9 BiTE protein either not boiled/not reduced (nBnR) or boiled/reduced (BR) were run on an SDS-PAGE gel and Coomassie blue stained. Gel representative of at least two preparations. B HEK293T cells were transiently transfected or co-transfected with plasmids containing either human TF, anti-TF-anti-CD3 BiTE, or an empty vector as stated. At 16 h post-transfection, Jurkat T cells were added and further incubated for 24 h before IL-2 release was analysed by ELISA. Untransfected HEK293T with Jurkat cells or co-transfected TF/BiTE without Jurkat cells were used as additional negative controls, and PMA/ionomycin as a positive control. Bar indicates mean with SD error bars of three pooled independent experiments. One-way ANOVA with Bonferroni post-correction test performed: ns = not significant; * P < 0.05; **** P < 0.0001

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Conversion of anti-tissue factor antibody sequences to chimeric antigen receptor and bi-specific T-cell engager format

doi: 10.1007/s00262-024-03778-3

Figure Lengend Snippet: Anti-TF-anti-CD3 BiTE activates T cells in the presence of TF-expressing cells in vitro . A Expi293F cells were transfected with pSBbi-GP plasmid containing OKT3-TF85G9 BiTE. His-tagged protein was purified from cell culture supernatant via a Nickel-NTA column. OKT3-TF85G9 BiTE protein either not boiled/not reduced (nBnR) or boiled/reduced (BR) were run on an SDS-PAGE gel and Coomassie blue stained. Gel representative of at least two preparations. B HEK293T cells were transiently transfected or co-transfected with plasmids containing either human TF, anti-TF-anti-CD3 BiTE, or an empty vector as stated. At 16 h post-transfection, Jurkat T cells were added and further incubated for 24 h before IL-2 release was analysed by ELISA. Untransfected HEK293T with Jurkat cells or co-transfected TF/BiTE without Jurkat cells were used as additional negative controls, and PMA/ionomycin as a positive control. Bar indicates mean with SD error bars of three pooled independent experiments. One-way ANOVA with Bonferroni post-correction test performed: ns = not significant; * P < 0.05; **** P < 0.0001

Article Snippet: Planktonic Expi293F cells (37.5 × 10 6 in 15 mL) were transfected with 15 µg of pcDNA3.1(-):TF85G9 scFvFc, hATR-5 scFvFc, or human TF-Fc (Genscript), or pSBbiGP:OKT3-TF85G9 BiTE (His-tagged; OKT3 VH-VL and TF85G9 VH-VL joined via glycine-serine linkers; IDT geneblock) using the Gibco Expifectamine293 transfection kit as per the manufacturer’s instructions (Gibco #A14524).

Techniques: Expressing, In Vitro, Transfection, Plasmid Preparation, Purification, Cell Culture, SDS Page, Staining, Incubation, Enzyme-linked Immunosorbent Assay, Positive Control