Structured Review

Hemacare pbmcs
XIAP overexpression inhibits <t>ADCC</t> response in SUM149 cells through caspase inhibition. ( a ) Percent cell lysis of SUM149 FG9 and wtXIAP cells incubated with cetuximab alone, PBMC alone or cetuximab and IL-2 activated <t>PBMCs</t> for 4 h, n =2–3. ( b ) Schematic of XIAP mutants used in this study. D148A mutation is known to disrupt binding of caspases 3 and 7, whereas W310A disrupts caspase-9 binding. The H467A point mutation abolishes E3 ubiquitin ligase activity. ( c ) Western immunoblot of XIAP expression in cell lines transduced as indicated. FG9 is an empty vector as described in Materials and Methods. ( d ) Caspase activity and ( e ) viability of XIAP variant cell lines treated as indicated. Bars represent fold change in luminescence ( d ) or mean±S.E.M. % cell death ( e ), n =2–3. ( f ) Percent cell lysis of wtXIAP, +DW/AA and +H/A cells incubated with cetuximab alone, PBMC alone or cetuximab and IL-2 activated PBMCs for 4 h. Bars represent mean±S.E.M. calculated percent lysis, n =2–3. * P
Pbmcs, supplied by Hemacare, 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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1) Product Images from "X-linked inhibitor of apoptosis protein mediates tumor cell resistance to antibody-dependent cellular cytotoxicity"

Article Title: X-linked inhibitor of apoptosis protein mediates tumor cell resistance to antibody-dependent cellular cytotoxicity

Journal: Cell Death & Disease

doi: 10.1038/cddis.2015.412

XIAP overexpression inhibits ADCC response in SUM149 cells through caspase inhibition. ( a ) Percent cell lysis of SUM149 FG9 and wtXIAP cells incubated with cetuximab alone, PBMC alone or cetuximab and IL-2 activated PBMCs for 4 h, n =2–3. ( b ) Schematic of XIAP mutants used in this study. D148A mutation is known to disrupt binding of caspases 3 and 7, whereas W310A disrupts caspase-9 binding. The H467A point mutation abolishes E3 ubiquitin ligase activity. ( c ) Western immunoblot of XIAP expression in cell lines transduced as indicated. FG9 is an empty vector as described in Materials and Methods. ( d ) Caspase activity and ( e ) viability of XIAP variant cell lines treated as indicated. Bars represent fold change in luminescence ( d ) or mean±S.E.M. % cell death ( e ), n =2–3. ( f ) Percent cell lysis of wtXIAP, +DW/AA and +H/A cells incubated with cetuximab alone, PBMC alone or cetuximab and IL-2 activated PBMCs for 4 h. Bars represent mean±S.E.M. calculated percent lysis, n =2–3. * P
Figure Legend Snippet: XIAP overexpression inhibits ADCC response in SUM149 cells through caspase inhibition. ( a ) Percent cell lysis of SUM149 FG9 and wtXIAP cells incubated with cetuximab alone, PBMC alone or cetuximab and IL-2 activated PBMCs for 4 h, n =2–3. ( b ) Schematic of XIAP mutants used in this study. D148A mutation is known to disrupt binding of caspases 3 and 7, whereas W310A disrupts caspase-9 binding. The H467A point mutation abolishes E3 ubiquitin ligase activity. ( c ) Western immunoblot of XIAP expression in cell lines transduced as indicated. FG9 is an empty vector as described in Materials and Methods. ( d ) Caspase activity and ( e ) viability of XIAP variant cell lines treated as indicated. Bars represent fold change in luminescence ( d ) or mean±S.E.M. % cell death ( e ), n =2–3. ( f ) Percent cell lysis of wtXIAP, +DW/AA and +H/A cells incubated with cetuximab alone, PBMC alone or cetuximab and IL-2 activated PBMCs for 4 h. Bars represent mean±S.E.M. calculated percent lysis, n =2–3. * P

Techniques Used: Over Expression, Inhibition, Lysis, Incubation, Mutagenesis, Binding Assay, Activity Assay, Western Blot, Expressing, Plasmid Preparation, Variant Assay

ADCC-mediated ROS accumulation is attenuated by exogenous antioxidants and XIAP overexpression. ( a ) Percent cell lysis of SUM149 cells incubated with cetuximab, PBMC alone or the combination in the presence or absence of concanamycin A (CMA), a perforin inhibitor. Bars represent mean±S.E.M. calculated percent lysis, n =2–3. ( b ) Percent cell lysis of SUM149 cells incubated with cetuximab alone or in ADCC conditions in the presence or absence of qVD (pan-caspase inhibitor), MnTBAP (antioxidant) or the combination. Bars represent mean±S.E.M. calculated percent lysis, n =2–3. * P
Figure Legend Snippet: ADCC-mediated ROS accumulation is attenuated by exogenous antioxidants and XIAP overexpression. ( a ) Percent cell lysis of SUM149 cells incubated with cetuximab, PBMC alone or the combination in the presence or absence of concanamycin A (CMA), a perforin inhibitor. Bars represent mean±S.E.M. calculated percent lysis, n =2–3. ( b ) Percent cell lysis of SUM149 cells incubated with cetuximab alone or in ADCC conditions in the presence or absence of qVD (pan-caspase inhibitor), MnTBAP (antioxidant) or the combination. Bars represent mean±S.E.M. calculated percent lysis, n =2–3. * P

Techniques Used: Over Expression, Lysis, Incubation

Related Articles

Isolation:

Article Title: X-linked inhibitor of apoptosis protein mediates tumor cell resistance to antibody-dependent cellular cytotoxicity
Article Snippet: .. ADCC chromium assay Healthy donor PBMCs were isolated from leukapheresis products (HemaCare Corp., Van Nuys, CA, USA) and stored in LN2 until use. .. The day before performing an ADCC assay, PBMCs were thawed and activated with 600 IU/ml recombinant human IL-2 (Prometheus Laboratories Inc., San Diego, CA, USA) in RPMI-1640/10% huAB serum overnight (16–18 h).

Article Title: Direct Detection of T- and B-Memory Lymphocytes by ImmunoSpot® Assays Reveals HCMV Exposure that Serum Antibodies Fail to Identify
Article Snippet: .. These PBMC donors were recruited by Hemacare (Van Nuys, CA, USA) and the PBMC were isolated by leukapheresis at Hemacare using Hemacare IRB’s. .. The PBMC were cryopreserved at CTL (Cleveland, OH, USA), following protocols that maintain full lymphocyte functionality upon thawing [ ], and were stored in liquid nitrogen vapor until testing in ImmunoSpot® assays.

Article Title: High-Throughput GLP-Capable Target Cell Visualization Assay for Measuring Cell-Mediated Cytotoxicity
Article Snippet: .. These donors were recruited by Hemacare (Van Nyus, CA, USA) and their PBMC isolated by leukapheresis using Hemacare IRBs. .. The PBMC were then cryopreserved at CTL and stored in vapor liquid nitrogen until testing [ ].

Article Title: Circulating fibrocytes traffic to the lungs in response to CXCL12 and mediate fibrosis
Article Snippet: .. Briefly, PBMCs were isolated from leukopheresis packs (HemaCare Inc., Woodland Hills, California, USA) using Ficoll density centrifugation. .. Cells isolated in this way were then cultured in DMEM supplemented with 20% FCS, penicillin, streptomycin, and L -glutamine for 7–14 days.

Article Title: How frequently are predicted peptides actually recognized by CD8 cells?
Article Snippet: .. These PBMC donors were recruited by HemaCare (Van Nyus, Ca), and the PBMC were isolated by leucapheresis at HemaCare using HemaCare IRBs. .. The PBMC were cryopreserved at CTL (Cleveland, OH) and stored in vapor liquid nitrogen until testing in an Enzyme-linked immunospot (ELISPOT) assay.

Centrifugation:

Article Title: Circulating fibrocytes traffic to the lungs in response to CXCL12 and mediate fibrosis
Article Snippet: .. Briefly, PBMCs were isolated from leukopheresis packs (HemaCare Inc., Woodland Hills, California, USA) using Ficoll density centrifugation. .. Cells isolated in this way were then cultured in DMEM supplemented with 20% FCS, penicillin, streptomycin, and L -glutamine for 7–14 days.

Gradient Centrifugation:

Article Title: Identification of broad binding class I HLA supertype epitopes to provide universal coverage of influenza A virus
Article Snippet: .. In the initial screens, Class I typed donor PBMCs from leukapheresis were obtained from HemaCare Corporation, Van Nuys, CA, purified from blood using gradient centrifugation with Histopaque-1077 (Sigma, St Louis, MO) and stored in liquid N2 . .. Frozen donor PBMC were thawed and rested overnight in media containing RPMI 5% AB human serum/complete media followed by a seven day expansion of peptide-specific T cells using a pool of approximately 10 peptides, 1 μg/ml final concentration of each peptide.

Chromium Assay:

Article Title: X-linked inhibitor of apoptosis protein mediates tumor cell resistance to antibody-dependent cellular cytotoxicity
Article Snippet: .. ADCC chromium assay Healthy donor PBMCs were isolated from leukapheresis products (HemaCare Corp., Van Nuys, CA, USA) and stored in LN2 until use. .. The day before performing an ADCC assay, PBMCs were thawed and activated with 600 IU/ml recombinant human IL-2 (Prometheus Laboratories Inc., San Diego, CA, USA) in RPMI-1640/10% huAB serum overnight (16–18 h).

Purification:

Article Title: Identification of broad binding class I HLA supertype epitopes to provide universal coverage of influenza A virus
Article Snippet: .. In the initial screens, Class I typed donor PBMCs from leukapheresis were obtained from HemaCare Corporation, Van Nuys, CA, purified from blood using gradient centrifugation with Histopaque-1077 (Sigma, St Louis, MO) and stored in liquid N2 . .. Frozen donor PBMC were thawed and rested overnight in media containing RPMI 5% AB human serum/complete media followed by a seven day expansion of peptide-specific T cells using a pool of approximately 10 peptides, 1 μg/ml final concentration of each peptide.

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    Hemacare pbmcs
    XIAP overexpression inhibits <t>ADCC</t> response in SUM149 cells through caspase inhibition. ( a ) Percent cell lysis of SUM149 FG9 and wtXIAP cells incubated with cetuximab alone, PBMC alone or cetuximab and IL-2 activated <t>PBMCs</t> for 4 h, n =2–3. ( b ) Schematic of XIAP mutants used in this study. D148A mutation is known to disrupt binding of caspases 3 and 7, whereas W310A disrupts caspase-9 binding. The H467A point mutation abolishes E3 ubiquitin ligase activity. ( c ) Western immunoblot of XIAP expression in cell lines transduced as indicated. FG9 is an empty vector as described in Materials and Methods. ( d ) Caspase activity and ( e ) viability of XIAP variant cell lines treated as indicated. Bars represent fold change in luminescence ( d ) or mean±S.E.M. % cell death ( e ), n =2–3. ( f ) Percent cell lysis of wtXIAP, +DW/AA and +H/A cells incubated with cetuximab alone, PBMC alone or cetuximab and IL-2 activated PBMCs for 4 h. Bars represent mean±S.E.M. calculated percent lysis, n =2–3. * P
    Pbmcs, supplied by Hemacare, used in various techniques. Bioz Stars score: 90/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pbmcs/product/Hemacare
    Average 90 stars, based on 7 article reviews
    Price from $9.99 to $1999.99
    pbmcs - by Bioz Stars, 2020-09
    90/100 stars
      Buy from Supplier

    92
    Hemacare human pbmc
    XIAP overexpression inhibits <t>ADCC</t> response in SUM149 cells through caspase inhibition. ( a ) Percent cell lysis of SUM149 FG9 and wtXIAP cells incubated with cetuximab alone, PBMC alone or cetuximab and IL-2 activated <t>PBMCs</t> for 4 h, n =2–3. ( b ) Schematic of XIAP mutants used in this study. D148A mutation is known to disrupt binding of caspases 3 and 7, whereas W310A disrupts caspase-9 binding. The H467A point mutation abolishes E3 ubiquitin ligase activity. ( c ) Western immunoblot of XIAP expression in cell lines transduced as indicated. FG9 is an empty vector as described in Materials and Methods. ( d ) Caspase activity and ( e ) viability of XIAP variant cell lines treated as indicated. Bars represent fold change in luminescence ( d ) or mean±S.E.M. % cell death ( e ), n =2–3. ( f ) Percent cell lysis of wtXIAP, +DW/AA and +H/A cells incubated with cetuximab alone, PBMC alone or cetuximab and IL-2 activated PBMCs for 4 h. Bars represent mean±S.E.M. calculated percent lysis, n =2–3. * P
    Human Pbmc, supplied by Hemacare, used in various techniques. Bioz Stars score: 92/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/human pbmc/product/Hemacare
    Average 92 stars, based on 9 article reviews
    Price from $9.99 to $1999.99
    human pbmc - by Bioz Stars, 2020-09
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    84
    Hemacare healthy donor derived peripheral blood mononuclear cells pbmcs
    XIAP overexpression inhibits <t>ADCC</t> response in SUM149 cells through caspase inhibition. ( a ) Percent cell lysis of SUM149 FG9 and wtXIAP cells incubated with cetuximab alone, PBMC alone or cetuximab and IL-2 activated <t>PBMCs</t> for 4 h, n =2–3. ( b ) Schematic of XIAP mutants used in this study. D148A mutation is known to disrupt binding of caspases 3 and 7, whereas W310A disrupts caspase-9 binding. The H467A point mutation abolishes E3 ubiquitin ligase activity. ( c ) Western immunoblot of XIAP expression in cell lines transduced as indicated. FG9 is an empty vector as described in Materials and Methods. ( d ) Caspase activity and ( e ) viability of XIAP variant cell lines treated as indicated. Bars represent fold change in luminescence ( d ) or mean±S.E.M. % cell death ( e ), n =2–3. ( f ) Percent cell lysis of wtXIAP, +DW/AA and +H/A cells incubated with cetuximab alone, PBMC alone or cetuximab and IL-2 activated PBMCs for 4 h. Bars represent mean±S.E.M. calculated percent lysis, n =2–3. * P
    Healthy Donor Derived Peripheral Blood Mononuclear Cells Pbmcs, supplied by Hemacare, used in various techniques. Bioz Stars score: 84/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    XIAP overexpression inhibits ADCC response in SUM149 cells through caspase inhibition. ( a ) Percent cell lysis of SUM149 FG9 and wtXIAP cells incubated with cetuximab alone, PBMC alone or cetuximab and IL-2 activated PBMCs for 4 h, n =2–3. ( b ) Schematic of XIAP mutants used in this study. D148A mutation is known to disrupt binding of caspases 3 and 7, whereas W310A disrupts caspase-9 binding. The H467A point mutation abolishes E3 ubiquitin ligase activity. ( c ) Western immunoblot of XIAP expression in cell lines transduced as indicated. FG9 is an empty vector as described in Materials and Methods. ( d ) Caspase activity and ( e ) viability of XIAP variant cell lines treated as indicated. Bars represent fold change in luminescence ( d ) or mean±S.E.M. % cell death ( e ), n =2–3. ( f ) Percent cell lysis of wtXIAP, +DW/AA and +H/A cells incubated with cetuximab alone, PBMC alone or cetuximab and IL-2 activated PBMCs for 4 h. Bars represent mean±S.E.M. calculated percent lysis, n =2–3. * P

    Journal: Cell Death & Disease

    Article Title: X-linked inhibitor of apoptosis protein mediates tumor cell resistance to antibody-dependent cellular cytotoxicity

    doi: 10.1038/cddis.2015.412

    Figure Lengend Snippet: XIAP overexpression inhibits ADCC response in SUM149 cells through caspase inhibition. ( a ) Percent cell lysis of SUM149 FG9 and wtXIAP cells incubated with cetuximab alone, PBMC alone or cetuximab and IL-2 activated PBMCs for 4 h, n =2–3. ( b ) Schematic of XIAP mutants used in this study. D148A mutation is known to disrupt binding of caspases 3 and 7, whereas W310A disrupts caspase-9 binding. The H467A point mutation abolishes E3 ubiquitin ligase activity. ( c ) Western immunoblot of XIAP expression in cell lines transduced as indicated. FG9 is an empty vector as described in Materials and Methods. ( d ) Caspase activity and ( e ) viability of XIAP variant cell lines treated as indicated. Bars represent fold change in luminescence ( d ) or mean±S.E.M. % cell death ( e ), n =2–3. ( f ) Percent cell lysis of wtXIAP, +DW/AA and +H/A cells incubated with cetuximab alone, PBMC alone or cetuximab and IL-2 activated PBMCs for 4 h. Bars represent mean±S.E.M. calculated percent lysis, n =2–3. * P

    Article Snippet: ADCC chromium assay Healthy donor PBMCs were isolated from leukapheresis products (HemaCare Corp., Van Nuys, CA, USA) and stored in LN2 until use.

    Techniques: Over Expression, Inhibition, Lysis, Incubation, Mutagenesis, Binding Assay, Activity Assay, Western Blot, Expressing, Plasmid Preparation, Variant Assay

    ADCC-mediated ROS accumulation is attenuated by exogenous antioxidants and XIAP overexpression. ( a ) Percent cell lysis of SUM149 cells incubated with cetuximab, PBMC alone or the combination in the presence or absence of concanamycin A (CMA), a perforin inhibitor. Bars represent mean±S.E.M. calculated percent lysis, n =2–3. ( b ) Percent cell lysis of SUM149 cells incubated with cetuximab alone or in ADCC conditions in the presence or absence of qVD (pan-caspase inhibitor), MnTBAP (antioxidant) or the combination. Bars represent mean±S.E.M. calculated percent lysis, n =2–3. * P

    Journal: Cell Death & Disease

    Article Title: X-linked inhibitor of apoptosis protein mediates tumor cell resistance to antibody-dependent cellular cytotoxicity

    doi: 10.1038/cddis.2015.412

    Figure Lengend Snippet: ADCC-mediated ROS accumulation is attenuated by exogenous antioxidants and XIAP overexpression. ( a ) Percent cell lysis of SUM149 cells incubated with cetuximab, PBMC alone or the combination in the presence or absence of concanamycin A (CMA), a perforin inhibitor. Bars represent mean±S.E.M. calculated percent lysis, n =2–3. ( b ) Percent cell lysis of SUM149 cells incubated with cetuximab alone or in ADCC conditions in the presence or absence of qVD (pan-caspase inhibitor), MnTBAP (antioxidant) or the combination. Bars represent mean±S.E.M. calculated percent lysis, n =2–3. * P

    Article Snippet: ADCC chromium assay Healthy donor PBMCs were isolated from leukapheresis products (HemaCare Corp., Van Nuys, CA, USA) and stored in LN2 until use.

    Techniques: Over Expression, Lysis, Incubation