maxisorp nunc 96 well plates  (Thermo Fisher)


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    Structured Review

    Thermo Fisher maxisorp nunc 96 well plates
    Maxisorp Nunc 96 Well Plates, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 97/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/maxisorp nunc 96 well plates/product/Thermo Fisher
    Average 97 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    maxisorp nunc 96 well plates - by Bioz Stars, 2020-03
    97/100 stars

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    Related Articles

    Binding Assay:

    Article Title: Antigen-Specific Single B Cell Sorting and Monoclonal Antibody Cloning in Guinea Pigs
    Article Snippet: Paragraph title: mAb Binding Analysis by Enzyme-Linked Immunosorbent Assay (ELISA) ... MaxiSorp 96-well plates (Nunc, Thermo Fisher Scientific) were coated with a mouse anti-His tag mAb (R & D Systems, MAB050) at 2 μg/ml in 100 μl of phosphate buffered saline (PBS) at 4°C overnight.

    Article Title: Mini-HA Is Superior to Full Length Hemagglutinin Immunization in Inducing Stem-Specific Antibodies and Protection Against Group 1 Influenza Virus Challenges in Mice
    Article Snippet: .. ELISA To assess HA-specific antibody binding levels in serum samples, recombinant FL HA of H1 A/California/07/09, H1 A/Puerto Rico/8/34, H1 A/Brisbane/59/07, H2 A/Singapore/01/57 or H5 A/Vietnam/1203/04 (94% sequence identity to H5 A/Hong Kong/156/97 were obtained from Protein Sciences Inc., CT, USA and produced on expres SF+ insect cells) were coated at a concentration of 0.5 μg/mL onto Maxisorp 96-well plates (Nunc, Thermo Scientific) O/N at 4°C. .. Plates were washed with PBS (Life Technologies, Paisley, UK) containing 0.05% Tween-20 (Merck Millipore, Darmstadt, Germany) (PBS-T) and subsequently blocked with PBS containing 2% dried skimmed milk (BD, Breda, the Netherlands) for 1 h at RT.

    Article Title: Functionally Active Fc Mutant Antibodies Recognizing Cancer Antigens Generated Rapidly at High Yields
    Article Snippet: Briefly, Maxisorp 96-well plates (Thermo Fischer Scientific) were coated overnight with 0.5 µg/mL mouse anti-human IgG1 antibody (Bio-Rad) in carbonate buffer. .. On the next day, the plate was washed with PBS 0.5% Tween 20 (PBST) and unspecific binding was blocked using 2% skim milk in PBST (blocking buffer).

    Article Title: Matrix-M Adjuvated Seasonal Virosomal Influenza Vaccine Induces Partial Protection in Mice and Ferrets against Avian H5 and H7 Challenge
    Article Snippet: .. HA-and NA-based ELISA To assess HA- and NA-specific binding antibody levels in serum samples, recombinant (rec) HA (Protein Sciences Inc., CT, USA) and recNA of H1N1 A/California/07/09 (produced on HEK293F cells) or recHA or recNA of H5N1 A/Hong Kong/156/97 (Both produced on HEK293F cells) or recH7 A/Netherlands/219/03 (Protein Sciences Inc.) were coated at a concentration of 0.5μg/ml onto Maxisorp 96-well plates (Nunc, Thermo Scientific) O/N at 4°C. .. Plates were washed with PBS (Life Technologies, Paisley, UK) containing 0.05% Tween-20 (Merck Millipore, Darmstadt, Germany) (PBS-T) and subsequently blocked with PBS containing 2% dried skimmed milk (Becton Dickinson, Breda, the Netherlands) for the HA-based ELISA or with PBS containing 2% bovine serum albumin (BSA; Sigma) (PBS/BSA) for the NA-based ELISA for 1 hour at RT.

    Article Title: Leptospiral Endostatin-Like Protein A Is a Bacterial Cell Surface Receptor for Human Plasminogen ▿
    Article Snippet: .. For enzyme-linked immunosorbent assay (ELISA)-based binding assays, wells of Maxisorp 96-well plates (Nalge Nunc, Rochester, NY) were coated overnight with 10 μg/ml human plasminogen (Sigma-Aldrich), 10 μg/ml recombinant LenA, or 10 μg/ml BSA in 50 mM NaCO3 (pH 9.6) at 4°C. ..

    Article Title: Anti-tumor activity of stability-engineered IgG-like bispecific antibodies targeting TRAIL-R2 and LTβR
    Article Snippet: Supernatants were then assayed for binding to LTβR-Fc antigen by DELFIA. .. MaxiSorp 96-well plates (Nalge Nunc, Rochester, NY) were coated with 1 µg/ml LTβR-Fc in 0.1 M sodium carbonate buffer, pH 9.5 overnight at 4°C, and then blocked with DELFIA assay buffer (DAB, 10 mM Tris HCl pH 7.4, 150 mM NaCl, 20 µM EDTA, 0.5% BSA, 0.02% Tween 20, 0.01% NaN3 ) for 1 h with shaking at room temperature and washed 3 times with DAB without BSA (Wash buffer).

    Article Title: Human Sera Collected between 1979 and 2010 Possess Blocking-Antibody Titers to Pandemic GII.4 Noroviruses Isolated over Three Decades
    Article Snippet: Paragraph title: Determination of serum blocking titers against binding of VLPs to pig gastric mucin. ... Briefly, Thermo Scientific MaxiSorp 96-well plates were coated with pig gastric mucin (Sigma-Aldrich; 10 μg/ml in PBS, pH 7.3) for 4 h at room temperature and subsequently blocked overnight at 4°C with PBS containing 5% dry milk and 0.05% Tween.

    Positive Control:

    Article Title: Scarcity or Absence of Humoral Immune Responses in the Plasma and Cervicovaginal Lavage Fluids of Heavily HIV-1-Exposed But Persistently Seronegative Women
    Article Snippet: The gp120-specific antibody in specimens was measured using previously described methods., , Briefly, MaxiSorp 96-well plates (Nalge Nunc) were coated overnight at room temperature with 1 μg/ml ConB gp120. .. Serial dilutions of CVL or plasma specimens and positive control human serum containing HIV-specific IgA or IgG , were placed in the gp120-coated portions.

    Blocking Assay:

    Article Title: Antigen-Specific Single B Cell Sorting and Monoclonal Antibody Cloning in Guinea Pigs
    Article Snippet: MaxiSorp 96-well plates (Nunc, Thermo Fisher Scientific) were coated with a mouse anti-His tag mAb (R & D Systems, MAB050) at 2 μg/ml in 100 μl of phosphate buffered saline (PBS) at 4°C overnight. .. After incubating with blocking buffer (PBS containing 5% FBS/2% non-fat milk) for 1 h at 37°C, 2 μg/ml of BG505 SOSIP trimers were added into each well and incubated for 1 h at room temperature.

    Article Title: Functionally Active Fc Mutant Antibodies Recognizing Cancer Antigens Generated Rapidly at High Yields
    Article Snippet: Briefly, Maxisorp 96-well plates (Thermo Fischer Scientific) were coated overnight with 0.5 µg/mL mouse anti-human IgG1 antibody (Bio-Rad) in carbonate buffer. .. On the next day, the plate was washed with PBS 0.5% Tween 20 (PBST) and unspecific binding was blocked using 2% skim milk in PBST (blocking buffer).

    Article Title: The Immunomodulatory Role of Adjuvants in Vaccines Formulated with the Recombinant Antigens Ov-103 and Ov-RAL-2 against Onchocerca volvulus in Mice
    Article Snippet: Maxisorp 96-well plates (Nunc Nalgene International, Rochester, NY) were coated with 2 μg /ml of Ov- 103 or Ov- RAL-2 in 50 mM Tris-CI coating buffer pH 8.8 overnight 4°C. .. Borate buffer solution (BBS) (0.17 M boric acid, 0.12 M NaCl, 0.5% Tween-20, 0.025% bovine serum albumin, I mM EDTA, pH 8.2) was used to block the plates for 30 min at room temperature.

    Article Title: Human Sera Collected between 1979 and 2010 Possess Blocking-Antibody Titers to Pandemic GII.4 Noroviruses Isolated over Three Decades
    Article Snippet: Paragraph title: Determination of serum blocking titers against binding of VLPs to pig gastric mucin. ... Briefly, Thermo Scientific MaxiSorp 96-well plates were coated with pig gastric mucin (Sigma-Aldrich; 10 μg/ml in PBS, pH 7.3) for 4 h at room temperature and subsequently blocked overnight at 4°C with PBS containing 5% dry milk and 0.05% Tween.

    Enzyme-linked Immunosorbent Assay:

    Article Title: Antigen-Specific Single B Cell Sorting and Monoclonal Antibody Cloning in Guinea Pigs
    Article Snippet: Paragraph title: mAb Binding Analysis by Enzyme-Linked Immunosorbent Assay (ELISA) ... MaxiSorp 96-well plates (Nunc, Thermo Fisher Scientific) were coated with a mouse anti-His tag mAb (R & D Systems, MAB050) at 2 μg/ml in 100 μl of phosphate buffered saline (PBS) at 4°C overnight.

    Article Title: Mini-HA Is Superior to Full Length Hemagglutinin Immunization in Inducing Stem-Specific Antibodies and Protection Against Group 1 Influenza Virus Challenges in Mice
    Article Snippet: .. ELISA To assess HA-specific antibody binding levels in serum samples, recombinant FL HA of H1 A/California/07/09, H1 A/Puerto Rico/8/34, H1 A/Brisbane/59/07, H2 A/Singapore/01/57 or H5 A/Vietnam/1203/04 (94% sequence identity to H5 A/Hong Kong/156/97 were obtained from Protein Sciences Inc., CT, USA and produced on expres SF+ insect cells) were coated at a concentration of 0.5 μg/mL onto Maxisorp 96-well plates (Nunc, Thermo Scientific) O/N at 4°C. .. Plates were washed with PBS (Life Technologies, Paisley, UK) containing 0.05% Tween-20 (Merck Millipore, Darmstadt, Germany) (PBS-T) and subsequently blocked with PBS containing 2% dried skimmed milk (BD, Breda, the Netherlands) for 1 h at RT.

    Article Title: RKIP contributes to IFN\u03b3 synthesis by CD8+ T cells after serial TCR triggering in Systemic Inflammatory Response Syndrome
    Article Snippet: Paragraph title: Enzyme-linked immunosorbent assay (ELISA) ... Briefly, capture antibody (1:250 dilution) was coated overnight on MaxiSorp 96-well plates (Thermo Scientific; Rochester, NY, USA) in coating buffer (0.1 M sodium carbonate or 0.1M sodium phosphate).

    Article Title: Identification of soluble TREM-2 in the cerebrospinal fluid and its association with multiple sclerosis and CNS inflammation
    Article Snippet: Paragraph title: ELISA for soluble human TREM-2 ... Briefly, anti-human TREM-2 monoclonal antibody (mAb) (clone 20G2) (Bouchon et al. , ) was used as capture antibody and coated overnight at 4°C on Maxisorp 96-well plates (Nalge Nunc International, Rochester, NY) at a final concentration of 5 μg/ml in sodium bicarbonate coating buffer (0.015 M Na2 CO3 + 0.035 M NaHCO3 , pH 9.6).

    Article Title: Functionally Active Fc Mutant Antibodies Recognizing Cancer Antigens Generated Rapidly at High Yields
    Article Snippet: Paragraph title: Evaluations of Antibody Yields by ELISA ... Briefly, Maxisorp 96-well plates (Thermo Fischer Scientific) were coated overnight with 0.5 µg/mL mouse anti-human IgG1 antibody (Bio-Rad) in carbonate buffer.

    Article Title: Matrix-M Adjuvated Seasonal Virosomal Influenza Vaccine Induces Partial Protection in Mice and Ferrets against Avian H5 and H7 Challenge
    Article Snippet: .. HA-and NA-based ELISA To assess HA- and NA-specific binding antibody levels in serum samples, recombinant (rec) HA (Protein Sciences Inc., CT, USA) and recNA of H1N1 A/California/07/09 (produced on HEK293F cells) or recHA or recNA of H5N1 A/Hong Kong/156/97 (Both produced on HEK293F cells) or recH7 A/Netherlands/219/03 (Protein Sciences Inc.) were coated at a concentration of 0.5μg/ml onto Maxisorp 96-well plates (Nunc, Thermo Scientific) O/N at 4°C. .. Plates were washed with PBS (Life Technologies, Paisley, UK) containing 0.05% Tween-20 (Merck Millipore, Darmstadt, Germany) (PBS-T) and subsequently blocked with PBS containing 2% dried skimmed milk (Becton Dickinson, Breda, the Netherlands) for the HA-based ELISA or with PBS containing 2% bovine serum albumin (BSA; Sigma) (PBS/BSA) for the NA-based ELISA for 1 hour at RT.

    Article Title: Scarcity or Absence of Humoral Immune Responses in the Plasma and Cervicovaginal Lavage Fluids of Heavily HIV-1-Exposed But Persistently Seronegative Women
    Article Snippet: Paragraph title: ELISA ... The gp120-specific antibody in specimens was measured using previously described methods., , Briefly, MaxiSorp 96-well plates (Nalge Nunc) were coated overnight at room temperature with 1 μg/ml ConB gp120.

    Article Title: Leptospiral Endostatin-Like Protein A Is a Bacterial Cell Surface Receptor for Human Plasminogen ▿
    Article Snippet: .. For enzyme-linked immunosorbent assay (ELISA)-based binding assays, wells of Maxisorp 96-well plates (Nalge Nunc, Rochester, NY) were coated overnight with 10 μg/ml human plasminogen (Sigma-Aldrich), 10 μg/ml recombinant LenA, or 10 μg/ml BSA in 50 mM NaCO3 (pH 9.6) at 4°C. ..

    Article Title: High Affinity Promotes Internalization of Engineered Antibodies Targeting FGFR1
    Article Snippet: Three cycles of panning were performed on MaxiSorp 96-well plates (Thermo Fisher Scientific; Waltham, MA, USA) coated with FGFR1.D1-D2-D3-Fc with counter selection using purified Fc protein. .. To confirm the interaction with FGFR1 bacterial supernatants containing selected scFv proteins were subjected to the monoclonal ELISA and the SPR assay with immobilized FGFR1.D1-D2-D3-Fc, as described in [ ].

    Article Title: Total Body Irradiation Is Permissive for Mesenchymal Stem Cell-Mediated New Bone Formation Following Local Transplantation
    Article Snippet: Paragraph title: SDF-1α and SDF-1β enzyme-linked immunosorbent assay ... The anti-SDF-1 capture antibody (R & D Systems) in sodium bicarbonate buffer pH 9.4 was bound to MaxiSorp™ 96-well plates (Nunc, Thermo Fisher Scientific) overnight.

    Article Title: The Immunomodulatory Role of Adjuvants in Vaccines Formulated with the Recombinant Antigens Ov-103 and Ov-RAL-2 against Onchocerca volvulus in Mice
    Article Snippet: Paragraph title: ELISA ... Maxisorp 96-well plates (Nunc Nalgene International, Rochester, NY) were coated with 2 μg /ml of Ov- 103 or Ov- RAL-2 in 50 mM Tris-CI coating buffer pH 8.8 overnight 4°C.

    Article Title: Human Sera Collected between 1979 and 2010 Possess Blocking-Antibody Titers to Pandemic GII.4 Noroviruses Isolated over Three Decades
    Article Snippet: Briefly, Thermo Scientific MaxiSorp 96-well plates were coated with pig gastric mucin (Sigma-Aldrich; 10 μg/ml in PBS, pH 7.3) for 4 h at room temperature and subsequently blocked overnight at 4°C with PBS containing 5% dry milk and 0.05% Tween. .. The plates were developed using 100 μl/well of Ultra 1-Step Ultra TMB ELISA HRP substrate solution (Thermo Fisher).

    Sandwich ELISA:

    Article Title: Functionally Active Fc Mutant Antibodies Recognizing Cancer Antigens Generated Rapidly at High Yields
    Article Snippet: Evaluations of Antibody Yields by ELISA IgG antibody expression by Expi293F cells was monitored using a sandwich enzyme-linked immunosorbent assay (ELISA). .. Briefly, Maxisorp 96-well plates (Thermo Fischer Scientific) were coated overnight with 0.5 µg/mL mouse anti-human IgG1 antibody (Bio-Rad) in carbonate buffer.

    Incubation:

    Article Title: Antigen-Specific Single B Cell Sorting and Monoclonal Antibody Cloning in Guinea Pigs
    Article Snippet: MaxiSorp 96-well plates (Nunc, Thermo Fisher Scientific) were coated with a mouse anti-His tag mAb (R & D Systems, MAB050) at 2 μg/ml in 100 μl of phosphate buffered saline (PBS) at 4°C overnight. .. After incubating with blocking buffer (PBS containing 5% FBS/2% non-fat milk) for 1 h at 37°C, 2 μg/ml of BG505 SOSIP trimers were added into each well and incubated for 1 h at room temperature.

    Article Title: Mini-HA Is Superior to Full Length Hemagglutinin Immunization in Inducing Stem-Specific Antibodies and Protection Against Group 1 Influenza Virus Challenges in Mice
    Article Snippet: ELISA To assess HA-specific antibody binding levels in serum samples, recombinant FL HA of H1 A/California/07/09, H1 A/Puerto Rico/8/34, H1 A/Brisbane/59/07, H2 A/Singapore/01/57 or H5 A/Vietnam/1203/04 (94% sequence identity to H5 A/Hong Kong/156/97 were obtained from Protein Sciences Inc., CT, USA and produced on expres SF+ insect cells) were coated at a concentration of 0.5 μg/mL onto Maxisorp 96-well plates (Nunc, Thermo Scientific) O/N at 4°C. .. Following a wash with PBS-T, serum of individual mice were added to the plate in duplicate, serially diluted (2-fold, 0.002–2%) and incubated for 1 h at RT.

    Article Title: RKIP contributes to IFN\u03b3 synthesis by CD8+ T cells after serial TCR triggering in Systemic Inflammatory Response Syndrome
    Article Snippet: Briefly, capture antibody (1:250 dilution) was coated overnight on MaxiSorp 96-well plates (Thermo Scientific; Rochester, NY, USA) in coating buffer (0.1 M sodium carbonate or 0.1M sodium phosphate). .. Next, the plates were washed again and incubated with supernatants from overnight cultures or from mouse sera for 2 h at room temperature.

    Article Title: Identification of soluble TREM-2 in the cerebrospinal fluid and its association with multiple sclerosis and CNS inflammation
    Article Snippet: Briefly, anti-human TREM-2 monoclonal antibody (mAb) (clone 20G2) (Bouchon et al. , ) was used as capture antibody and coated overnight at 4°C on Maxisorp 96-well plates (Nalge Nunc International, Rochester, NY) at a final concentration of 5 μg/ml in sodium bicarbonate coating buffer (0.015 M Na2 CO3 + 0.035 M NaHCO3 , pH 9.6). .. Freshly thawed CSF, serum, DC supernatants and standards were incubated in duplicate overnight at 4°C.

    Article Title: Functionally Active Fc Mutant Antibodies Recognizing Cancer Antigens Generated Rapidly at High Yields
    Article Snippet: Briefly, Maxisorp 96-well plates (Thermo Fischer Scientific) were coated overnight with 0.5 µg/mL mouse anti-human IgG1 antibody (Bio-Rad) in carbonate buffer. .. The plates were developed on the next day, following an incubation with anti-human IgG-HRP (Sigma) at a dilution of 1:5,000, using o-Phenylenediamine dihydrochloride (Sigma) as the HRP substrate.

    Article Title: Matrix-M Adjuvated Seasonal Virosomal Influenza Vaccine Induces Partial Protection in Mice and Ferrets against Avian H5 and H7 Challenge
    Article Snippet: HA-and NA-based ELISA To assess HA- and NA-specific binding antibody levels in serum samples, recombinant (rec) HA (Protein Sciences Inc., CT, USA) and recNA of H1N1 A/California/07/09 (produced on HEK293F cells) or recHA or recNA of H5N1 A/Hong Kong/156/97 (Both produced on HEK293F cells) or recH7 A/Netherlands/219/03 (Protein Sciences Inc.) were coated at a concentration of 0.5μg/ml onto Maxisorp 96-well plates (Nunc, Thermo Scientific) O/N at 4°C. .. Following a wash with PBS-T, serum of individual mice (HA ELISA) or serum pools (NA ELISA) were added to the plate in duplicate, serially diluted (2- fold, 0.002–2%) and incubated for 1 hour at RT.

    Article Title: Native Outer Membrane Proteins Protect Mice against Pulmonary Challenge with Virulent Type A Francisella tularensis
    Article Snippet: .. Wells of MaxiSorp 96-well plates (Nalge Nunc, Rochester, NY) were coated with 100 μl of the bacterial suspension by incubation at 37°C for 2 h, followed by overnight incubation at 4°C. .. The plates were washed three times with wash buffer (PBS containing 0.1% Tween 20), blocked at 37°C for 30 min with PBS containing 10% fetal calf serum (HyClone, Logan, UT), washed three times with wash buffer, and stored at 4°C until they were used.

    Article Title: Leptospiral Endostatin-Like Protein A Is a Bacterial Cell Surface Receptor for Human Plasminogen ▿
    Article Snippet: Membranes were washed with TBS-T and incubated for 1 h at room temperature with goat anti-mouse IgG-horseradish peroxidase (HRP; Santa Cruz Biotechnology), diluted 1:20,000 in TBS-T. Membranes were then developed with SuperSignal West Pico enhanced chemiluminescence substrate (Pierce), and bands were visualized with BioMax Light film (Kodak). .. For enzyme-linked immunosorbent assay (ELISA)-based binding assays, wells of Maxisorp 96-well plates (Nalge Nunc, Rochester, NY) were coated overnight with 10 μg/ml human plasminogen (Sigma-Aldrich), 10 μg/ml recombinant LenA, or 10 μg/ml BSA in 50 mM NaCO3 (pH 9.6) at 4°C.

    Article Title: Leptospiral Endostatin-Like Protein A Is a Bacterial Cell Surface Receptor for Human Plasminogen ▿
    Article Snippet: Controls included intact leptospires incubated with 10 μg/ml of BSA. .. Similarly, Maxisorp 96-well plates (Nalge Nunc) were coated overnight with 10 μg/ml recombinant LenA or 10 μg/ml BSA at 4°C.

    Article Title: Total Body Irradiation Is Permissive for Mesenchymal Stem Cell-Mediated New Bone Formation Following Local Transplantation
    Article Snippet: The anti-SDF-1 capture antibody (R & D Systems) in sodium bicarbonate buffer pH 9.4 was bound to MaxiSorp™ 96-well plates (Nunc, Thermo Fisher Scientific) overnight. .. Murine SDF-1α or SDF-1β (PeproTech) standards and samples (1:2 diluted) were incubated for 2 h prior to incubating with the biotinylated anti-SDF-1α and anti-SDF-1β detection antibody (2 h; R & D Systems), respectively.

    Article Title: Anti-tumor activity of stability-engineered IgG-like bispecific antibodies targeting TRAIL-R2 and LTβR
    Article Snippet: MaxiSorp 96-well plates (Nalge Nunc, Rochester, NY) were coated with 1 µg/ml LTβR-Fc in 0.1 M sodium carbonate buffer, pH 9.5 overnight at 4°C, and then blocked with DELFIA assay buffer (DAB, 10 mM Tris HCl pH 7.4, 150 mM NaCl, 20 µM EDTA, 0.5% BSA, 0.02% Tween 20, 0.01% NaN3 ) for 1 h with shaking at room temperature and washed 3 times with DAB without BSA (Wash buffer). .. Test samples diluted in DAB were added to plates in a final volume of 100 µl, incubated for 1 h with shaking at room temperature, and then rinsed with Wash buffer.

    Article Title: Human Sera Collected between 1979 and 2010 Possess Blocking-Antibody Titers to Pandemic GII.4 Noroviruses Isolated over Three Decades
    Article Snippet: Briefly, Thermo Scientific MaxiSorp 96-well plates were coated with pig gastric mucin (Sigma-Aldrich; 10 μg/ml in PBS, pH 7.3) for 4 h at room temperature and subsequently blocked overnight at 4°C with PBS containing 5% dry milk and 0.05% Tween. .. Next, 2-fold serially diluted sera (starting dilution, 1:40) were mixed with an equal volume of VLPs (at concentrations of 0.06 μg/ml for Grimsby and Houston, 0.08 μg/ml for MD145, and 0.3 μg/ml for Sydney VLPs) and incubated for 1 h at room temperature.

    Diffusion-based Assay:

    Article Title: The Immunomodulatory Role of Adjuvants in Vaccines Formulated with the Recombinant Antigens Ov-103 and Ov-RAL-2 against Onchocerca volvulus in Mice
    Article Snippet: ELISA Serum for antigen-specific antibody analyses was collected when mice received challenge infections within diffusion chambers and at the conclusion of the experiment. lgG1, lgG2a, lgG2b, lgG3, IgM and IgE were measured in mice immunized with Ov- 103 with each of the five adjuvants. .. Maxisorp 96-well plates (Nunc Nalgene International, Rochester, NY) were coated with 2 μg /ml of Ov- 103 or Ov- RAL-2 in 50 mM Tris-CI coating buffer pH 8.8 overnight 4°C.

    Expressing:

    Article Title: Functionally Active Fc Mutant Antibodies Recognizing Cancer Antigens Generated Rapidly at High Yields
    Article Snippet: Evaluations of Antibody Yields by ELISA IgG antibody expression by Expi293F cells was monitored using a sandwich enzyme-linked immunosorbent assay (ELISA). .. Briefly, Maxisorp 96-well plates (Thermo Fischer Scientific) were coated overnight with 0.5 µg/mL mouse anti-human IgG1 antibody (Bio-Rad) in carbonate buffer.

    Article Title: Anti-tumor activity of stability-engineered IgG-like bispecific antibodies targeting TRAIL-R2 and LTβR
    Article Snippet: Colonies were grown overnight in SB expression media supplemented with 1% glycine, 1% Triton X-100, 0.02% arabinose, and 50 µg/ml carbenicillin at either 37°C or 32°C. .. MaxiSorp 96-well plates (Nalge Nunc, Rochester, NY) were coated with 1 µg/ml LTβR-Fc in 0.1 M sodium carbonate buffer, pH 9.5 overnight at 4°C, and then blocked with DELFIA assay buffer (DAB, 10 mM Tris HCl pH 7.4, 150 mM NaCl, 20 µM EDTA, 0.5% BSA, 0.02% Tween 20, 0.01% NaN3 ) for 1 h with shaking at room temperature and washed 3 times with DAB without BSA (Wash buffer).

    Derivative Assay:

    Article Title: Antigen-Specific Single B Cell Sorting and Monoclonal Antibody Cloning in Guinea Pigs
    Article Snippet: mAb Binding Analysis by Enzyme-Linked Immunosorbent Assay (ELISA) The binding specificity of the guinea pig mAbs recovered from single B cells was initially tested with the sorting probe, BG505 SOSIP trimer, and V3 peptides derived from BG505 V3 region (CTRPNNNTRKSIRIGPGQAFYATGDIIGDIRQAHC) and JR-FL (CTRPNNNTRKSIHIGPGRAFYTTGEIIGDIRQAHC) V3 region by ELISA assay. .. MaxiSorp 96-well plates (Nunc, Thermo Fisher Scientific) were coated with a mouse anti-His tag mAb (R & D Systems, MAB050) at 2 μg/ml in 100 μl of phosphate buffered saline (PBS) at 4°C overnight.

    Activation Assay:

    Article Title: Leptospiral Endostatin-Like Protein A Is a Bacterial Cell Surface Receptor for Human Plasminogen ▿
    Article Snippet: Paragraph title: Plasminogen activation assay. ... Similarly, Maxisorp 96-well plates (Nalge Nunc) were coated overnight with 10 μg/ml recombinant LenA or 10 μg/ml BSA at 4°C.

    Cell Culture:

    Article Title: Identification of soluble TREM-2 in the cerebrospinal fluid and its association with multiple sclerosis and CNS inflammation
    Article Snippet: Soluble TREM-2 levels were determined by ELISA on all CSF and serum specimens collected during this study, and on cultured DC supernatant fluids. .. Briefly, anti-human TREM-2 monoclonal antibody (mAb) (clone 20G2) (Bouchon et al. , ) was used as capture antibody and coated overnight at 4°C on Maxisorp 96-well plates (Nalge Nunc International, Rochester, NY) at a final concentration of 5 μg/ml in sodium bicarbonate coating buffer (0.015 M Na2 CO3 + 0.035 M NaHCO3 , pH 9.6).

    SDS Page:

    Article Title: Leptospiral Endostatin-Like Protein A Is a Bacterial Cell Surface Receptor for Human Plasminogen ▿
    Article Snippet: Leptospires were washed twice with PBS and then subjected to 12.5% SDS-PAGE and electrotransferred to nitrocellulose membranes. .. For enzyme-linked immunosorbent assay (ELISA)-based binding assays, wells of Maxisorp 96-well plates (Nalge Nunc, Rochester, NY) were coated overnight with 10 μg/ml human plasminogen (Sigma-Aldrich), 10 μg/ml recombinant LenA, or 10 μg/ml BSA in 50 mM NaCO3 (pH 9.6) at 4°C.

    Polymerase Chain Reaction:

    Article Title: Anti-tumor activity of stability-engineered IgG-like bispecific antibodies targeting TRAIL-R2 and LTβR
    Article Snippet: Bacterial supernatants were collected and heat-treated in either PCR strip tubes or 96-well plates (Applied Biosystems, Foster City, CA) for 60–90 min in a thermal cycler (iCycler, Bio-Rad, Gaithersburg, MD). .. MaxiSorp 96-well plates (Nalge Nunc, Rochester, NY) were coated with 1 µg/ml LTβR-Fc in 0.1 M sodium carbonate buffer, pH 9.5 overnight at 4°C, and then blocked with DELFIA assay buffer (DAB, 10 mM Tris HCl pH 7.4, 150 mM NaCl, 20 µM EDTA, 0.5% BSA, 0.02% Tween 20, 0.01% NaN3 ) for 1 h with shaking at room temperature and washed 3 times with DAB without BSA (Wash buffer).

    Recombinant:

    Article Title: Mini-HA Is Superior to Full Length Hemagglutinin Immunization in Inducing Stem-Specific Antibodies and Protection Against Group 1 Influenza Virus Challenges in Mice
    Article Snippet: .. ELISA To assess HA-specific antibody binding levels in serum samples, recombinant FL HA of H1 A/California/07/09, H1 A/Puerto Rico/8/34, H1 A/Brisbane/59/07, H2 A/Singapore/01/57 or H5 A/Vietnam/1203/04 (94% sequence identity to H5 A/Hong Kong/156/97 were obtained from Protein Sciences Inc., CT, USA and produced on expres SF+ insect cells) were coated at a concentration of 0.5 μg/mL onto Maxisorp 96-well plates (Nunc, Thermo Scientific) O/N at 4°C. .. Plates were washed with PBS (Life Technologies, Paisley, UK) containing 0.05% Tween-20 (Merck Millipore, Darmstadt, Germany) (PBS-T) and subsequently blocked with PBS containing 2% dried skimmed milk (BD, Breda, the Netherlands) for 1 h at RT.

    Article Title: Matrix-M Adjuvated Seasonal Virosomal Influenza Vaccine Induces Partial Protection in Mice and Ferrets against Avian H5 and H7 Challenge
    Article Snippet: .. HA-and NA-based ELISA To assess HA- and NA-specific binding antibody levels in serum samples, recombinant (rec) HA (Protein Sciences Inc., CT, USA) and recNA of H1N1 A/California/07/09 (produced on HEK293F cells) or recHA or recNA of H5N1 A/Hong Kong/156/97 (Both produced on HEK293F cells) or recH7 A/Netherlands/219/03 (Protein Sciences Inc.) were coated at a concentration of 0.5μg/ml onto Maxisorp 96-well plates (Nunc, Thermo Scientific) O/N at 4°C. .. Plates were washed with PBS (Life Technologies, Paisley, UK) containing 0.05% Tween-20 (Merck Millipore, Darmstadt, Germany) (PBS-T) and subsequently blocked with PBS containing 2% dried skimmed milk (Becton Dickinson, Breda, the Netherlands) for the HA-based ELISA or with PBS containing 2% bovine serum albumin (BSA; Sigma) (PBS/BSA) for the NA-based ELISA for 1 hour at RT.

    Article Title: Leptospiral Endostatin-Like Protein A Is a Bacterial Cell Surface Receptor for Human Plasminogen ▿
    Article Snippet: .. For enzyme-linked immunosorbent assay (ELISA)-based binding assays, wells of Maxisorp 96-well plates (Nalge Nunc, Rochester, NY) were coated overnight with 10 μg/ml human plasminogen (Sigma-Aldrich), 10 μg/ml recombinant LenA, or 10 μg/ml BSA in 50 mM NaCO3 (pH 9.6) at 4°C. ..

    Article Title: Leptospiral Endostatin-Like Protein A Is a Bacterial Cell Surface Receptor for Human Plasminogen ▿
    Article Snippet: .. Similarly, Maxisorp 96-well plates (Nalge Nunc) were coated overnight with 10 μg/ml recombinant LenA or 10 μg/ml BSA at 4°C. ..

    Mouse Assay:

    Article Title: Mini-HA Is Superior to Full Length Hemagglutinin Immunization in Inducing Stem-Specific Antibodies and Protection Against Group 1 Influenza Virus Challenges in Mice
    Article Snippet: ELISA To assess HA-specific antibody binding levels in serum samples, recombinant FL HA of H1 A/California/07/09, H1 A/Puerto Rico/8/34, H1 A/Brisbane/59/07, H2 A/Singapore/01/57 or H5 A/Vietnam/1203/04 (94% sequence identity to H5 A/Hong Kong/156/97 were obtained from Protein Sciences Inc., CT, USA and produced on expres SF+ insect cells) were coated at a concentration of 0.5 μg/mL onto Maxisorp 96-well plates (Nunc, Thermo Scientific) O/N at 4°C. .. Following a wash with PBS-T, serum of individual mice were added to the plate in duplicate, serially diluted (2-fold, 0.002–2%) and incubated for 1 h at RT.

    Article Title: Matrix-M Adjuvated Seasonal Virosomal Influenza Vaccine Induces Partial Protection in Mice and Ferrets against Avian H5 and H7 Challenge
    Article Snippet: HA-and NA-based ELISA To assess HA- and NA-specific binding antibody levels in serum samples, recombinant (rec) HA (Protein Sciences Inc., CT, USA) and recNA of H1N1 A/California/07/09 (produced on HEK293F cells) or recHA or recNA of H5N1 A/Hong Kong/156/97 (Both produced on HEK293F cells) or recH7 A/Netherlands/219/03 (Protein Sciences Inc.) were coated at a concentration of 0.5μg/ml onto Maxisorp 96-well plates (Nunc, Thermo Scientific) O/N at 4°C. .. Following a wash with PBS-T, serum of individual mice (HA ELISA) or serum pools (NA ELISA) were added to the plate in duplicate, serially diluted (2- fold, 0.002–2%) and incubated for 1 hour at RT.

    Article Title: The Immunomodulatory Role of Adjuvants in Vaccines Formulated with the Recombinant Antigens Ov-103 and Ov-RAL-2 against Onchocerca volvulus in Mice
    Article Snippet: Antigen-specific lgG1, lgG2a and lgG2b responses were measured in mice immunized with Ov- RAL-2 and co-administered Ov- 103/Ov- RAL-2 formulated alone or with either alum, Advax 2 or MF59. .. Maxisorp 96-well plates (Nunc Nalgene International, Rochester, NY) were coated with 2 μg /ml of Ov- 103 or Ov- RAL-2 in 50 mM Tris-CI coating buffer pH 8.8 overnight 4°C.

    Sequencing:

    Article Title: Mini-HA Is Superior to Full Length Hemagglutinin Immunization in Inducing Stem-Specific Antibodies and Protection Against Group 1 Influenza Virus Challenges in Mice
    Article Snippet: .. ELISA To assess HA-specific antibody binding levels in serum samples, recombinant FL HA of H1 A/California/07/09, H1 A/Puerto Rico/8/34, H1 A/Brisbane/59/07, H2 A/Singapore/01/57 or H5 A/Vietnam/1203/04 (94% sequence identity to H5 A/Hong Kong/156/97 were obtained from Protein Sciences Inc., CT, USA and produced on expres SF+ insect cells) were coated at a concentration of 0.5 μg/mL onto Maxisorp 96-well plates (Nunc, Thermo Scientific) O/N at 4°C. .. Plates were washed with PBS (Life Technologies, Paisley, UK) containing 0.05% Tween-20 (Merck Millipore, Darmstadt, Germany) (PBS-T) and subsequently blocked with PBS containing 2% dried skimmed milk (BD, Breda, the Netherlands) for 1 h at RT.

    Purification:

    Article Title: Scarcity or Absence of Humoral Immune Responses in the Plasma and Cervicovaginal Lavage Fluids of Heavily HIV-1-Exposed But Persistently Seronegative Women
    Article Snippet: The gp120-specific antibody in specimens was measured using previously described methods., , Briefly, MaxiSorp 96-well plates (Nalge Nunc) were coated overnight at room temperature with 1 μg/ml ConB gp120. .. The wells for the standard curve were loaded with known amounts of purified colostral IgA (for IgA assays with CVL) or the above-described human Ig reference serum (for plasma IgA and IgG assays).

    Article Title: High Affinity Promotes Internalization of Engineered Antibodies Targeting FGFR1
    Article Snippet: .. Three cycles of panning were performed on MaxiSorp 96-well plates (Thermo Fisher Scientific; Waltham, MA, USA) coated with FGFR1.D1-D2-D3-Fc with counter selection using purified Fc protein. .. To confirm the interaction with FGFR1 bacterial supernatants containing selected scFv proteins were subjected to the monoclonal ELISA and the SPR assay with immobilized FGFR1.D1-D2-D3-Fc, as described in [ ].

    SPR Assay:

    Article Title: High Affinity Promotes Internalization of Engineered Antibodies Targeting FGFR1
    Article Snippet: Three cycles of panning were performed on MaxiSorp 96-well plates (Thermo Fisher Scientific; Waltham, MA, USA) coated with FGFR1.D1-D2-D3-Fc with counter selection using purified Fc protein. .. To confirm the interaction with FGFR1 bacterial supernatants containing selected scFv proteins were subjected to the monoclonal ELISA and the SPR assay with immobilized FGFR1.D1-D2-D3-Fc, as described in [ ].

    Plasmid Preparation:

    Article Title: Anti-tumor activity of stability-engineered IgG-like bispecific antibodies targeting TRAIL-R2 and LTβR
    Article Snippet: Plasmid DNAs encoding wild-type and engineered BHA10 scFvs were used to transform W3110. .. MaxiSorp 96-well plates (Nalge Nunc, Rochester, NY) were coated with 1 µg/ml LTβR-Fc in 0.1 M sodium carbonate buffer, pH 9.5 overnight at 4°C, and then blocked with DELFIA assay buffer (DAB, 10 mM Tris HCl pH 7.4, 150 mM NaCl, 20 µM EDTA, 0.5% BSA, 0.02% Tween 20, 0.01% NaN3 ) for 1 h with shaking at room temperature and washed 3 times with DAB without BSA (Wash buffer).

    Selection:

    Article Title: High Affinity Promotes Internalization of Engineered Antibodies Targeting FGFR1
    Article Snippet: .. Three cycles of panning were performed on MaxiSorp 96-well plates (Thermo Fisher Scientific; Waltham, MA, USA) coated with FGFR1.D1-D2-D3-Fc with counter selection using purified Fc protein. .. To confirm the interaction with FGFR1 bacterial supernatants containing selected scFv proteins were subjected to the monoclonal ELISA and the SPR assay with immobilized FGFR1.D1-D2-D3-Fc, as described in [ ].

    Produced:

    Article Title: Mini-HA Is Superior to Full Length Hemagglutinin Immunization in Inducing Stem-Specific Antibodies and Protection Against Group 1 Influenza Virus Challenges in Mice
    Article Snippet: .. ELISA To assess HA-specific antibody binding levels in serum samples, recombinant FL HA of H1 A/California/07/09, H1 A/Puerto Rico/8/34, H1 A/Brisbane/59/07, H2 A/Singapore/01/57 or H5 A/Vietnam/1203/04 (94% sequence identity to H5 A/Hong Kong/156/97 were obtained from Protein Sciences Inc., CT, USA and produced on expres SF+ insect cells) were coated at a concentration of 0.5 μg/mL onto Maxisorp 96-well plates (Nunc, Thermo Scientific) O/N at 4°C. .. Plates were washed with PBS (Life Technologies, Paisley, UK) containing 0.05% Tween-20 (Merck Millipore, Darmstadt, Germany) (PBS-T) and subsequently blocked with PBS containing 2% dried skimmed milk (BD, Breda, the Netherlands) for 1 h at RT.

    Article Title: Matrix-M Adjuvated Seasonal Virosomal Influenza Vaccine Induces Partial Protection in Mice and Ferrets against Avian H5 and H7 Challenge
    Article Snippet: .. HA-and NA-based ELISA To assess HA- and NA-specific binding antibody levels in serum samples, recombinant (rec) HA (Protein Sciences Inc., CT, USA) and recNA of H1N1 A/California/07/09 (produced on HEK293F cells) or recHA or recNA of H5N1 A/Hong Kong/156/97 (Both produced on HEK293F cells) or recH7 A/Netherlands/219/03 (Protein Sciences Inc.) were coated at a concentration of 0.5μg/ml onto Maxisorp 96-well plates (Nunc, Thermo Scientific) O/N at 4°C. .. Plates were washed with PBS (Life Technologies, Paisley, UK) containing 0.05% Tween-20 (Merck Millipore, Darmstadt, Germany) (PBS-T) and subsequently blocked with PBS containing 2% dried skimmed milk (Becton Dickinson, Breda, the Netherlands) for the HA-based ELISA or with PBS containing 2% bovine serum albumin (BSA; Sigma) (PBS/BSA) for the NA-based ELISA for 1 hour at RT.

    Concentration Assay:

    Article Title: Mini-HA Is Superior to Full Length Hemagglutinin Immunization in Inducing Stem-Specific Antibodies and Protection Against Group 1 Influenza Virus Challenges in Mice
    Article Snippet: .. ELISA To assess HA-specific antibody binding levels in serum samples, recombinant FL HA of H1 A/California/07/09, H1 A/Puerto Rico/8/34, H1 A/Brisbane/59/07, H2 A/Singapore/01/57 or H5 A/Vietnam/1203/04 (94% sequence identity to H5 A/Hong Kong/156/97 were obtained from Protein Sciences Inc., CT, USA and produced on expres SF+ insect cells) were coated at a concentration of 0.5 μg/mL onto Maxisorp 96-well plates (Nunc, Thermo Scientific) O/N at 4°C. .. Plates were washed with PBS (Life Technologies, Paisley, UK) containing 0.05% Tween-20 (Merck Millipore, Darmstadt, Germany) (PBS-T) and subsequently blocked with PBS containing 2% dried skimmed milk (BD, Breda, the Netherlands) for 1 h at RT.

    Article Title: Identification of soluble TREM-2 in the cerebrospinal fluid and its association with multiple sclerosis and CNS inflammation
    Article Snippet: .. Briefly, anti-human TREM-2 monoclonal antibody (mAb) (clone 20G2) (Bouchon et al. , ) was used as capture antibody and coated overnight at 4°C on Maxisorp 96-well plates (Nalge Nunc International, Rochester, NY) at a final concentration of 5 μg/ml in sodium bicarbonate coating buffer (0.015 M Na2 CO3 + 0.035 M NaHCO3 , pH 9.6). ..

    Article Title: Functionally Active Fc Mutant Antibodies Recognizing Cancer Antigens Generated Rapidly at High Yields
    Article Snippet: Briefly, Maxisorp 96-well plates (Thermo Fischer Scientific) were coated overnight with 0.5 µg/mL mouse anti-human IgG1 antibody (Bio-Rad) in carbonate buffer. .. Trastuzumab (Herceptin® , Roche) was used at as a standard at a starting concentration of 3 µg/mL.

    Article Title: Matrix-M Adjuvated Seasonal Virosomal Influenza Vaccine Induces Partial Protection in Mice and Ferrets against Avian H5 and H7 Challenge
    Article Snippet: .. HA-and NA-based ELISA To assess HA- and NA-specific binding antibody levels in serum samples, recombinant (rec) HA (Protein Sciences Inc., CT, USA) and recNA of H1N1 A/California/07/09 (produced on HEK293F cells) or recHA or recNA of H5N1 A/Hong Kong/156/97 (Both produced on HEK293F cells) or recH7 A/Netherlands/219/03 (Protein Sciences Inc.) were coated at a concentration of 0.5μg/ml onto Maxisorp 96-well plates (Nunc, Thermo Scientific) O/N at 4°C. .. Plates were washed with PBS (Life Technologies, Paisley, UK) containing 0.05% Tween-20 (Merck Millipore, Darmstadt, Germany) (PBS-T) and subsequently blocked with PBS containing 2% dried skimmed milk (Becton Dickinson, Breda, the Netherlands) for the HA-based ELISA or with PBS containing 2% bovine serum albumin (BSA; Sigma) (PBS/BSA) for the NA-based ELISA for 1 hour at RT.

    Article Title: Leptospiral Endostatin-Like Protein A Is a Bacterial Cell Surface Receptor for Human Plasminogen ▿
    Article Snippet: Fifty microliters of resuspended bacteria was transferred to each assay well of a microtiter plate, and 4 ng/well uPA (Chemicon, Temecula, CA) as well as the plasmin-specific substrate d -valyl-leucyl-lysine- p -nitroanilide dihydrochloride (final concentration, 0.3 mM; Sigma-Aldrich) was added. .. Similarly, Maxisorp 96-well plates (Nalge Nunc) were coated overnight with 10 μg/ml recombinant LenA or 10 μg/ml BSA at 4°C.

    Stripping Membranes:

    Article Title: Anti-tumor activity of stability-engineered IgG-like bispecific antibodies targeting TRAIL-R2 and LTβR
    Article Snippet: Bacterial supernatants were collected and heat-treated in either PCR strip tubes or 96-well plates (Applied Biosystems, Foster City, CA) for 60–90 min in a thermal cycler (iCycler, Bio-Rad, Gaithersburg, MD). .. MaxiSorp 96-well plates (Nalge Nunc, Rochester, NY) were coated with 1 µg/ml LTβR-Fc in 0.1 M sodium carbonate buffer, pH 9.5 overnight at 4°C, and then blocked with DELFIA assay buffer (DAB, 10 mM Tris HCl pH 7.4, 150 mM NaCl, 20 µM EDTA, 0.5% BSA, 0.02% Tween 20, 0.01% NaN3 ) for 1 h with shaking at room temperature and washed 3 times with DAB without BSA (Wash buffer).

    Lysis:

    Article Title: Total Body Irradiation Is Permissive for Mesenchymal Stem Cell-Mediated New Bone Formation Following Local Transplantation
    Article Snippet: Briefly, whole BM cell lysates were prepared in Complete Lysis-M EDTA-free buffer containing protease inhibitors (Roche Diagnostics). .. The anti-SDF-1 capture antibody (R & D Systems) in sodium bicarbonate buffer pH 9.4 was bound to MaxiSorp™ 96-well plates (Nunc, Thermo Fisher Scientific) overnight.

    Variant Assay:

    Article Title: Total Body Irradiation Is Permissive for Mesenchymal Stem Cell-Mediated New Bone Formation Following Local Transplantation
    Article Snippet: SDF-1 splice variant-specific enzyme-linked immunosorbent assay (ELISAs; R & D Systems) of whole BM cells and BM interstitial fluids were performed as described previously. .. The anti-SDF-1 capture antibody (R & D Systems) in sodium bicarbonate buffer pH 9.4 was bound to MaxiSorp™ 96-well plates (Nunc, Thermo Fisher Scientific) overnight.

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  • 99
    Thermo Fisher 96 well elisa plates
    Specificity of <t>ELISA</t> and RT–PCR assays The specificity of the PTX3 ELISA was tested using different dilutions of 2C3 antibody to measure immobilized murine and human PTX3. Purified recombinant murine and human PTX3 were immobilized in <t>96‐well</t> ELISA plates, and then, different dilutions of 2C3 were added. The graph shows dose–response of 2C3 on immobilized murine or human PTX3. Human PTX3 was not detected by 2C3 antibody. Evaluation of the amplification efficiency of real‐time RT–PCR assay designed for PTX3 expression in astrocyte cell cultures. (B, C) Melting curve and amplification plot of PTX3 RT–qPCR assay. (D) Standard curves of PTX3 and GAPDH, used as reference mRNA, obtained using fivefold serial dilutions of the cDNA (420, 84, 16.8, 3.36 ng). The threshold cycle ( C t ) values ( y ‐axis) are plotted against log 10 values of cDNA input amounts ( x ‐axis). The graphs are parallel lines and the calculated efficiencies (E) are, respectively, of 1.13 and 1.12 from a y‐slope of −3.04 and −3.07 and a correlation coefficient ( R 2 ) > 0.9.
    96 Well Elisa Plates, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 101 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/96 well elisa plates/product/Thermo Fisher
    Average 99 stars, based on 101 article reviews
    Price from $9.99 to $1999.99
    96 well elisa plates - by Bioz Stars, 2020-03
    99/100 stars
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    99
    Thermo Fisher elisa plates
    Description and characterization of the chimeric human <t>FasL-derived</t> constructs Panel A: Schematic representation of soluble FasL (sFasL), Flag-tagged sFasL (sfFasL), polymeric Flag-tagged soluble FasL (pfFasL), polymeric TCR γ4 and δ5 Flag-tagged soluble FasL generating the TCR-pfFasL upon cotransfection, and beta2-microglobulin-fused HLA-A*02: 01 Flag-tagged soluble FasL (HLA-pfFasL). The f and p symbols represent the flag epitope and the LIF receptor-derived domain triggering the polymerisation of the FasL oligomers, respectively. Panel B: direct immunoblot of the supernatants from COS cells transfected with the empty vector (control) or the FasL constructs sFasL, sfFasL and pfFasL. Panel C: immunoprecipitation of the TCR-pfFasL chimera from transfected HEK cells, using an irrelevant IgG1 antibody, the anti-Flag (clone M2), the anti-FasL (clone 10F2), the anti-TCRγδ (clone IMU-510) or the anti-TCRδ5 (clone 12C7) antibodies. Panel D: immunoprecipitation of the HLA-pfFasL chimera from the supernatant of COS cells, with anti-Flag, anti-FasL or anti-β2microglobulin antibodies. As controls, the same experiment was performed with irrelevant IgG1 and IgG2 antibodies. Panel E: cytotoxic effect of the FasL chimeras. The indicated chimeras, as supernatants from transfected cells and quantitated using the <t>ELISA</t> for FasL, were incubated at the indicated concentrations with Jurkat cells. After 18 h, the MTT cell viability assay was performed. The anti-Flag M2 antibody at 0.5 µg/ml was added to sfFasL to render it cytotoxic.
    Elisa Plates, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 430 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/elisa plates/product/Thermo Fisher
    Average 99 stars, based on 430 article reviews
    Price from $9.99 to $1999.99
    elisa plates - by Bioz Stars, 2020-03
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    78
    Thermo Fisher grp78 elisa microtiter 96 well plates
    Analysis of <t>GRP78–HTJ1</t> and OxPL–GRP78 interactions. (A) GRP78 and HTJ1 expression in HPAEC and human lung microvascular endothelial cells was detected by Western Blot. (B, C) GRP78 interactions were analyzed in coimmunoprecipitation assays using lysates from control or DMPC- (10 μg/ml, 15 min) or OxPAPC-stimulated (10 μg/ml) cells with antibody to GRP78 (B, top), HTJ1 (B, bottom), or EO6 antibody recognizing OxPL (C). (D) Human recombinant GRP78 was incubated with OxPAPC, OxPAPS, or their oxidation-resistant analogues DMPC or DMPS. Left, native gel electrophoresis, followed by Western blot with anti-GRP78 antibody. Shift in electrophoretic mobility of GRP78 incubated with OxPAPS, but not DMPS, indicates formation of GRP78–OxPAPS complex. Right, SDS–PAGE, followed by Western blot with EO6 antibody and reprobing with anti-GRP78 antibody. Positive EO6 immunoreactivity of GRP78 preincubated with OxPAPC indicates formation of GRP78–OxPAPC complex. (E) <t>ELISA</t> plates coated with OxPAPS or DMPS or control uncoated plates incubated with PBS incubated with human recombinant GRP78 (left) or HPAEC lysates (middle and right). The bound GRP78 was detected using anti-GRP78 antibody. * p
    Grp78 Elisa Microtiter 96 Well Plates, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 78/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    grp78 elisa microtiter 96 well plates - by Bioz Stars, 2020-03
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    Specificity of ELISA and RT–PCR assays The specificity of the PTX3 ELISA was tested using different dilutions of 2C3 antibody to measure immobilized murine and human PTX3. Purified recombinant murine and human PTX3 were immobilized in 96‐well ELISA plates, and then, different dilutions of 2C3 were added. The graph shows dose–response of 2C3 on immobilized murine or human PTX3. Human PTX3 was not detected by 2C3 antibody. Evaluation of the amplification efficiency of real‐time RT–PCR assay designed for PTX3 expression in astrocyte cell cultures. (B, C) Melting curve and amplification plot of PTX3 RT–qPCR assay. (D) Standard curves of PTX3 and GAPDH, used as reference mRNA, obtained using fivefold serial dilutions of the cDNA (420, 84, 16.8, 3.36 ng). The threshold cycle ( C t ) values ( y ‐axis) are plotted against log 10 values of cDNA input amounts ( x ‐axis). The graphs are parallel lines and the calculated efficiencies (E) are, respectively, of 1.13 and 1.12 from a y‐slope of −3.04 and −3.07 and a correlation coefficient ( R 2 ) > 0.9.

    Journal: The EMBO Journal

    Article Title: Pentraxin 3 regulates synaptic function by inducing AMPA receptor clustering via ECM remodeling and β1‐integrin

    doi: 10.15252/embj.201899529

    Figure Lengend Snippet: Specificity of ELISA and RT–PCR assays The specificity of the PTX3 ELISA was tested using different dilutions of 2C3 antibody to measure immobilized murine and human PTX3. Purified recombinant murine and human PTX3 were immobilized in 96‐well ELISA plates, and then, different dilutions of 2C3 were added. The graph shows dose–response of 2C3 on immobilized murine or human PTX3. Human PTX3 was not detected by 2C3 antibody. Evaluation of the amplification efficiency of real‐time RT–PCR assay designed for PTX3 expression in astrocyte cell cultures. (B, C) Melting curve and amplification plot of PTX3 RT–qPCR assay. (D) Standard curves of PTX3 and GAPDH, used as reference mRNA, obtained using fivefold serial dilutions of the cDNA (420, 84, 16.8, 3.36 ng). The threshold cycle ( C t ) values ( y ‐axis) are plotted against log 10 values of cDNA input amounts ( x ‐axis). The graphs are parallel lines and the calculated efficiencies (E) are, respectively, of 1.13 and 1.12 from a y‐slope of −3.04 and −3.07 and a correlation coefficient ( R 2 ) > 0.9.

    Article Snippet: Briefly, 96‐well ELISA plates (Nunc MaxiSorp, Thermo Fischer Scientific, Roskilde, Denmark) were coated with monoclonal antibody 2C3 anti‐mouse PTX3 in coating buffer (15 mM carbonate buffer pH 9.6) and incubated overnight at 4°C.

    Techniques: Enzyme-linked Immunosorbent Assay, Reverse Transcription Polymerase Chain Reaction, Purification, Recombinant, Amplification, Quantitative RT-PCR, Expressing

    Specificity of ELISA and RT–PCR assays A The specificity of the PTX3 ELISA was tested using different dilutions of 2C3 antibody to measure immobilized murine and human PTX3. Purified recombinant murine and human PTX3 were immobilized in 96‐well ELISA plates, and then, different dilutions of 2C3 were added. The graph shows dose–response of 2C3 on immobilized murine or human PTX3. Human PTX3 was not detected by 2C3 antibody. B–D Evaluation of the amplification efficiency of real‐time RT–PCR assay designed for PTX3 expression in astrocyte cell cultures. (B, C) Melting curve and amplification plot of PTX3 RT–qPCR assay. (D) Standard curves of PTX3 and GAPDH, used as reference mRNA, obtained using fivefold serial dilutions of the cDNA (420, 84, 16.8, 3.36 ng). The threshold cycle ( C t ) values ( y ‐axis) are plotted against log 10 values of cDNA input amounts ( x ‐axis). The graphs are parallel lines and the calculated efficiencies (E) are, respectively, of 1.13 and 1.12 from a y‐slope of −3.04 and −3.07 and a correlation coefficient ( R 2 ) > 0.9.

    Journal: The EMBO Journal

    Article Title: Pentraxin 3 regulates synaptic function by inducing AMPA receptor clustering via ECM remodeling and β1‐integrin

    doi: 10.15252/embj.201899529

    Figure Lengend Snippet: Specificity of ELISA and RT–PCR assays A The specificity of the PTX3 ELISA was tested using different dilutions of 2C3 antibody to measure immobilized murine and human PTX3. Purified recombinant murine and human PTX3 were immobilized in 96‐well ELISA plates, and then, different dilutions of 2C3 were added. The graph shows dose–response of 2C3 on immobilized murine or human PTX3. Human PTX3 was not detected by 2C3 antibody. B–D Evaluation of the amplification efficiency of real‐time RT–PCR assay designed for PTX3 expression in astrocyte cell cultures. (B, C) Melting curve and amplification plot of PTX3 RT–qPCR assay. (D) Standard curves of PTX3 and GAPDH, used as reference mRNA, obtained using fivefold serial dilutions of the cDNA (420, 84, 16.8, 3.36 ng). The threshold cycle ( C t ) values ( y ‐axis) are plotted against log 10 values of cDNA input amounts ( x ‐axis). The graphs are parallel lines and the calculated efficiencies (E) are, respectively, of 1.13 and 1.12 from a y‐slope of −3.04 and −3.07 and a correlation coefficient ( R 2 ) > 0.9.

    Article Snippet: Briefly, 96‐well ELISA plates (Nunc MaxiSorp, Thermo Fischer Scientific, Roskilde, Denmark) were coated with monoclonal antibody 2C3 anti‐mouse PTX3 in coating buffer (15 mM carbonate buffer pH 9.6) and incubated overnight at 4°C.

    Techniques: Enzyme-linked Immunosorbent Assay, Reverse Transcription Polymerase Chain Reaction, Purification, Recombinant, Amplification, Quantitative RT-PCR, Expressing

    Description and characterization of the chimeric human FasL-derived constructs Panel A: Schematic representation of soluble FasL (sFasL), Flag-tagged sFasL (sfFasL), polymeric Flag-tagged soluble FasL (pfFasL), polymeric TCR γ4 and δ5 Flag-tagged soluble FasL generating the TCR-pfFasL upon cotransfection, and beta2-microglobulin-fused HLA-A*02: 01 Flag-tagged soluble FasL (HLA-pfFasL). The f and p symbols represent the flag epitope and the LIF receptor-derived domain triggering the polymerisation of the FasL oligomers, respectively. Panel B: direct immunoblot of the supernatants from COS cells transfected with the empty vector (control) or the FasL constructs sFasL, sfFasL and pfFasL. Panel C: immunoprecipitation of the TCR-pfFasL chimera from transfected HEK cells, using an irrelevant IgG1 antibody, the anti-Flag (clone M2), the anti-FasL (clone 10F2), the anti-TCRγδ (clone IMU-510) or the anti-TCRδ5 (clone 12C7) antibodies. Panel D: immunoprecipitation of the HLA-pfFasL chimera from the supernatant of COS cells, with anti-Flag, anti-FasL or anti-β2microglobulin antibodies. As controls, the same experiment was performed with irrelevant IgG1 and IgG2 antibodies. Panel E: cytotoxic effect of the FasL chimeras. The indicated chimeras, as supernatants from transfected cells and quantitated using the ELISA for FasL, were incubated at the indicated concentrations with Jurkat cells. After 18 h, the MTT cell viability assay was performed. The anti-Flag M2 antibody at 0.5 µg/ml was added to sfFasL to render it cytotoxic.

    Journal: PLoS ONE

    Article Title: Enhancing Production and Cytotoxic Activity of Polymeric Soluble FasL-Based Chimeric Proteins by Concomitant Expression of Soluble FasL

    doi: 10.1371/journal.pone.0073375

    Figure Lengend Snippet: Description and characterization of the chimeric human FasL-derived constructs Panel A: Schematic representation of soluble FasL (sFasL), Flag-tagged sFasL (sfFasL), polymeric Flag-tagged soluble FasL (pfFasL), polymeric TCR γ4 and δ5 Flag-tagged soluble FasL generating the TCR-pfFasL upon cotransfection, and beta2-microglobulin-fused HLA-A*02: 01 Flag-tagged soluble FasL (HLA-pfFasL). The f and p symbols represent the flag epitope and the LIF receptor-derived domain triggering the polymerisation of the FasL oligomers, respectively. Panel B: direct immunoblot of the supernatants from COS cells transfected with the empty vector (control) or the FasL constructs sFasL, sfFasL and pfFasL. Panel C: immunoprecipitation of the TCR-pfFasL chimera from transfected HEK cells, using an irrelevant IgG1 antibody, the anti-Flag (clone M2), the anti-FasL (clone 10F2), the anti-TCRγδ (clone IMU-510) or the anti-TCRδ5 (clone 12C7) antibodies. Panel D: immunoprecipitation of the HLA-pfFasL chimera from the supernatant of COS cells, with anti-Flag, anti-FasL or anti-β2microglobulin antibodies. As controls, the same experiment was performed with irrelevant IgG1 and IgG2 antibodies. Panel E: cytotoxic effect of the FasL chimeras. The indicated chimeras, as supernatants from transfected cells and quantitated using the ELISA for FasL, were incubated at the indicated concentrations with Jurkat cells. After 18 h, the MTT cell viability assay was performed. The anti-Flag M2 antibody at 0.5 µg/ml was added to sfFasL to render it cytotoxic.

    Article Snippet: The anti-FasL 14C2 or the anti-Flag mAbs were pre-coated overnight onto 96 well ELISA plates (Maxisorp Nunc, Thermo Scientific, Rochester, USA) respectively at 1 µg or 0.25 µg/well in hydrogenocarbonate coating buffer (pH = 9.6).

    Techniques: Derivative Assay, Construct, Cotransfection, FLAG-tag, Transfection, Plasmid Preparation, Immunoprecipitation, Enzyme-linked Immunosorbent Assay, Incubation, MTT Assay, Viability Assay

    Direct association of sFasL to the pfFasL-containing chimeric proteins during co-expression. Panel A: Identical amounts of pfFasL (1 µg, according to the Flag ELISA) produced in the presence of the indicated ratios of added sFasL plasmid (left panels) was immunoprecipitated with the anti-FasL (upper panel) or anti-Flag (lower panel) antibodies, followed by a SDS-PAGE under reducing conditions and immunoblotting with an anti-FasL antibody. As a control, the same experiment was performed for the sFasL molecule (3 µg according to the FasL ELISA, right panel). Panel B: Densitometric detection and quantification of the pfFasL (grey bars) and the sFasL (black bars) fractions, following transfection of the pfFasL plasmid in the presence of the indicated proportion of the sFasL plasmid. The measures were normalized to the condition lacking sFasL. Mean+/- sd from three experiments. Panel C: The TCR-pfFasL chimera (2 µg, according to an ELISA specific for the TCR-pFasL molecule using anti-TCRδ5 (clone 12C7) and anti-FasL (clone 10F2) as capture and tracing antibodies, respectively), produced in the absence or the presence of the sFasL plasmid at the indicated ratio, was immunoprecipitated with the anti-TCRδ5 antibody, then separated by 10% SDS-PAGE under reducing conditions and revealed by immunoblotting with the anti-FasL antibody. As a control, the immunoprecipitation experiment was performed with 2 µg of sFasL protein. Panel D: COS supernatants containing pfFasL (4 µg/ml according to the Flag ELISA) produced alone, was mixed with culture medium or sFasL (15 µg/ml) produced separately in a total volume of 1 ml, and incubated for 24 h at 37°C. Then the recombinant proteins were immunoprecipitated (left panels) with the anti-FasL (upper panel) or anti-Flag (lower panel) antibodies, followed by a SDS-PAGE under reducing conditions and immunoblotting with an anti-FasL antibody. As a control, the same experiment was performed for the sFasL molecule (15 µg according to the FasL ELISA, right panel).

    Journal: PLoS ONE

    Article Title: Enhancing Production and Cytotoxic Activity of Polymeric Soluble FasL-Based Chimeric Proteins by Concomitant Expression of Soluble FasL

    doi: 10.1371/journal.pone.0073375

    Figure Lengend Snippet: Direct association of sFasL to the pfFasL-containing chimeric proteins during co-expression. Panel A: Identical amounts of pfFasL (1 µg, according to the Flag ELISA) produced in the presence of the indicated ratios of added sFasL plasmid (left panels) was immunoprecipitated with the anti-FasL (upper panel) or anti-Flag (lower panel) antibodies, followed by a SDS-PAGE under reducing conditions and immunoblotting with an anti-FasL antibody. As a control, the same experiment was performed for the sFasL molecule (3 µg according to the FasL ELISA, right panel). Panel B: Densitometric detection and quantification of the pfFasL (grey bars) and the sFasL (black bars) fractions, following transfection of the pfFasL plasmid in the presence of the indicated proportion of the sFasL plasmid. The measures were normalized to the condition lacking sFasL. Mean+/- sd from three experiments. Panel C: The TCR-pfFasL chimera (2 µg, according to an ELISA specific for the TCR-pFasL molecule using anti-TCRδ5 (clone 12C7) and anti-FasL (clone 10F2) as capture and tracing antibodies, respectively), produced in the absence or the presence of the sFasL plasmid at the indicated ratio, was immunoprecipitated with the anti-TCRδ5 antibody, then separated by 10% SDS-PAGE under reducing conditions and revealed by immunoblotting with the anti-FasL antibody. As a control, the immunoprecipitation experiment was performed with 2 µg of sFasL protein. Panel D: COS supernatants containing pfFasL (4 µg/ml according to the Flag ELISA) produced alone, was mixed with culture medium or sFasL (15 µg/ml) produced separately in a total volume of 1 ml, and incubated for 24 h at 37°C. Then the recombinant proteins were immunoprecipitated (left panels) with the anti-FasL (upper panel) or anti-Flag (lower panel) antibodies, followed by a SDS-PAGE under reducing conditions and immunoblotting with an anti-FasL antibody. As a control, the same experiment was performed for the sFasL molecule (15 µg according to the FasL ELISA, right panel).

    Article Snippet: The anti-FasL 14C2 or the anti-Flag mAbs were pre-coated overnight onto 96 well ELISA plates (Maxisorp Nunc, Thermo Scientific, Rochester, USA) respectively at 1 µg or 0.25 µg/well in hydrogenocarbonate coating buffer (pH = 9.6).

    Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Produced, Plasmid Preparation, Immunoprecipitation, SDS Page, Transfection, Incubation, Recombinant

    Effect of sFasL on the supernatant production of the Flag-tagged FasL constructs. Panels A to D : An increasing amount expressed in percentage, of the sFasL encoding plasmid, was co-transfected with a fixed amount of the plasmids encoding sfFasL (Panel A), pfFasL (Panel B), TCR-pfFasL (Panel C) and HLA-pfFasL (Panel D). The secreted proteins were quantified in culture supernatants using an ELISA specific for FasL (shaded histograms, right-hand scale) and for Flag-tagged FasL (curves, left-hand scale). For the Flag ELISA, the measured concentrations were normalized according to the condition lacking sFasL. Are presented the mean +/- sd of four independent transfection experiments. * 0.02≤p≤0.05; ** p≤0.02. Panel E : direct anti-FasL immunoblot analysis of identical volumes of the cell culture supernatant containing pfFasL produced alone and with 50% of the sFasL plasmid, after SDS-PAGE separation under reducing conditions.

    Journal: PLoS ONE

    Article Title: Enhancing Production and Cytotoxic Activity of Polymeric Soluble FasL-Based Chimeric Proteins by Concomitant Expression of Soluble FasL

    doi: 10.1371/journal.pone.0073375

    Figure Lengend Snippet: Effect of sFasL on the supernatant production of the Flag-tagged FasL constructs. Panels A to D : An increasing amount expressed in percentage, of the sFasL encoding plasmid, was co-transfected with a fixed amount of the plasmids encoding sfFasL (Panel A), pfFasL (Panel B), TCR-pfFasL (Panel C) and HLA-pfFasL (Panel D). The secreted proteins were quantified in culture supernatants using an ELISA specific for FasL (shaded histograms, right-hand scale) and for Flag-tagged FasL (curves, left-hand scale). For the Flag ELISA, the measured concentrations were normalized according to the condition lacking sFasL. Are presented the mean +/- sd of four independent transfection experiments. * 0.02≤p≤0.05; ** p≤0.02. Panel E : direct anti-FasL immunoblot analysis of identical volumes of the cell culture supernatant containing pfFasL produced alone and with 50% of the sFasL plasmid, after SDS-PAGE separation under reducing conditions.

    Article Snippet: The anti-FasL 14C2 or the anti-Flag mAbs were pre-coated overnight onto 96 well ELISA plates (Maxisorp Nunc, Thermo Scientific, Rochester, USA) respectively at 1 µg or 0.25 µg/well in hydrogenocarbonate coating buffer (pH = 9.6).

    Techniques: Construct, Plasmid Preparation, Transfection, Enzyme-linked Immunosorbent Assay, Cell Culture, Produced, SDS Page

    Effect of sFasL on cell targeting of the FasL-containing chimeras. Panel A : Schematic description of the experimental model used. The chimera is enriched at the surface of the CD32-expressing L-cells via its HLA targeting module and an anti-HLA monoclonal antibody. Panel B: murine Fas (continuous line), human CD32 (dashed line) and IgG1 isotype-matched control (shaded histogram) staining of the CD32+ L-cell transfectant. Living cells were gated on the basis of the morphological parameters. Panel C : Fas sensitivity of the CD32+ L-cell transfectant to the indicated concentrations of the anti-Fas JO-2 antibody (circles), the HLA-pfFasL chimera expressed alone (triangle) or in the presence of 25% of the sFasL plasmid (squares), in the MTT viability assay. Panel D : The CD32+ L-cells were incubated with the HLA-pfFasL chimera produced in the presence (black bars) or in the absence (white bars) of 25% of the sFasL plasmid, together with the indicated irrelevant IgG1 isotype-matched, anti-beta-2 microglobulin or anti-Flag antibodies. The concentrations of the chimera that triggered 15% of cell death and were at 15 and 0.3 ng/ml in the absence and presence of sFasL, as estimated using the ELISA specific for the Flag-tagged FasL. Cytotoxicity was measured with the propidium iodide assay and normalized to the effect of the chimera in the absence of antibody. Are presented the mean +/- sd of three independent experiments. Panel E: reversal in the presence of the blocking anti-FasL and anti-CD32 antibodies, of the cytotoxic effect of the immune complexes between the anti-Flag antibody and HLA-pfFasL co-expressed with sFasL. Are presented the mean +/- sd of three independent experiments. ns : non significant ; ** p≤0.02.

    Journal: PLoS ONE

    Article Title: Enhancing Production and Cytotoxic Activity of Polymeric Soluble FasL-Based Chimeric Proteins by Concomitant Expression of Soluble FasL

    doi: 10.1371/journal.pone.0073375

    Figure Lengend Snippet: Effect of sFasL on cell targeting of the FasL-containing chimeras. Panel A : Schematic description of the experimental model used. The chimera is enriched at the surface of the CD32-expressing L-cells via its HLA targeting module and an anti-HLA monoclonal antibody. Panel B: murine Fas (continuous line), human CD32 (dashed line) and IgG1 isotype-matched control (shaded histogram) staining of the CD32+ L-cell transfectant. Living cells were gated on the basis of the morphological parameters. Panel C : Fas sensitivity of the CD32+ L-cell transfectant to the indicated concentrations of the anti-Fas JO-2 antibody (circles), the HLA-pfFasL chimera expressed alone (triangle) or in the presence of 25% of the sFasL plasmid (squares), in the MTT viability assay. Panel D : The CD32+ L-cells were incubated with the HLA-pfFasL chimera produced in the presence (black bars) or in the absence (white bars) of 25% of the sFasL plasmid, together with the indicated irrelevant IgG1 isotype-matched, anti-beta-2 microglobulin or anti-Flag antibodies. The concentrations of the chimera that triggered 15% of cell death and were at 15 and 0.3 ng/ml in the absence and presence of sFasL, as estimated using the ELISA specific for the Flag-tagged FasL. Cytotoxicity was measured with the propidium iodide assay and normalized to the effect of the chimera in the absence of antibody. Are presented the mean +/- sd of three independent experiments. Panel E: reversal in the presence of the blocking anti-FasL and anti-CD32 antibodies, of the cytotoxic effect of the immune complexes between the anti-Flag antibody and HLA-pfFasL co-expressed with sFasL. Are presented the mean +/- sd of three independent experiments. ns : non significant ; ** p≤0.02.

    Article Snippet: The anti-FasL 14C2 or the anti-Flag mAbs were pre-coated overnight onto 96 well ELISA plates (Maxisorp Nunc, Thermo Scientific, Rochester, USA) respectively at 1 µg or 0.25 µg/well in hydrogenocarbonate coating buffer (pH = 9.6).

    Techniques: Expressing, Staining, Transfection, Plasmid Preparation, MTT Assay, Viability Assay, Incubation, Produced, Enzyme-linked Immunosorbent Assay, Blocking Assay

    Effect of sFasL on the cytotoxic activity of the Flag-tagged FasL chimeras. The FasL-derived proteins sfFasL (Panel A), pfFasL (Panel B), TCR-pfFasL (Panel C) and HLA-pfFasL (Panel D) were expressed alone or upon co-transfection with the indicated percentage of the plasmid encoding sFasL. A fixed concentration triggering 25 to 40% of cell death (1.9 ng/ml for sfFasL, 0.6 ng/ml for pfFasL, 0.7 ng/ml for HLA-pfFasL and 2.2 ng/ml for TCR-pfFasL), for the FasL-derived protein quantitated with the ELISA specific for Flag-tagged FasL, was incubated with the Fas-sensitive Jurkat cells. For the sfFasL construct, the filled squares and the empty squares depict the cytotoxicity of sfFasL in the presence and absence of the cross-linking anti-Flag antibody at 0.5 µg/ml), respectively. Cytotoxicity was estimated by a measure of the remaining viable cells using the MTT assay. Are presented the mean +/- sd of four independent transfection experiments. * 0.01≤p≤0.05; ** p≤0.01.

    Journal: PLoS ONE

    Article Title: Enhancing Production and Cytotoxic Activity of Polymeric Soluble FasL-Based Chimeric Proteins by Concomitant Expression of Soluble FasL

    doi: 10.1371/journal.pone.0073375

    Figure Lengend Snippet: Effect of sFasL on the cytotoxic activity of the Flag-tagged FasL chimeras. The FasL-derived proteins sfFasL (Panel A), pfFasL (Panel B), TCR-pfFasL (Panel C) and HLA-pfFasL (Panel D) were expressed alone or upon co-transfection with the indicated percentage of the plasmid encoding sFasL. A fixed concentration triggering 25 to 40% of cell death (1.9 ng/ml for sfFasL, 0.6 ng/ml for pfFasL, 0.7 ng/ml for HLA-pfFasL and 2.2 ng/ml for TCR-pfFasL), for the FasL-derived protein quantitated with the ELISA specific for Flag-tagged FasL, was incubated with the Fas-sensitive Jurkat cells. For the sfFasL construct, the filled squares and the empty squares depict the cytotoxicity of sfFasL in the presence and absence of the cross-linking anti-Flag antibody at 0.5 µg/ml), respectively. Cytotoxicity was estimated by a measure of the remaining viable cells using the MTT assay. Are presented the mean +/- sd of four independent transfection experiments. * 0.01≤p≤0.05; ** p≤0.01.

    Article Snippet: The anti-FasL 14C2 or the anti-Flag mAbs were pre-coated overnight onto 96 well ELISA plates (Maxisorp Nunc, Thermo Scientific, Rochester, USA) respectively at 1 µg or 0.25 µg/well in hydrogenocarbonate coating buffer (pH = 9.6).

    Techniques: Activity Assay, Derivative Assay, Cotransfection, Plasmid Preparation, Concentration Assay, Enzyme-linked Immunosorbent Assay, Incubation, Construct, MTT Assay, Transfection

    Analysis of GRP78–HTJ1 and OxPL–GRP78 interactions. (A) GRP78 and HTJ1 expression in HPAEC and human lung microvascular endothelial cells was detected by Western Blot. (B, C) GRP78 interactions were analyzed in coimmunoprecipitation assays using lysates from control or DMPC- (10 μg/ml, 15 min) or OxPAPC-stimulated (10 μg/ml) cells with antibody to GRP78 (B, top), HTJ1 (B, bottom), or EO6 antibody recognizing OxPL (C). (D) Human recombinant GRP78 was incubated with OxPAPC, OxPAPS, or their oxidation-resistant analogues DMPC or DMPS. Left, native gel electrophoresis, followed by Western blot with anti-GRP78 antibody. Shift in electrophoretic mobility of GRP78 incubated with OxPAPS, but not DMPS, indicates formation of GRP78–OxPAPS complex. Right, SDS–PAGE, followed by Western blot with EO6 antibody and reprobing with anti-GRP78 antibody. Positive EO6 immunoreactivity of GRP78 preincubated with OxPAPC indicates formation of GRP78–OxPAPC complex. (E) ELISA plates coated with OxPAPS or DMPS or control uncoated plates incubated with PBS incubated with human recombinant GRP78 (left) or HPAEC lysates (middle and right). The bound GRP78 was detected using anti-GRP78 antibody. * p

    Journal: Molecular Biology of the Cell

    Article Title: GRP78 is a novel receptor initiating a vascular barrier protective response to oxidized phospholipids

    doi: 10.1091/mbc.E13-12-0743

    Figure Lengend Snippet: Analysis of GRP78–HTJ1 and OxPL–GRP78 interactions. (A) GRP78 and HTJ1 expression in HPAEC and human lung microvascular endothelial cells was detected by Western Blot. (B, C) GRP78 interactions were analyzed in coimmunoprecipitation assays using lysates from control or DMPC- (10 μg/ml, 15 min) or OxPAPC-stimulated (10 μg/ml) cells with antibody to GRP78 (B, top), HTJ1 (B, bottom), or EO6 antibody recognizing OxPL (C). (D) Human recombinant GRP78 was incubated with OxPAPC, OxPAPS, or their oxidation-resistant analogues DMPC or DMPS. Left, native gel electrophoresis, followed by Western blot with anti-GRP78 antibody. Shift in electrophoretic mobility of GRP78 incubated with OxPAPS, but not DMPS, indicates formation of GRP78–OxPAPS complex. Right, SDS–PAGE, followed by Western blot with EO6 antibody and reprobing with anti-GRP78 antibody. Positive EO6 immunoreactivity of GRP78 preincubated with OxPAPC indicates formation of GRP78–OxPAPC complex. (E) ELISA plates coated with OxPAPS or DMPS or control uncoated plates incubated with PBS incubated with human recombinant GRP78 (left) or HPAEC lysates (middle and right). The bound GRP78 was detected using anti-GRP78 antibody. * p

    Article Snippet: GRP78 ELISA Microtiter 96-well plates (MaxiSorp; Nunc, Thermo Scientific, Rochester, NY) were coated with OxPAPS or DMPS (each 100 μg/ml in PBS containing 0.01% BHT) at 4°C overnight.

    Techniques: Expressing, Western Blot, Recombinant, Incubation, Nucleic Acid Electrophoresis, SDS Page, Enzyme-linked Immunosorbent Assay