The specific antibody response against H7N9 was determined by ELISA. In brief, 50 μL of 0.5 μg/mL HA protein (H7N9 WV vaccine bulk) in 0.1 M carbonate buffer (pH 9.5) was coated onto 96-well microplates by overnight incubation at 4°C. The coated plates were washed twice with 0.05% Tween 20 in PBS and then blocked with 3% BSA in PBS at room temperature for 1 h. Diluted sera from immunized animals were added to the wells and incubated for 2 h at room temperature. HRP-conjugated goat anti-mouse IgG (1:10000; Thermo Scientific), HRP-conjugated rabbit anti-mouse IgG1 (1:5000; Invitrogen), and HRP-conjugated rabbit anti-mouse IgG2a (1: 5000; Invitrogen) were used as the secondary antibodies. The assay was developed by using the TMB substrate set (BioLegend). The absorbance was measured using a SpectraMax M2 microplate reader (Molecular Device) at 450 nm.
Hrp conjugated rabbit anti mouse igg1
Manufactured by Thermo Fisher Scientific
HRP-conjugated rabbit anti-mouse IgG1 is a secondary antibody reagent used for the detection of mouse IgG1 in various immunoassay applications. The antibody is conjugated with horseradish peroxidase (HRP) enzyme, which enables signal amplification and sensitive detection.
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Lab products found in correlation
Tmb substrate set, by BioLegend (1 mentions)
Hrp conjugated rabbit anti mouse igg2a, by Thermo Fisher Scientific (1 mentions)
Spectramax m2 microplate reader, by Molecular Devices (1 mentions)
Hrp conjugated goat anti mouse igg, by Thermo Fisher Scientific (1 mentions)
Enhanced chemiluminescence method, by Cytiva (1 mentions)
Mabcam 8226, by Abcam (1 mentions)
Sds page gel, by Bio-Rad (1 mentions)
2 protocols using hrp conjugated rabbit anti mouse igg1
1
Quantifying H7N9 Antibody Response by ELISA
2
Western Blot Analysis of iNOS
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BM-MDSCs and control spleen cells were lysed in cold RIPA buffer containing a Halt protease inhibitor mixture (Pierce/Thermo Fisher, Rockford, IL), and the protein content was determined using the bichinconic acid assay (Pierce). Proteins from cell lysates (20 µg protein each) were loaded onto and resolved in 7.5% SDS-PAGE gels (Bio-Rad) under reducing conditions. The proteins were then transferred to nitrocellulose membranes. The membranes were blotted with an anti-mouse iNOS mAb (mouse mAb, Cat. No. sc-7271; Santa Cruz Biotechnology, Dallas, TX) at a 1∶500 dilution. Horseradish peroxidase (HRP)-conjugated rabbit anti-mouse IgG1 (Invitrogen) was used as a secondary Ab at a 1∶10,000 dilution. The protein bands were visualized using the enhanced chemiluminescence method (Amersham/GE Healthcare Life Sciences, Piscataway, NJ). The membranes were stripped and re-probed with a HRP-conjugated mAb to β-actin (mouse mAb, clone mAbcam 8226; Abcam, Cambridge, MA) at a 1∶5,000 dilution to ensure equal sample loading.
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