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Sds containing sample buffer

Manufactured by Bio-Rad

SDS-containing sample buffer is a solution used in protein electrophoresis to prepare samples for analysis. It contains the anionic detergent sodium dodecyl sulfate (SDS), which denatures proteins and imparts a uniform negative charge, allowing for separation based on molecular weight.

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2 protocols using sds containing sample buffer

1

Dilution and Preparation of rFVIII

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1 vial of Helixate FS (Bayer Healthcare) containing 252 IU was diluted with 2 ml water (final concentration = 0.126 IU/μl containing 12.6 ng/μl of rFVIII protein). For gel separation, 22 μl of 12.6 ng/μl rFVIII was diluted with 10 μl of SDS-containing sample buffer (BioRad) and heated at 95°C for 5 min. 15 μl of this mixture was loaded per gel lane containing ~130 ng of rFVIII.
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2

Quantification of Tumor-derived scIgGs

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Tumor lysates were prepared in RIPA buffer (ThermoFisher) containing a 10% protease inhibitor cocktail (ThermoFisher) using a gentleMACS device (Miltenyi Biotech) based on the manufacture’s protocol. For ELISA detection, anti-hinge rabbit polyclonal antibodies (2 μg/ml) were coated on high-binding 96-well plates to capture scIgGs in tumor extracts; and anti-mouse specific antibody at 1:5000 dilution (Jackson Immune Research Laboratory, PA) with HRP conjugation was used for detection. For Western blotting detection, protein A magnetic beads were incubated with cell lysates at 4 °C for 1 hour to capture total IgGs including both intact IgGs and scIgGs, and the beads were collected and incubated with SDS containing sample buffer (Bio-Rad) at 95 °C for 5 min. The eluted samples were subjected to SDS-PAGE separation and WB detection using a goat anti-mouse IgG-Fc-HRP conjugate (1:4000) (Jackson Immune Research Laboratory, PA) as previously described10 (link),12 (link).
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