For YebF samples induced 16–20 h, cells were pelleted, and the supernatant was harvested and precipitated with ice cold 10% trichloroacetic acid (TCA). For 4 h induction samples, culture volumes containing both cells and supernatant were harvested and directly TCA precipitated. For scFv13-R4, periplasmic fractions were harvested after spheroplasting cells in buffers containing 0.2 M Tris-Ac (pH 8.2), 0.25 M sucrose, 160 μg/ml lysozyme, and 0.25 mM EDTA. For RNaseA, protein was purified by nickel-affinity chromatography from the periplasmic fractions of 50–100 ml cultures. In all cases, protein was solubilized in Laemmli sample buffer and resolved on SDS-polyacrylamide gels (BioRad). Western blotting used 6x-His tag-specific polyclonal antibodies (Abcam) or C. jejuni heptasaccharide glycan-specific antiserum hR6 29 (link). Pierce enhanced chemiluminescent (ECL) substrate (Thermo Scientific) was used for detection of bound antibodies. All blots were visualized using a Chemidoc™ XRS+ system with Image Lab™ image capture software (BioRad).
Image lab image capture software
Manufactured by Bio-Rad
Image Lab™ image capture software is a versatile tool designed for image acquisition, analysis, and documentation. It provides a user-friendly interface for capturing and managing images from a variety of imaging devices, including gel documentation systems, western blot imagers, and other compatible hardware.
Automatically generated - may contain errors
Lab products found in correlation
Chemidoc xrs system, by Bio-Rad (2 mentions)
Sds polyacrylamide gel, by Bio-Rad (1 mentions)
Pierce enhanced chemiluminescent ecl substrate, by Thermo Fisher Scientific (1 mentions)
His tag specific polyclonal antibodies, by Abcam (1 mentions)
Trypsin edta, by Thermo Fisher Scientific (1 mentions)
Hrp conjugated goat anti mouse igg, by Thermo Fisher Scientific (1 mentions)
Tisw28 rotor, by Beckman Coulter (1 mentions)
2 protocols using image lab image capture software
1
Protein Purification and Analysis Protocol
2
Quantitative Western Blot Analysis of Pseudotyped Viral Particles
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Pseudotyped particles were concentrated using an ultracentrifuge at 20,000 rpm in a TiSW28 rotor (Beckman-Coulter) for 2 h at 4 °C. The pellets were resuspended in 30 μL of 3 μg/mL trypsin EDTA (Thermo Fisher Scientific) solution, and placed in a 37 °C water bath for 15 min. The samples were then analyzed by Western blot using goat anti-A/HongKong/1/68 (H3) (NR-3118, BEI resources), goat anti-A/shorebird/Delaware/127/1997(N2) (NR-670, BEI resources) and mouse anti-VSV-M antibodies (Kerafast) followed by incubation with HRP-conjugated rabbit anti-goat IgG (Life Technologies) and HRP-conjugated goat anti-mouse IgG (Life Technologies). All Western blots were visualized and analyzed using a Chemidoc XRS+ system with Image Lab image capture software (BioRad). TheImage Lab software was used to quantify intensities of bands using low sensitivity setting. The software detects the bands by the signal contrast between band and background. We note that all bands have only been adjusted in contrast and brightness in the Image Lab software. The software shows the original signal intensities of the bands.
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