Tumor lysates were created using normal colon and colon cancer cell line LS174T (ATCC, Manassas, VA) xenografts. Human normal colon was obtained under standard sterile conditions in the operating room at UCSD Thornton hospital under UCSD IRB protocol 140046 with informed patient consent. Western blotting procedures were carried out at room temperature. Protein samples were isolated and transferred to Trans Blot turbo Mini-size nitrocellulose membranes (Bio-Rad, Cat. 1704270) using the Bio-Rad Trans-Blot Turbo Transfer System. The membrane was then incubated with antibody conjugated with IRDye800CW at 4°C overnight. The membrane was scanned with a LI-COR Odyssey Infrared Imaging System model 9120 (LI-COR, Lincoln, NE) and detection and quantification of band intensities was conducted using Image Studio Lite Version 5.2 software (LI-COR, Lincoln, NE). Bands were normalized to total protein by dividing the intensity of the band by the intensity of the total protein from the same sample on the same blot. β-actin was utilized as a standard.
Trans blot turbo mini size nitrocellulose membranes
Manufactured by Bio-Rad
The Trans Blot Turbo Mini-size nitrocellulose membranes are a type of laboratory equipment used for the transfer and immobilization of proteins from electrophoretic gels to a membrane for further analysis. The membranes provide a stable and consistent surface for the binding of proteins, allowing for various downstream applications such as immunodetection.
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Lab products found in correlation
Odyssey infrared imaging system model 9120, by LI COR (1 mentions)
Image studio lite version 5, by LI COR (1 mentions)
Ls174t, by American Type Culture Collection (1 mentions)
Trans blot turbo transfer system, by Bio-Rad (1 mentions)
Rat anti ha clone 3f10, by Roche (1 mentions)
Irdye 800cw donkey anti rabbit igg secondary antibody, by LI COR (1 mentions)
Odyssey infrared imaging system, by LI COR (1 mentions)
2 protocols using trans blot turbo mini size nitrocellulose membranes
1
Western Blot Analysis of Colon Cancer
2
Western Blot Analysis of Parasite Proteins
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Percoll-enriched schizonts were resuspended in PBS, solubilized in protein loading buffer, and denatured at 95°C for 10 min. Parasite extracts were resolved by SDS-PAGE gel electrophoresis, then transferred to Transblot turbo mini-size nitrocellulose membranes (Bio-Rad) and blocked for 1 hr in 3% BSA in PBS at 4°C. For detecting FIKK4.1-HA, membranes were probed with rat anti-HA (clone 3F10, Roche; 1:1000) followed by 1:20,000 IRDye 680RD goat anti-rat IgG secondary antibody (LI-COR Biosciences). For detecting KAHRP and PTP4-V5, membranes were probed with rabbit anti-V5 (MA5-32053, Invitrogen; 1:200), followed by 1:20,000 IRDye 800CW donkey anti-rabbit IgG secondary antibody (LI-COR Biosciences). For detecting biotinylated proteins, blots were probed with fluorophore-conjugated streptavidin IRDye 800CW (LI-COR Biosciences) at a 1:1000 dilution. All antibody incubations were performed in 3% BSA in PBS for 1–2 hr. Blots were washed 3× in PBS with 0.2% Tween 20. Blots were visualized using the Odyssey infrared imaging system (LI-COR Biosciences).
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