Cells were lysed with radioimmunoprecipitation assay buffer and whole cell lysates were subjected to 10% SDS-PAGE and transferred onto polyvinylidene difluoride membranes (Thermo Fisher Scientific). The membranes were incubated with the anti-Sca-1 mouse monoclonal antibody (1:1,000), anti-c-kit rabbit polyclonal antibody (1:1,000; sc-168; Santa Cruz Biotechnology, Inc.), anti-osterix rabbit polyclonal antibody (1:1,000), anti-RUNX2 (runt-related transcription factor 2) rabbit polyclonal antibody (1:,000; sc-10758; Santa Cruz Biotechnology, Inc.), anti-fibronectin mouse monoclonal antibody (1:1,000; sc-59826; Santa Cruz Biotechnology, Inc.), anti-osteocalcin rabbit polyclonal antibody (1:1,000) and anti-osteopontin rabbit polyclonal antibody (1:1,000; sc-20788; Santa Cruz Biotechnology, Inc.). The membranes were then incubated with Novex® alkaline-phosphatase-conjugated goat anti-rabbit polyclonal secondary antibody (WP20006; Thermo Fisher Scientific), or goat anti-mouse monoclonal secondary antibody (WP20007; Thermo Fisher Scientific) according to manufacturer’s instructions. The antibodies were detected using a chromogenic immunodetection system (WesternBreeze; Thermo Fisher Scientific) according to the manufacturer’s instructions. The anti-α-tubulin monoclonal antibody (Santa Cruz Biotechnology, Inc.) was used for the normalization of western blot analyses.
Sc 20788
Manufactured by Santa Cruz Biotechnology
Sc-20788 is a laboratory equipment product offered by Santa Cruz Biotechnology. It is designed for use in scientific research applications. The core function of this product is to facilitate specific laboratory processes, but a detailed description cannot be provided while maintaining an unbiased and factual approach.
Automatically generated - may contain errors
Lab products found in correlation
Western breeze, by Thermo Fisher Scientific (1 mentions)
Anti α tubulin monoclonal antibody, by Santa Cruz Biotechnology (1 mentions)
Polyvinylidene difluoride membrane, by Thermo Fisher Scientific (1 mentions)
Sc 10758, by Santa Cruz Biotechnology (1 mentions)
Sc 59826, by Santa Cruz Biotechnology (1 mentions)
Anti fibronectin mouse monoclonal antibody, by Santa Cruz Biotechnology (1 mentions)
Sc 168, by Santa Cruz Biotechnology (1 mentions)
Triton x 100, by Merck Group (1 mentions)
Alexa fluor 555, by Thermo Fisher Scientific (1 mentions)
Alexa fluor 647 conjugated secondary antibody, by Thermo Fisher Scientific (1 mentions)
12 well plate, by BD (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions)
Paraformaldehyde (pfa), by Electron Microscopy Sciences (1 mentions)
Mab2627, by Merck Group (1 mentions)
2 protocols using sc 20788
1
Western Blot Analysis of Stem Cell Markers
2
Immunofluorescence Analysis of VCAM-1 and OPN
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To investigate VCAM-1 and OPN expression level of murine ECs, MSCs, and osteoblasts in different conditions, the cells were respectively seeded onto a cover glass in 12-well plates (Falcon) and cocultured with B-ALL cells for 24 hours or infused into devices for 3D culture. Cells were fixed with 4% paraformaldehyde (15711, Electron Microscopy Sciences) for 30 min, permeabilized with 0.3% Triton X-100 (11332481001, Sigma-Aldrich) for 20 min, and then blocked with 3% bovine serum albumin for 1 hour on ice to prevent nonspecific binding. Cells were incubated with VCAM-1 (1:200; MAB2627, Millipore) and OPN (1:50; sc-20788, Santa Cruz Biotechnology) primary antibodies for 3 hours at RT or overnight at 4°C and then visualized with Alexa Fluor 555– or Alexa Fluor 647–conjugated secondary antibodies (Thermo Fisher Scientific). Cells were incubated for 10 min with 4′,6-diamidino-2-phenylindole (DAPI; Thermo Fisher Scientific) to counterstain nuclei.
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