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β-casein

Manufactured by Abbiotec
Sourced in United States

β-casein is a milk protein that serves as a major component of the casein micelle structure in bovine milk. It plays a crucial role in the formation and stability of milk emulsions and gels.

Automatically generated - may contain errors

3 protocols using β-casein

1

Immunofluorescence Characterization of Bovine Mammary Epithelial Cells

Immunofluorescence assay was carried out as described previously [29 (link)]. BMEC was probed with primary antibodies against the following: β-casein (1:50, Abbiotec, 251309), cytokeratin 18 (Santa Cruz, CA, USA, sc-51582), Tudor-SN (Abcam, ab 71186), p-Stat5 (Tyr694) (Cell Signaling Technology, #9359). Alexa Fluor 488 or Alexa Fluor 647 secondary antibodies (ZSGB-BIO, Beijing, China) were used to detect primary antibodies. Following PI or DAPI staining, the cells were imaged under a TCS-SP2 AOBS confocal microscope (Leica, Heidelberg, Germany).
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Western blotting analysis was performed using standard techniques reported by Huang [55 (link)]. Total cell lysate containing about protein 30 μg was separated on a 10% SDS-PAGE gel and transferred onto nitrocellulose membranes (Bio-RAD Company, Hercules, CA, USA). Membranes were blocked in 5% skim milk (in Tris-buffered saline with 5% skim milk and 0.1% Tween-20). Membranes were probed with primary antibodies specific for the following antibodies: LeuRS, GLUT1, Cyclin D1, SREBP-1c (the mature form, 68 kD) (Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA); p-mTOR, mTOR, p-S6K1, S6K1, (Cell Signaling Technology, Beverly, MA, USA); β-Casein (Abbiotec, San Diego, CA, USA) and β-Actin (Santa Cruz, CA, USA), followed by a second incubation with secondary antibodies (1:1000) conjugated to HRP (ZSGB-BIO, Beijing, China). The chemiluminescence detection of HRP-conjugated secondary antibodies was performed using Super ECL plus (ApplyGEN, Beijing, China). Analysis of these western blotting results were subsequently performed using the Quantity One software.
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Western blot analysis was performed as described previously [23 (link),29 (link)]. Briefly, 30 μg of protein equivalents of whole-cell extracts was separated on 10% SDS-PAGE. Gels were transferred to nitrocellulose membranes, then the membranes were blocked in 5% skim milk in pH 7.5 Tris-buffered saline with 0.1% Tween (TBST) for 2 h at 37 °C, then incubated with primary antibodies in 5% skim milk/TBST overnight at 4 °C, and then incubated with horseradish peroxidase-conjugated IgG in 5% skim milk/TBST for 1.5 h at 37 °C. The chemiluminescence detection of HRP-conjugated secondary antibodies was performed using Super ECL plus (ApplyGEN, Beijing, China).
Primary antibodies against the following proteins were used: Tudor-SN (ab 71186), p-NFκB1 p105/p50 (S337) (ab28849), Abcam, Cambridge, MA, USA; mTOR (#2983), p-mTOR (Ser2448) (#5536), and p-Stat5 (Tyr694) (#9359), Cell Signaling Technology, Beverly, MA, USA; cyclinD1 (sc-753), Stat5 (sc-28685), SREBP-1c (the mature form, 68 kD) (sc-365513), β-actin (sc-47778), Santa Cruz, CA, USA; β-casein (Abbiotec, San Diego, CA, USA, 251309).
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