Cell lysates were prepared from 106 induced cells in 15 µl RIPA buffer (150 mM NaCl, 50 mM Tris, pH 8, 1% NP-40, and 0.1% SDS) and kept on ice for 10 min, followed by incubation with 1 µl of benzonase (EMD Millipore) for 20 min at 37°C. Extracts were separated by 10% SDS-PAGE and transferred to nitrocellulose membranes. After a 30-min block in TBS-T/5% milk (TBS, 0.1% Tween 20, and 5% powder nonfat milk), membranes were incubated overnight at 4°C with anti-CAL1 (rabbit, 1:1,000; Erhardt et al., 2008 (link)), anti–CENP-A (rabbit, 1:1,000; a gift from G. Karpen, Lawrence Berkeley National Laboratory and University of California, Berkeley, Berkeley, CA), anti–CENP-C (guinea pig, 1:3,000), and anti-LacI (mouse, 1:100; EMD Millipore). Anti–α-tubulin (mouse, 1:3,000; Sigma-Aldrich) was used as a loading control.
Anti cenp a
Manufactured by Merck Group
Anti–CENP-A is a laboratory product that detects the presence of the CENP-A protein in samples. CENP-A is a key component of the centromere, which is the central region of a chromosome. This product can be used for research purposes related to the study of the centromere and chromosome structure.
Automatically generated - may contain errors
Lab products found in correlation
Benzonase, by Merck Group (1 mentions)
Anti α tubulin, by Merck Group (1 mentions)
Anti laci, by Merck Group (1 mentions)
Goat anti mouse igg secondary antibody, by LI COR (1 mentions)
Anti actin, by Thermo Fisher Scientific (1 mentions)
Anti h3k4me2, by Merck Group (1 mentions)
Anti smc1, by Fortis Life Sciences (1 mentions)
Goat anti rabbit igg secondary antibody, by LI COR (1 mentions)
Anti myc, by Roche (1 mentions)
Anti h3k9me3, by Abcam (1 mentions)
2 protocols using anti cenp a
1
Immunoblot Analysis of Centromeric Proteins
2
Antibody Characterization Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
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The following antibodies were used in this study: anti-centromere antibody (ACA; or CREST-ImmunoVision; HCT-0100), anti-Myc (Roche; 11667203001), anti-Smc1 (Bethyl; A300-055A), anti-Rpb1 (Abcam; ab5408), anti-H3K4me2 (EMD Millipore; 07–030), anti-H3K9me3 (Abcam; ab8898), anti-Actin (Invitrogen; MA5-11869), anti-CENP-B (Abcam; ab25734), anti–CENP-A (EMD Millipore; 07–574), and anti–Rpb2-pSer2 (BioLegend; H5). Anti-Sgo1, anti-Bub1, and anti-Wapl were made in-house as described previously (Liu et al., 2013a (link); Qu et al., 2019 (link)). Anti-Sororin antibodies described previously were a gift from Dr. Susannah Rankin at Oklahoma Medical Research Foundation, Oklahoma City, OK (Liu et al., 2013b (link)).
The secondary antibodies were purchased from LI-COR: Goat anti-Mouse IgG Secondary Antibody (926–68070) and Goat anti-Rabbit IgG Secondary Antibody (926–32211).
For immunoblotting, primary and secondary antibodies were used at 1-µg ml−1 concentration.
The secondary antibodies were purchased from LI-COR: Goat anti-Mouse IgG Secondary Antibody (926–68070) and Goat anti-Rabbit IgG Secondary Antibody (926–32211).
For immunoblotting, primary and secondary antibodies were used at 1-µg ml−1 concentration.
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