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Anti cenp a

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The Anti-CENP-A is a laboratory reagent used to detect the presence of the CENP-A protein, a key component of the centromere in eukaryotic cells. It serves as a tool for researchers to study chromosome structure and dynamics.

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Lab products found in correlation

Alexa fluor 488 phalloidin, by Thermo Fisher Scientific (1 mentions) Alexa fluor 647 goat anti rabbit igg h l, by Thermo Fisher Scientific (1 mentions) F actin, by Thermo Fisher Scientific (1 mentions) Anti phospho histone h3 ser10, by Merck Group (1 mentions) Anti lamin a, by Merck Group (1 mentions) Anti kif18a, by Fortis Life Sciences (1 mentions) Anti α tubulin dm1a, by Merck Group (1 mentions) Alexa fluor 488 goat anti rabbit, by Thermo Fisher Scientific (1 mentions) Image studio software, by LI COR (1 mentions) Ab185059, by Abcam (1 mentions) Kwikquant imager, by Kindle Biosciences (1 mentions) Ab32072, by Thermo Fisher Scientific (1 mentions) Jbw301, by Abcam (1 mentions)

2 protocols using anti cenp a

1

Immunofluorescence and Western Blotting Protocols

Cited 1 time
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Immunofluorescence and Western blotting were performed essentially as previously described [84 (link)]. For immunofluorescence, the following antibodies were used. Primary antibodies: anti-Lamin A (1:1000; Sigma-Aldrich), anti-centromere protein (ACA, CREST, 1:400; Antibodies Incorporated), anti-phospho-Histone H3 (Ser10, 1:300; Sigma-Aldrich), anti-α-tubulin DM1A (1:500; Sigma-Aldrich), anti-CENP-A (3–19, 1:300; Invitrogen). Secondary antibodies: Alexa Fluor 647 Goat Anti-Rabbit IgG (H+L, 1:200; Invitrogen), Alexa Fluor 647 IgG (H+L) Cross-Adsorbed Goat anti-Human (1:200; Invitrogen), Alexa Fluor 488 goat anti-rabbit (1:200; Invitrogen), Texas Red-X goat anti-mouse IgG (1:200; Invitrogen) and Alexa Fluor 488 Phalloidin (to label F-actin, 1:1000; Invitrogen). For Western blotting, the following primary antibodies were used: anti-α-tubulin DM1A (1:5000; Sigma-Aldrich), anti-KIF18A (1:1000; Bethyl Laboratories Inc.), anti-CENP-A (3–19, 1:2000; Invitrogen) and anti-Histone H3 (3H1, 1:1000; New England BioLabs).
Normandin K., Coulombe-Huntington J., St-Denis C., Bernard A., Bourouh M., Bertomeu T., Tyers M, & Archambault V. (2023). Genetic enhancers of partial PLK1 inhibition reveal hypersensitivity to kinetochore perturbations. PLOS Genetics, 19(8), e1010903.
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2

Western Blot Analysis of Cellular Proteins

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Cells expressing individual sgRNAs were induced in 1 μg/mL
doxycycline for 2 to 5 days before lysis in Laemmli buffer and incubation at
95°C for 5 min. For mitotic samples, cells were harvested by mitotic
shake off and, when necessary, after an overnight 10 μM STLC
incubation. Samples were separated by SDS-PAGE and semi-dry transferred to
nitrocellulose. Membranes were blocked for 30 min in blocking buffer (5% BSA
for H2A.X; for all others, milk in TBS with 0.1% Tween-20) before incubation
with primary antibodies: anti-phospho-H2A.X (Ser139, Millipore clone JBW301;
1:1000), anti c-Myc (Abcam, ab32072; 1:1000), anti-CENP-A (Clone
3–19, Invitrogen; 1:2000), or
anti-“Bonsai”/NDC8093 (link) (0.5 μg/mL). This was
followed by HRP-conjugated secondary antibody (Kindle Biosciences)
incubation at 1:1000 dilution. To detect GAPDH as a loading control,
HRP-conjugated antibody (Abcam, ab185059) was applied at 1:20,000 dilution.
Membranes were imaged with a KwikQuant Imager (Kindle Biosciences) and
quantified using Image Studio software (LI-COR).
Funk L., Su K.C., Ly J., Feldman D., Singh A., Moodie B., Blainey P.C, & Cheeseman I.M. (2022). The phenotypic landscape of essential human genes. Cell, 185(24), 4634-4653.e22.
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