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Anti cenpa

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Anti-CENPA is a primary antibody that specifically recognizes the CENPA protein, a key component of the centromere-specific chromatin complex. This antibody can be used for the detection and analysis of CENPA in various applications such as Western blotting, immunohistochemistry, and immunocytochemistry.

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Lab products found in correlation

Anti nucleolin, by Abcam (2 mentions) Anti gfp, by Abcam (2 mentions) Alexa 594 labeled secondary antibody, by Thermo Fisher Scientific (1 mentions) Dapi staining mounting medium, by Vector Laboratories (1 mentions) Fluorescence microscope, by Olympus (1 mentions) Anti nanog, by Abcam (1 mentions) Anti fibrillarin, by Abcam (1 mentions) Anti sma, by Merck Group (1 mentions) Anti β actin, by Santa Cruz Biotechnology (1 mentions) Anti map2, by Merck Group (1 mentions) Anti tuj1, by Merck Group (1 mentions) Anti nestin, by Merck Group (1 mentions) Anti neun, by Merck Group (1 mentions) Anti alb, by Abcam (1 mentions) Anti cd105, by Thermo Fisher Scientific (1 mentions) Anti caldesmon, by Merck Group (1 mentions) Anti igg apc, by Thermo Fisher Scientific (1 mentions) Anti sm22, by Abcam (1 mentions) Anti γh2ax, by Merck Group (1 mentions) Anti sox2, by Santa Cruz Biotechnology (1 mentions)

7 protocols using anti cenpa

1

Immunofluorescence Staining of HA-SENP1, HA-SENP2, Endogenous SENP2, PIAS4, and CENP-A in U2OS Cells

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For HA-SENP1, HA-SENP2 staining, cultured U2OS cells were transient transfected with above HA tagged plasmids. Cells were stained with anti-HA antibody (cat. #H6908, Sigma). For endogenous SENP2 staining, cells were stained with anti-SENP2 antibody (cat. # ab58418, Abcam). For endogenous PIAS4 and CENP-A staining, U2OS cells were stained with anti-PIAS4 (cat. # ab211625, Abcam) and anti-CENP-A (cat. #2186, Cell Signaling). After incubation with the primary antibodies at room temperature for 1 hr, Alexa 488- and Alexa 594-labeled secondary antibodies (Life Technology) were added for 1 hr. Cells were then adhered to a slide with DAPI-staining Mounting Medium (Vector Labs). All samples were visualized with an Olympus fluorescence microscope and images were derived with the accompanying DP-BSW application software program.
Wang R., Liu F., Zhao Y., Wu D., Chen L., Yeh E.T, & Huang C. (2017). Reversible regulation of ORC2 SUMOylation by PIAS4 and SENP2. Oncotarget, 8(41), 70142-70155.
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2

Comprehensive Antibody Characterization Protocol

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The following antibodies were used: anti-ALB (Abcam, ab8940, 1:400); anti-β-actin (Santa Cruz Biotechnology, sc-130301, 1:5 000); anti-caldesmon (Sigma-Aldrich, C4562, 1:300); anti-CD73 (BD Biosciences, 550741, 1:50); anti-CD90 (BD Biosciences, 555595, 1:100); anti-CD105 (eBioscience, 1-1057, 1:100); anti-CENPA (Abcam, ab13939, 1:400); anti-fibrillarin (Abcam, ab4566, 1:100); anti-FLAG (Sigma-Aldrich, M2, 1:2 000 for western blotting, 1:400 for immunofluorescence); anti-GAL4 (Abcam, ab14477, 1:1 000 for western blotting, 1:400 for immunofluorescence); anti-γH2AX (Millipore, 05-636, 1:400); anti-IgG-APC (eBioscience, 555751, 1:100); anti-IgG-FITC (eBioscience, 555748, 1:100); anti-IgG-PE (eBioscience, 555749, 1:100); anti-MAP2 (Sigma-Aldrich, m4403, 1:500); anti-NANOG (Abcam, ab21624, 1:250); anti-nestin (Millipore, MAB5326, 1:500); anti-NeuN (Millipore, ABN78 1:400); anti-OCT4 (Santa Cruz Biotechnology, sc-5279, 1:100); anti-Nucleolin (Abcam, ab22758, 1:200); anti-PAX6 (Covance, PRB-278P, 1:500); anti-SMA (Sigma-Aldrich, A5228, 1:100); anti-SM22 (Abcam, ab14106, 1:200); anti-SOX2 (Santa Cruz Biotechnology, sc-17320, 1:100); anti-Tuj1 (Sigma-Aldrich, T2200, 1:500); Alexa Fluor 555-conjugated wheat germ agglutinin (Thermo Fisher, W32464, 1:500). Other reagents, including deoxycytidine, hydroxyurea, NAC, nocodazole, and thymidine, were purchased from Sigma-Aldrich.
Ren R., Deng L., Xue Y., Suzuki K., Zhang W., Yu Y., Wu J., Sun L., Gong X., Luan H., Yang F., Ju Z., Ren X., Wang S., Tang H., Geng L., Zhang W., Li J., Qiao J., Xu T., Qu J, & Liu G.H. (2017). Visualization of aging-associated chromatin alterations with an engineered TALE system. Cell Research, 27(4), 483-504.
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3

Indirect Immunofluorescence Imaging of Cellular Proteins

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For indirect immunofluorescence, cells plated onto poly-L-lysine-coated coverslips were fixed in 4% formaldehyde for 10 mins. Cells were pre-extracted with 0.3% Triton X-100 in PBS for 5 mins and incubated in Triton Block (0.2 M Glycine, 2.5% FBS, 0.1% Triton X-100, PBS). The following primary antibodies were used at 1:1,000 dilution (unless noted) in Triton Block and washed with 0.1% Triton X-100 in PBS: anti-CENP-A (Abcam, ab13939), anti-CENP-C (MBL, PD030), 1:400 ACA (Antibodies Incorporated, 15-235-0001), anti-phospho H2AX (S139) clone JBW301 (EMD Millipore, 05-636), and 1:200 anti-Lamin B1 (Proteintech Group, 12987-1-ap). Immunofluorescent images were acquired on a DeltaVision Core system at 40–60× magnification (30 × 0.2μm z-sections) and deconvolved maximum intensity projections were generated using softWoRx program. ImageJ was used to quantify fluorescent intensity. For immunofluorescence combined with DNA FISH, the immunofluorescence procedure was performed first followed by the described FISH procedure.
Ly P., Teitz L.S., Kim D.H., Shoshani O., Skaletsky H., Fachinetti D., Page D.C, & Cleveland D.W. (2016). Selective Y centromere inactivation triggers chromosome shattering in micronuclei and repair by canonical NHEJ. Nature cell biology, 19(1), 68-75.
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4

Immunofluorescence Assay for Centromeric Proteins

Cited 4 times
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IF on settled interphase cells and metaphase spreads were performed as previously described (Chen et al., 2014 (link)). Primary antibodies: anti-CENP-A (chicken, 1:1,500; Blower and Karpen, 2001 (link)) or anti-CID (rabbit, 1:500; Abcam), anti-CENP-C (guinea pig, 1:500; Erhardt et al., 2008 (link)), anti-Ndc80 (chicken, 1:200; Cane et al., 2013 (link)), anti-GFP Alexa 488-conjugated (rabbit, 1:100; Invitrogen), or anti-GFP (chicken, 1:500; Abcam), and anti-HA (mouse, 1:500; Covance).
For salt extractions, settled cells were incubated with PBS-D (0.1% digitonin) with or without 0.5 M NaCl for 30 min (Perpelescu et al., 2009 (link)) before 37% formaldehyde was added to the solution to a final concentration of 3.7% followed by 10 min of incubation before proceeding with IF.
Rosin L, & Mellone B.G. (2016). Co-evolving CENP-A and CAL1 Domains Mediate Centromeric CENP-A Deposition across Drosophila Species. Developmental cell, 37(2), 136-147.
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5

Antibody Panel for Nuclear Protein Analysis

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The following primary antibodies were used in this study: anti-CENPA (AB13939; Abcam), anti-SC35 (S4045; MilliporeSigma), anti-MFAP1 (HPA042370; MilliporeSigma), anti-SRRM2 (PA5-68009; Thermo Fisher Scientific), anti-ZNF207 (HPA017013; MilliporeSigma), anti-HP1α (H2164; MilliporeSigma), anti-β Tubulin (AB18207; Abcam), anti-FITC (200-002-037; Jackson ImmunoResearch), custom-made anti-SON (Chen et al., 2018 (link); PACIFIC10700; Pacific Immunology), anti-Nucleophosmin (AB86712; Abcam), anti-nucleolin (AB70493; Abcam), anti-Coilin (Ab87913; Abcam), and anti-PRPF38A (PA5-62730; Thermo Fisher Scientific). Secondary antibodies used were goat anti-mouse HRP (115-035-062; Jackson ImmunoResearch), goat anti-rabbit HRP (111-035-144; Jackson ImmunoResearch), goat anti-mouse FITC (111-095-144; Jackson ImmunoResearch), and goat anti-rabbit Texas red (115–075-146; Jackson ImmunoResearch).
Dopie J., Sweredoski M.J., Moradian A, & Belmont A.S. (2020). Tyramide signal amplification mass spectrometry (TSA-MS) ratio identifies nuclear speckle proteins. The Journal of Cell Biology, 219(9), e201910207.
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6

Immunofluorescence Staining of DNA Repair Proteins

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Scramble and USP24-knockdown lentivirus infected cells were seeded in 6-well plates with cover slips inside for 48 h. Cover slips were removed and cells were fixed with 4% paraformaldehyde in 4 °C for 15 min. After fixation, cover slips were washed with PBS, and incubated with 0.2% Triton X-100 in PBS for 5 min at room temperature. Cover slips were then blocked with 1% BSA for 1 h, and stained with anti-Rad51 antibody (1:200, Genetex), anti-GFP (santa cruz), anti-CENPA (abcam), or anti-γ-H2Ax (abcam) for 16 h at 4 °C. After washing with PBS, cells were stained with Alexa Fluor® 488 or 568 (Invitrogen) for 1 h at room temperature, and mounted with 90% glycerol containing DAPI (Invitrogen). Number of stained cells, Rad51, and γ-H2Ax foci were examined by fluorescence microscopy (Olympus, Tokyo, Japan). ImageJ (Bethesda, Maryland, USA.) were used to perform the statistical analysis of Rad51 and γ-H2Ax foci.
Wang S.A., Young M.J., Wang Y.C., Chen S.H., Liu C.Y., Lo Y.A., Jen H.H., Hsu K.C, & Hung J.J. (2021). USP24 promotes drug resistance during cancer therapy. Cell Death and Differentiation, 28(9), 2690-2707.
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7

Cell Cycle Regulation Protein Antibodies

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The detailed procedure was described
in our previous study.54 (link),55 (link),58 (link),59 (link) Primary antibodies, anti-CENPA, anti-CENPE,
anti-CENPF, and anti-CENPI antibodies, were purchased from abcam (Cambridge,
U.K.); anti-CDK1 and anticyclin B1 antibodies were purchased from
Santa Cruz Biotechnology, Inc. (Dallas, TX); and anti-EZH2 and anti-H3K27me3 antibodies were purchased from GeneTex International Corporation
(HsinChu, Taiwan), anti-Caspase-3 was purchased from GeneTex International
Corporation; anti-cleaved Caspase-3 was purchased from Cell Signaling
Technology (Danvers, Massachusetts).
Sharma S., Wang S.A., Yang W.B., Lin H.Y., Lai M.J., Chen H.C., Kao T.Y., Hsu F.L., Nepali K., Hsu T.I, & Liou J.P. (2024). First-in-Class Dual EZH2-HSP90 Inhibitor Eliciting Striking Antiglioblastoma Activity In Vitro and In Vivo. Journal of Medicinal Chemistry, 67(4), 2963-2985.
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