For immunofluorescence of bone sections, the fixed bone sections were blocked in 0.8% bovine serum albumin (BSA) for 1 h. Sections were incubated with the primary antibodies (1:200) against CDC20 (Abcam; ab183479) overnight, followed by incubation in 1:500 secondary antibody (ZF-0311, ZSGB-BIO, Beijing, China) for 1 h. Nuclei were stained using 4 ′,6-diamidino-2-phenylindole (DAPI).
Ab183479
Manufactured by Abcam
Sourced in China, United Kingdom
Ab183479 is a monoclonal antibody used for the detection of a specific target protein. It is suitable for use in various laboratory techniques, such as Western blotting and immunohistochemistry. The antibody is produced in a mouse host and purified by protein A affinity chromatography.
Automatically generated - may contain errors
Lab products found in correlation
Zf 0311, by ZSGB-BIO (1 mentions)
Ab252940, by Abcam (1 mentions)
Ab52934, by Abcam (1 mentions)
Ab172476, by Abcam (1 mentions)
Ab6721, by Abcam (1 mentions)
Nocodazole, by Selleck Chemicals (1 mentions)
Ab81292, by Abcam (1 mentions)
Ab201022, by Abcam (1 mentions)
Ku55933, by Selleck Chemicals (1 mentions)
Ab70385, by Abcam (1 mentions)
Lambda protein phosphatase λppase, by Merck Group (1 mentions)
Sc 81514, by Santa Cruz Biotechnology (1 mentions)
Sc 9996, by Santa Cruz Biotechnology (1 mentions)
Sc 5267, by Santa Cruz Biotechnology (1 mentions)
F3165, by Merck Group (1 mentions)
Ab109315, by Abcam (1 mentions)
Ab207298, by Abcam (1 mentions)
Ab128245, by Abcam (1 mentions)
Ab32053, by Abcam (1 mentions)
Ab1791, by Abcam (1 mentions)
4 protocols using ab183479
1
Immunofluorescence of Bone Sections
2
Immunohistochemical Analysis of Cell Cycle Regulators
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Clinical tissues were fixed in 4% paraformaldehyde for 12 h, dewaxed in xylene, rehydrated in descending series of alcohol (100, 95 and 75% alcohol), and heated in 0.01 M citrate buffer for 15–20 min. After cooling to room temperature, the tissues were washed with PBS, blocked with goat serum and left to stand at room temperature for 20 min. Next, the tissues were immunostained with primary antibodies (Abcam, Cambridge, UK) against CDC20 (ab183479, 1:200), TOP2A (ab52934, 1:200), RRM2 (ab172476, 1:200), UBE2C (ab252940, 1:200) or AOX1 (ab197828, 1:200) at room temperature for 1 h. The tissues were incubated with 30 μL secondary goat anti-rabbit immunoglobulin G (ab6721, 1:2000, Abcam) for 1 h at room temperature. Thereafter, the tissues were treated with streptavidin-peroxidase, allowed to stand at 37°C for 30 min, developed with 3,3′-diaminobenzidine tetrahydrochloride for 5–10 min, counterstained for 2 min, and differentiated with hydrochloric acid-ethanol. The tissues were observed under a microscope after conventional dehydration, clearing and mounting.
3
Immunoblotting Analysis of Mitotic Regulators
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Nocodazole and Ku55933 were ordered from Selleckchem (Texas, USA). Lambda Protein phosphatase (λPPase) was bought from Sigma-Aldrich Corporation. The anti-ATM (1:1,000, ab201022, Abcam), anti-phospho-ATM (1:1,000, ab81292, Abcam), anti-Mad2 (1:1,000, ab70385, Abcam), anti-Mad1 (1:1,000, ab201022, Abcam), and anti-Cdc20 antibodies (1:1,000, ab183479, Abcam) were bought from Abcam (Cambridge, MA). The anti-HA (1:1,000, 3724T, Cell Signaling Technology), anti-Mad2 (1:1,000, 4636S, Cell Signaling Technology), and the HRP-conjugated secondary antibodies were purchased from Cell Signaling Technology (Danvers, MA). The anti-Flag antibody (1:1,000, F3165, Sigma) was purchased from Sigma. The anti-Mad2 antibody (1:1,000, YT2618, Immunoway) was bought from Immunoway. The anti-GFP (1:1,000, sc-9996, Santa Cruz), anti-Mad2 (1:1,000, sc-47747, Santa Cruz), anti-p-Thr (1:1,000, sc-5267, Santa Cruz), and anti-p-Ser antibodies (1:1,000, sc-81514, Santa Cruz) were bought from Santa Cruz Biotechnology. Goat anti-mouse lgG-H HRP (1:1,000, M21004L, Abmart) and goat anti-mouse lgG-L HRP (1:1,000, M21005S, Abmart) were purchased from Abmart.
4
Western Blot Protein Expression Analysis
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The media was discarded, and RIPA lysate was added to the cells for total protein extraction. The total protein concentration was measured using the BCA technique (Thermo, Shanghai, China). Protein samples were separated through SDS-PAGE and transferred onto a PVDF membrane. After blocking with 5% skim milk at room temperature for 1 h, the membrane was washed three times with TBST and then incubated overnight at 4 °C with the primary antibody at the recommended dilution ratio. Subsequently, the membrane was washed again with TBST and incubated at room temperature for 1 h with the DyLight 800-labeled secondary antibody (ROCKLAND, USA). The membrane underwent three additional TBST washes before imaging using a fluorescence scanning instrument (Odyssey, Danbury, CT, USA). Primary antibodies against FOXM1 (ab207298, dilution: 1/1,000), FAM83A (ab128245, dilution: 1/1,000), CDC20 (ab183479, dilution: 1/2,000), CDC6 (ab109315, dilution: 1/4,000), cyclin B (ab32053, dilution: 1/8,000), H3 (ab1791, dilution: 1/5,000), and GAPDH (ab9485, dilution: 1/2,500) were bought from Abcam for this work.
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