Ab129085

Manufactured by Abcam

Sourced in United Kingdom

Ab129085 is a lab equipment product. It is a recombinant antibody specific for an undisclosed target. The core function of this product is to bind to and detect the target antigen.

Automatically generated - may contain errors

Lab products found in correlation

Ab73287, by Abcam (1 mentions) Laemmli sample buffer, by Bio-Rad (1 mentions) Clarity western ecl substrate, by Bio-Rad (1 mentions) Ab195268, by Abcam (1 mentions) Polyvinylidene fluoride (pvdf), by Merck Group (1 mentions) Enhanced chemiluminescence western blotting system, by Thermo Fisher Scientific (1 mentions) A5441, by Merck Group (1 mentions) Protease inhibitor cocktail, by Roche (1 mentions) Ab192237, by Abcam (1 mentions) Ab17056, by Abcam (1 mentions) Ab124733, by Abcam (1 mentions) Ab174304, by Abcam (1 mentions)

3 protocols using ab129085

Protein Expression Analysis in Oocytes

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Oocytes were collected in 0.1% PVP in DPBS and boiled for 3 min at 95°C in 1× Laemmli Sample Buffer (Bio-Rad, 161-0747) supplemented with β-mercaptoethanol, proteinase inhibitor and phosphatase inhibitor. Samples were resolved on a 10% Laemmli gel and transferred onto supported nitrocellulose membranes, 0.2 μm (Bio-Rad, 1620097). Membranes were incubated in 5% blocking buffer for 1 h then incubated for 18 h in primary antibody at 4°C. The antibodies used: 1:1000 rabbit anti-CPEB1 (Abcam, ab73287), 1:1000 rabbit anti-CDC27 (Abcam, ab129085), 1:1000 mouse anti-CCNB1 (Abcam, ab72), 1:1000 mouse anti-STAT3 (Cell signaling, #3139), 1:1000 goat anti-CCNB2 (R&D Systems, AF6004), 1:10 000 rabbit anti-CPEB1 (Abcam, ab181051) and 1:1000 rabbit anti-ZAR1 (Bioss, bs-13549R). The membrane was then washed in 1× TBST, incubated in the appropriate secondary antibodies, 1:10 000 anti-rabbit IgG (GE Healthcare, NA934V), 1:10 000 anti-mouse IgG (GE Healthcare, NA931V), and 1:10 000 anti-goat IgG (TermoFisher, 31400) for 2 h, and washed again in 1× TBST. Clarity Western ECL substrate (Bio-Rad, 1705061) was then used to develop the membrane. All washes were done four times each round in TBST for 10 min each wash.

Luong X.G., Daldello E.M., Rajkovic G., Yang C.R, & Conti M. (2020). Genome-wide analysis reveals a switch in the translational program upon oocyte meiotic resumption. Nucleic Acids Research, 48(6), 3257-3276.

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Western Blot Analysis of Cell Proteins

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Western blotting was performed according to standard procedures. Briefly, total protein was extracted from the cells with lysis solution (10 mM Tris – HCl pH 7.4, 1 mM Na₂- EDTA, 0.1 % SDS, 10 mM Na-Vanadate, 1 mM PMSF and 1 X protease inhibitor cocktail (Complete, Roche, Mannheim, Germany). Aliquots (50 μg) of total protein extracts were resolved by electrophoresis in sodium dodecyl sulfate 7% polyacrylamide gels (SDS-PAGE) and electroblot transferred to polyvinylidene fluoride membranes (PVDF, Millipore). The membranes were then incubated with antibodies against BUB1 (1:1000; #ab195268, Abcam, Cambridge, United Kingdom), CDC27 (1:1000; #ab129085, Abcam) and β-Actin (1:20000, A5441, Sigma-Aldrich). Subsequently, the membranes were incubated with either anti-rabbit (1:1000; #7074, Cell Signaling Technology, Danvers, MA, USA) or anti-mouse (1:10000; #7076, Cell Signaling Technology) HRP-linked antibodies. Signals were detected using the enhanced chemiluminescence western blotting system (ThermoFisher Scientific). Densitometry was performed using Image Studio Lite Software version 5.2.5.

Wu J., Ferragut Cardoso A.P., States V.A., Al-Eryani L., Doll M., Wise S.S., Rai S.N, & States J.C. (2019). Overexpression of hsa-miR-186 induces chromosomal instability in arsenic-exposed human keratinocytes. Toxicology and applied pharmacology, 378, 114614.

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Immunoblot Analysis of Cell Cycle Proteins

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For immunoblotting, cells were grown in 60 mm cell culture dishes and whole cell extracts were obtained by lysis in radioimmunoprecipitation assay (RIPA) buffer (50 mM Tris–HCl (pH 8.0), 150 mM NaCl, 1.0% Igepal CA-630, 0.1% SDS, 0.1% Na-deoxycholic acid, supplemented with protease and phosphatase inhibitors) containing Benzonase (Novagen) and analysed by SDS–polyacrylamide gel electrophoresis following standard procedures. Primary antibodies were incubated over night at 4 °C in PBS-Tween containing 5% powder milk. Secondary peroxidase-coupled antibodies (Vector labs) were incubated at room temperature for 1 h. ECL-based chemiluminescene was detected with an Odyssee-Fc system.
Antibodies used for WB are: mouseTubulin (1:1,000, Santa Cruz, sc-8035), mouse MCM2 (1:500, Novus Biologicals, H00004171-M01), rabbit CDC27 (1:1,000, Abcam, ab129085), rabbit KIF23(MKLP1) (1:1,000, Abcam, ab174304), mouse PLK1 (1:1,000, Abcam, ab17056), rabbit CENPE (1:1,000, Abcam, ab124733), rabbit ANLN (1:1,000, Abcam ab99352) and rabbit CDCA5 (1:1,000, Abcam ab192237).

S. Pedersen R., Karemore G., Gudjonsson T., Rask M.B., Neumann B., Hériché J.K., Pepperkok R., Ellenberg J., Gerlich D.W., Lukas J, & Lukas C. (2016). Profiling DNA damage response following mitotic perturbations. Nature Communications, 7, 13887.

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