P0025

Manufactured by Beyotime

Sourced in China

P0025 is a benchtop centrifuge designed for laboratory use. It can be used to separate samples based on density differences. The centrifuge has a maximum speed of 6,000 rpm and a maximum relative centrifugal force (RCF) of 4,000 g. It is suitable for a variety of applications that require sample separation in a laboratory setting.

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Lab products found in correlation

4 protocols using p0025

Sequential Immunofluorescence Staining

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In this study, both anti-ADCY3 and anti-PDE3A are rabbit IgG antibodies, we performed immunofluorescence re-staining to observe their localization and expression in ovaries (34 (link)). After the immunofluorescence staining of ADCY3, the position of the section was recorded when taking photos. Subsequently, the original primary and secondary antibodies on the sections were eluted with stripping buffer (P0025, Beyotime Biotechnology, China), and then PDE3A was re-stained according to immunofluorescence staining protocols. Finally, photos were taken at the same location.

Zheng W., Zhang T., Zhao T., Zhu Z., Qin S., Yan H., He M., Zhou B., Xia G., Zhang H, & Wang C. (2023). cAMP controls the balance between dormancy and activation of primordial follicles in mouse ovaries. PNAS Nexus, 2(3), pgad055.

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Western Blot Analysis of Spinal Cord Proteins

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Total protein was extracted from the L4-L6 spinal cord segments as previously described66 (link). Proteins were separated by SDS-PAGE and transferred to PVDF membranes. The membranes were incubated overnight at 4 °C with the following primary antibodies: rabbit anti-GFAP (1:500) (ab53554, Abcam), rabbit anti-Substance P (1:500) (orb215527, Biorbyt), rabbit anti-CGRP (1:500) (ab47027, Abcam), rabbit anti-PDGF BB (1:300) (ab16829, Abcam), rabbit anti-pPDGFRα (1:1000) (#2992, Cell Signal), rabbit anti-pPDGFRβ (1:1000) (#2227,Cell signal), rabbit anti-ERK(1:500) (ab184699, Abcam), rabbit anti-pERK (1:1000) (#4370, Cell Signal), rabbit anti-AKT (1:500) (9272s, Cell Signal) and rabbit anti-pAKT (1:1000) (4056s, Cell Signal). The membranes were subsequently incubated with the appropriate secondary antibodies (1:5000; Abcam) for 1 h at room temperature. The protein bands were detected using an enhanced chemiluminescence detection system. Then, the membranes were stripped with stripping buffer (P0025, beyotime) and reblotted with antibodies against the loading controls [mouse anti-β-actin (1:5000) (ab119716, Abcam) and mouse anti-GAPDH (1:5000) (ab9484)]. The intensity of the immunoreactive bands was quantified using Image J software. The ratio of target protein to β-actin (loading control) levels was statistically analyzed. Western blot experiments were repeated 3 times.

Xu Y., Liu J., He M., Liu R., Belegu V., Dai P., Liu W., Wang W., Xia Q.J., Shang F.F., Luo C.Z., Zhou X., Liu S., McDonald J., Liu J., Zuo Y.X., Liu F, & Wang T.H. (2016). Mechanisms of PDGF siRNA-mediated inhibition of bone cancer pain in the spinal cord. Scientific Reports, 6, 27512.

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Comprehensive Protein Analysis in Cellular Signaling

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PI3K (ab86714, Abcam), AKT3 (bs-5146R, Bioss), mTOR (2972s, CST), LC3B (bs-2912R, Bioss), P62 (18420-1-AP, Triple Eagle), western removal buffer of primary antibody and second antibody (P0025, Beyotime), RIPA cell lysate (P0013B, Beyotime), ECL hypersensitive luminescence kit (34095, Thermo), goat anti-mouse IgG secondary antibody (ZB-2305, Zsbio), goat anti-rabbit IgG secondary antibody (ZB-2301, Zsbio), goat anti-rabbit IgG (FITC) (B029, Ebiogo), sheep serum block (B010, Ebiogo), anti-fluorescence quench blocking agent (containing DAPI) (B024, Ebiogo), TRIzol (15596026, Life Technologies), and hematoxylin (BA-4041, BaSO).
EPS 300 electrophoresis instrument (Tanon), VE-180 electrophoresis tank (Tanon), JW-3021HR high-speed refrigerated centrifuge with 6.8cm centrifugal radius (Anhui Jiawen Instrument Equipment), JEM1400 flash transmission electron microscope (Jieou Lu, Beijing), PIKOREAL 96 fluorescence quantitative PCR instrument (Thermo), OD1000+ ultra-micro spectrophotometer (Nanjing Wuyi), CX41 microscope (Olympus), RM2016 Leica microtome (Leica), Pannoramic MIDI digital section scanner (3DHISTECH), and a 1319A digital thermometric indicator(Shanghai TES).

Jia Y.M., Zhu C.F., She Z.Y., Wu M.M., Wu Y.Y., Zhou B.Y, & Zhang N. (2022). Effects on Autophagy of Moxibustion at Governor Vessel Acupoints in APP/PS1double-Transgenic Alzheimer's Disease Mice through the lncRNA Six3os1/miR-511-3p/AKT3 Molecular Axis. Evidence-based Complementary and Alternative Medicine : eCAM, 2022, 3881962.

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Western Blot Analysis of Cardiac Proteins

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Left ventricular myocardial tissue or cardiomyocytes were isolated and lysed with cold 1 × lysis buffer supplemented with a protease inhibitor cocktail (Thermo Fisher Scientific, Waltham, MA, USA). Protein concentration was measured using a BCA protein assay kit (Thermo Fisher Scientific, Waltham, MA, USA) and then mixed with 1 × SDS sample buffer and boiled at 95 °C for 5 min. The lysate was resolved by SDS-PAGE and transferred onto polyvinylidene difluoride (PVDF) membranes, which were blocked and incubated with primary antibodies at 4 °C overnight, followed by incubation with horseradish peroxidase (HRP)-conjugated secondary antibodies. The signals were detected using the ECL system (Pierce) and quantified by scanning densitometry using the Image Lab analysis system (see Supplementary Material for original western blots). The membranes were stripped with stripping buffer (P0025, Beyotime, Beijing, China) and reblotted with next primary antibody. Primary antibodies: anti-Peli1 (ab199336, Abcam, Cambridge, MA, USA), anti-HNF4α (ab41898, Abcam, Cambridge, MA, USA), anti-GAPDH (AF0006, Beyotime, Beijing, China), anti-Flag (F2555, Sigma-Aldrich, St. Louis, MO, USA), anti-Myc (AF2864, Beyotime, Beijing, China), anti-His (AF2876, Beyotime, Beijing, China), anti-HA (AF2858, Beyotime, Beijing, China).

Hou Y., Shi P., Du H., Zhu C., Tang C., Que L., Zhu G., Liu L., Chen Q., Li C., Shao G., Li Y, & Li J. (2024). HNF4α ubiquitination mediated by Peli1 impairs FAO and accelerates pressure overload-induced myocardial hypertrophy. Cell Death & Disease, 15(2), 135.

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