Nuclear protein extraction kit

Manufactured by Solarbio

Sourced in China

The Nuclear Protein Extraction Kit is a laboratory tool designed to isolate and extract nuclear proteins from cells or tissues. It provides a standardized and efficient method for the separation of nuclear and cytoplasmic fractions, enabling the study of nuclear-localized proteins and their interactions.

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Lab products found in correlation

Ripa lysis buffer, by Beyotime (4 mentions) Bca protein assay kit, by Solarbio (4 mentions) Pvdf membrane, by Merck Group (3 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions) Anti tubulin, by PTM Biolabs (3 mentions) Ab290, by Abcam (3 mentions) Ripa lysis buffer, by Solarbio (3 mentions) Histone h3, by Cell Signaling Technology (2 mentions) Ripa buffer, by Solarbio (2 mentions) Ripa buffer, by Beyotime (2 mentions) Anti gfp, by Abcam (2 mentions) Anti h3, by Abcam (2 mentions) Ab1791, by Abcam (2 mentions) Dna content quantitation assay, by Solarbio (2 mentions) Gapdh, by Cell Signaling Technology (2 mentions) Gapdh, by Abcam (2 mentions) Gapdh, by Proteintech (2 mentions) Gel pro analyzer 4, by Media Cybernetics (1 mentions) Confocal microscopy, by Olympus (1 mentions) Ab108595, by Abcam (1 mentions)

Close spelling variants of this product

Protein extraction kit (57 citations) Nuclear extraction kit (22 citations)

67 protocols using nuclear protein extraction kit

Subcellular Localization of BRAT1 Variants

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The GFP-tagged BRAT1 WT and R78P plasmids were transfected into 293T cells, respectively. After 48 h, the cells were washed with PBS and incubated with Hoechst 33342 for 5 min. Then, the Hoechst 33342 was removed and cells were washed with PBS for three times and imaged by confocal microscopy (Olympus) immediately.
Nuclear Protein Extraction kit Forty-eight hours after transfection of 1.5 μg plasmids in a 12-well plate, 293T cells were washed with PBS buffer and then treated with Nuclear Protein Extraction kit (R0050, Solarbio), according to the manufacturer's instructions. The products were collected and analyzed by Western blotting. GFP tag mouse monoclonal antibody (66002-1-Ig, Proteintech), antibody against GAPDH (10494-1-AP, Proteintech), and antibody against Lamin A/C (ab108595, Abcam) were used. The bands were calculated by Gel-Pro analyzer 4 (Media Cybernetics, L.P.).

Li W., Wu S., Xu H., Zhao X., Pan Y., Huang H., Lv H., Zhu X, & Liu Y. (2022). Novel variant in BRAT1 with the lethal neonatal rigidity and multifocal seizure syndrome. Pediatric research, 91(3).

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Quantification of SIRT1 Activity

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Nuclear extracts were prepared from fresh liver tissues with a Nuclear Protein Extraction Kit (Solarbio, Beijing, China). The protein concentration of nuclear extracts was quantified. A specific deacetylase detection kit was then used to determine SIRT1 activity of the nuclear extracts, as the procedures of the manufacturer (Genmed Scientifics Inc., Shanghai, China).

Chen Y., Li Y., Jia P., Ji S., Zhang H, & Wang T. (2022). Polydatin Attenuates Intra-Uterine Growth Retardation-Induced Liver Injury and Mitochondrial Dysfunction in Weanling Piglets by Improving Energy Metabolism and Redox Balance. Antioxidants, 11(4), 666.

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Western Blot Analysis of Proteins

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Total cell lysates were prepared in RIPA lysis buffer (Beyotime Biotechnology) supplemented with 1 mM PMSF and 1× protease inhibitor cocktail (Yeasen), and nuclear protein was extracted by nuclear protein extraction kit (Solarbio) in accordance with the manufacturer’s instructions. The samples were separated by SDS-PAGE and transferred to the PVDF membrane (Millipore). The membranes were incubated with the indicated primary antibodies at 4 °C overnight and then incubated with an HRP-conjugated anti-rabbit IgG antibody (Cell Signaling, Cat# 7074, 1:5000) or anti-mouse IgG antibody (Cell Signaling, Cat# 7076, 1:5000) at room temperature for 1 h. The signal was visualized by using the NcmECL Ultra kit (New Cell & Molecular Biotech) and then was exposed by using MiniChemi (SageCreation).

Wang X., Wang Y., Cao A., Luo Q., Chen D., Zhao W., Xu J., Li Q., Bu X, & Quan J. (2023). Development of cyclopeptide inhibitors of cGAS targeting protein-DNA interaction and phase separation. Nature Communications, 14, 6132.

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Cytosolic and Nuclear Protein Extraction

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K/G cells were washed once with PBS and centrifuged by using Legend Micro 17R centrifuge (Thermo Scientific) at 500 × g, 4°C for 2–3 min. The supernatant was aspirated and the pellet was retained. A total of 200 µl cytosol protein extraction kit (Beijing Solarbio Science & Technology Co., Ltd)/20 µl cell pellet (~2×10⁶ cells) was added, placed on ice for 10 min after pipetting evenly and centrifuged at 12,000 g and 4°C for 10 min. The supernatant contained plasma protein. The remaining precipitate was added to 50–100 µl nuclear protein extraction kit (Beijing Solarbio Science & Technology Co., Ltd). the procedure used to extract plasma proteins was used to extract nucleoprotein; the obtained supernatant contained nucleoprotein, which was used for subsequent experiments.

Wang X., Rong S., Sun Y., Yin B., Yang X., Lu X., Sun H., Yan Y., Sun G., Liang Y., Wang P., Xie S, & Li Y. (2023). NDRG3 regulates imatinib resistance by promoting β‑catenin accumulation in the nucleus in chronic myelogenous leukemia. Oncology Reports, 50(2), 152.

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Nuclear Protein Extraction and β-Catenin Immunoprecipitation

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According to the requirements of the reagent manufacturer, the nuclear protein extraction kit (Beijing Solarbio Science & Technology Co., Ltd) was used to extract the nucleoprotein, and the cells in a 10 cm culture dish need 800-1,000 µl the lysate. and centrifuged at 12,000 g for 10 min at 4°C. Nuclear extracts were incubated on a rotator with 8–10 µl anti-β-catenin overnight at 4°C. The anti-β-catenin antibody was used for protein blotting (1:100; cat. no. A19657; ABclonal Biotech Co, Ltd). A total of ~10 µl protein A/G agarose beads (cat. no. PR40025; Proteintech Group, Inc.) was added and the IgG control was detected by western blotting. The washing buffer was Tris-Buffered Saline and 0.1% Tween20 (TBST). The protein-antibody protein A/G-agarose bead complexes were boiled to achieve separation. Western blot verification was then carried out.

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Cellular Protein Extraction and Analysis

Cited 1 time

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Total proteins were prepared from cultured cell samples by complete cell lysis (Roche, Mannheim, Germany) with protease and phosphatase inhibitors. Nuclear proteins were prepared by Nuclear Protein Extraction Kit (Solarbio, Beijing, China) following the instruction from the manufacturer. Denatured proteins were separated on SDS-PAGE and transferred to membranes, followed by immunoblotting using antibodies of CypB (Abcam #ab16045), STAT3 (Cell Signaling Technology, #9139, #12640), pSTAT3 Tyr 705 (Cell Signaling Technology, #9145), SOCS3 (Cell Signaling Technology, #52113), β-actin (Sigma-Aldrich) and Histone H3 (Cell Signaling Technology, #4499). The bands were scanned and quantified as described [52 (link)].

Guo H., Zhuang K., Ding N., Hua R., Tang H., Wu Y., Yuan Z., Li T, & He S. (2022). High-fat diet induced cyclophilin B enhances STAT3/lncRNA-PVT1 feedforward loop and promotes growth and metastasis in colorectal cancer. Cell Death & Disease, 13(10), 883.

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Protein Expression Analysis of TICs

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Each group's fresh ovarian tissue was removed, and the P1 generation of TICs was cultivated until it reached 90% confluence. Nuclear protein extraction kit (Solarbio, Beijing, China)was used to extract nuclear and cytoplasmic protein from TICs as well as total protein using RIPA pyrolysis. SDS-PAGE was used to separate the proteins, and they were then moved to the polyvinylidene difluoride membrane (PVDF). The membranes were incubated with the primary antibodies Cyp17a1 (1:4000 dilution; ab125022; Abcam), NR4A1 (1:500 dilution; sc-166166; Santa Cruz), Phospho-NR4A1 (1:1000 dilution; Ser351; Cell signaling technology), Bcl-2 (1:1000 dilution; Abclonal), Bax (1:1000 dilution; Abclonal), Caspase-9(1:1000 dilution; Abclonal), caspase-3(1:500 dilution; ab32351; Abcam) and cytc(1:500 dilution; Cell signaling technology) at 4℃ overnight, following being washed three times with TBS added with Tween 20(TBST). The second antibody was administered at room temperature for one hour and was HRP-conjugated affinipure goat anti-rabbit IgG (H + L) (1:40,000 dilution; 10,285–1-AP; Proteintech). Utilizing the Tanon 5200 analytical equipment (Tanon, Shanghai, China), chemiluminescence was measured following reaction with the Ultra-sensitive chemiluminescence kit (Sparkjade, Qingdao, Shandong).

Luo Q., Tang Y., Jiang Z., Bao H., Fu Q, & Zhang H. (2022). hUCMSCs reduce theca interstitial cells apoptosis and restore ovarian function in premature ovarian insufficiency rats through regulating NR4A1-mediated mitochondrial mechanisms. Reproductive Biology and Endocrinology : RB&E, 20, 125.

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Protein Extraction from Vascular Smooth Muscle Cells

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At collection time points, culture medium was removed and then VSMCs were washed with 1× PBS for 3 times. The nuclear and cytoplasmic protein lysate extraction of VSMCs was performed using the Nuclear Protein Extraction Kit (Solarbio) according to the manufacturer’s recommendations.

Li W., Feng W., Su X., Luo D., Li Z., Zhou Y., Zhu Y., Zhang M., Chen J., Liu B, & Huang H. (2022). SIRT6 protects vascular smooth muscle cells from osteogenic transdifferentiation via Runx2 in chronic kidney disease. The Journal of Clinical Investigation, 132(1), e150051.

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Nuclear Protein Extraction and Western Blot Analysis

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Cells after treatment were washed twice with PBS and nuclear proteins were extracted from cells using nuclear protein extraction kit (Solarbio). Then, equal amounts of proteins were subjected to 8–12% SDS-PAGE and western blot analysis. The primary antibodies used in this study: anti-Nrf2 Ab (16396-1-AP, 1 : 1000; Proteintech) and anti-Lamin A + C (ab169532, 1 : 5000; Abcam). Anti-Lamin A + C antibodies were used for equal loading of nuclear proteins.

Geng J., Ren Q., Chang C., Xie X., Liu J, & Du Y. (2019). Synthesis and biological activities of petrosiols B and D. RSC Advances, 9(18), 10253-10263.

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Regulatory Mechanisms of Nrf2 and AKT in Cancer Therapy

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Anti-Nrf2 (ab89443) purchased from Abcam (Cambridge, UK). Anti-β-actin (20536-1-AP), anti-BAD (10435-1-AP), p-CREB (12208-1-AP), LMNA (10298-1-AP) antibodies were obtained from Proteintech Group Co., Ltd. (Wuhan, PRC). Anti-AKT1(K101311P), anti-p-AKT1(K006214P), PI3K(K106692P), p-PI3K(K006379P) antibodies were obtained from Solarbio Science & Technology(Beijing, PRC). Anti-BCL-2(AF6139), Cleaved-Caspase 3(AF7022), Caspase 3(AF6311), Cleaved-Caspase 9(AF5240), Caspase 9(AF6348) antibodies were obtained from, Affinity Biosciences (USA). Vincristine (VCR) were purchased from Topscience (Shanghai, PRC). Daunorubicin (DNR) and brusatol (an Nrf2 inhibition) were purchased from MCE (NJ, USA). The chemical inhibitor MK-2206 (MCE, NJ, USA) of the AKT (39 (link)). Fetal bovine serum and RPMI 1640 medium were obtained from Gibco (Carlsbad, CA, USA). Nuclear Protein Extraction Kit were obtained from Solarbio Science & Technology (Beijing, PRC).

Wang L., Liu X., Kang Q., Pan C., Zhang T., Feng C., Chen L., Wei S, & Wang J. (2022). Nrf2 Overexpression Decreases Vincristine Chemotherapy Sensitivity Through the PI3K-AKT Pathway in Adult B-Cell Acute Lymphoblastic Leukemia. Frontiers in Oncology, 12, 876556.

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