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P2012

Manufactured by Beyotime
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Sourced in China

The P2012 is a versatile laboratory equipment designed for general scientific applications. It serves as a reliable tool for performing various laboratory tasks. The core function of the P2012 is to provide a stable and controlled environment for experiments and analysis.

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Lab products found in correlation

Protein a g agarose, by Beyotime (2 mentions) Iqgap1 rabbit pab, by Santa Cruz Biotechnology (1 mentions) P8340, by Merck Group (1 mentions) Ab80579, by Abcam (1 mentions) P0013, by Beyotime (1 mentions) C abl rabbit polyclonal antibody, by Cell Signaling Technology (1 mentions)

6 protocols using p2012

1

Acetylation of OPA1 by Co-Immunoprecipitation

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For CO‐IP, 1 mg of protein in 100 μl of lysis buffer was pre‐cleared for 1 h at 4°C with protein A + G agarose (P2012; Beyotime Biotechnology). After that, rabbit anti‐SIRT3 (D22A3; Cell Signaling Technology) or mouse anti‐OPA1 (ab119685; Abcam) was added to the sample and incubated for overnight at 4°C. Then, the complex was added with 20 μl of protein A + G agarose at 4°C for 4 h. After incubation and washing, immunoprecipitates were boiled in SDS/PAGE loading buffer and subjected to Western blotting analysis. To detect the acetylated OPA1, the protein was incubated with 1 μg of anti‐OPA1 overnight, and then, the samples were measured using the rabbit antibody of acetylated lysine (9441; CST) by Western blotting.
Yuan L., Yang J., Li Y., Yuan L., Liu F., Yuan Y, & Tang X. (2022). Matrine alleviates cisplatin‐induced acute kidney injury by inhibiting mitochondrial dysfunction and inflammation via SIRT3/OPA1 pathway. Journal of Cellular and Molecular Medicine, 26(13), 3702-3715.
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2

Co-immunoprecipitation of Nephrin and IQGAP1

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Co-immunoprecipitation experiments were conducted according to the manufacturer’s instructions (P2012, Beyotime, China). The total protein of the cultured podocytes or COS7 cells was extracted using lysis buffer (1.0% Triton X-100, 150 mM NaCl, 5 mM EDTA, 1 mM PMSF, 20 mM Tris, pH 7.5) that contained a protease inhibitor cocktail (P8340, Sigma-Aldrich, USA). The samples were centrifuged at 13,000 g for 5 min at 4 °C. Nephrin rabbit pAb (2 μg/500 μg total protein; Santa Cruz, USA) or IQGAP1 rabbit pAb (2 μg/500 μg total protein, Santa Cruz, USA) was added to the supernatant and rotated overnight at 4 °C. Then, the mixture was loaded with protein A + G agarose and incubated for 3 h at 4 °C. The centrifuged sediment was saved and mixed with 1× LDS sample buffer. After boiling at 70 °C for 10 min, the samples were analyzed by Western blotting.
Liu Y., Liang W., Yang Y., Pan Y., Yang Q., Chen X., Singhal P.C, & Ding G. (2015). IQGAP1 regulates actin cytoskeleton organization in podocytes through interaction with nephrin. Cellular signalling, 27(4), 867-877.
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3

Tau5 Immunoprecipitation from Virus-Infected Hippocampal Tissue

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Hippocampal regions infected with virus were isolated and mechanically homogenized in lysis buffer for immunoprecipitation (IP) (P0013, Beyotime). Then, the homogenate was centrifuged at 3,000 rpm, for 20 min at 4°C. The supernatant was incubated with the primary antibody Tau5 (2 µg/100 µg) (ab80579, Abcam) overnight at 4°C and then Protein A+G Agarose (30 µl/100 µl) (P2012, Beyotime) was added into the sample for 4–6 h. After that, the agarose was washed three times. Proteins attached to the agarose were resuspended in buffer (50 mM Tris-HCl, pH 6.8, 2% SDS, 10% glycerol) and boiled for 10 min. Collected protein sample was analyzed by Western blot.
Jiang X.J., Wu Y.Q., Ma R., Chang Y.M., Li L.L., Zhu J.H., Liu G.P, & Li G. (2022). PINK1 Alleviates Cognitive Impairments via Attenuating Pathological Tau Aggregation in a Mouse Model of Tauopathy. Frontiers in Cell and Developmental Biology, 9, 736267.
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4

Protein Interactions: c-Abl, Nephrin, and SHIP2

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Coimmunoprecipitation experiments were performed according to the manufacturer’s instructions (P2012; Beyotime, China). The total proteins from the cultured podocytes were extracted using lysis buffer (20 mM Tris, 150 mM NaCl, 1.0% Triton X-100, 5 mM EDTA, and 1 mM phenylmethylsulfonyl fluoride, pH 7.5). A c-Abl rabbit polyclonal antibody (1:200; Cell Signaling Technology) was added to the protein samples, and these were then rotated overnight at 4°C. The mixture was then loaded with 40 μl of protein A+G–agarose, incubated for 3 h at 4°C, centrifuged at 2500 × g and 4°C for 5 min and washed five times with phosphate-buffered saline (PBS). The beads were mixed with 1× Lane Marker Sample Buffer. After being boiled at 95–100°C for 5 min, the samples were analyzed by Western blotting for c-Abl, nephrin, and SHIP2 expression.
Yang Q., Ma Y., Liu Y., Liang W., Chen X., Ren Z., Wang H., Singhal P.C, & Ding G. (2016). Angiotensin II down-regulates nephrin–Akt signaling and induces podocyte injury: roleof c-Abl. Molecular Biology of the Cell, 27(1), 197-208.
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5

NEK7-NLRP3 Interaction by Co-IP

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For detecting the interaction of NEK7 and NLRP3 by co-immunoprecipitation (Co-IP). According to the manufacturer's instructions (P2012, Beyotime Biotechnology, China), the centrifuged lysates were incubated with a 1:200 dilution of anti-NEK7 or anti-NLRP3 antibody overnight, at 4°C. Then, 40 µl Protein A/G Agarose (Beyotime, China) was added and incubated for an additional 4 h in a shaker. The immune complexes were boiled in the sample buffer after washing with PBS ve times. The samples were then immunoblotted with anti-NLRP3 or anti-NEK7 respectively.
Liang L., Wang H., Hu Y., Bian H., Xiao L, & Wang G. (2022). Oridonin relieves depressive-like behaviors by inhibiting neuroinflammation and autophagy impairment in rats subjected to chronic unpredictable mild stress. Phytotherapy research : PTR, 36(8).
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6

NEK7-NLRP3 Interaction by Co-IP

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For detecting the interaction of NEK7 and NLRP3 by co-immunoprecipitation (Co-IP). According to the manufacturer's instructions (P2012, Beyotime Biotechnology, China), the centrifuged lysates were incubated with a 1:200 dilution of anti-NEK7 or anti-NLRP3 antibody overnight, at 4°C. Then, 40 µl Protein A/G Agarose (Beyotime, China) was added and incubated for an additional 4 h in a shaker. The immune complexes were boiled in the sample buffer after washing with PBS ve times. The samples were then immunoblotted with anti-NLRP3 or anti-NEK7 respectively.
Liang L., Wang G., Wang H., Bian H., & Xiao L. (2021). Oridonin relieves depressive-like behaviors by inhibiting neuroinflammation and autophagy impairment in rats subjected to chronic unpredictable mild stress.
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