TargetMol's small compound library subsets (anti-virus compound library, anti-infection compound library, nature compound library, plant-sourced compound library, anti-inflammation library, and inhibitors library) were used for virtual screening. Small molecules selected from docking procedure were purchased from Topscience Co., Ltd. (Shanghai, China). Tetrandrine (TET) and efavirenz (EFV) were maintained in the Institute of Medicinal Biotechnology, Beijing, China. The purity of all tested compounds is more than 95%. The following antibodies were used: mouse anti-HIS (TA-02, ZSGB-Bio), mouse anti-FLAG (8146S, CST), rabbit anti-FLAG (B1020, Biodragon), mouse anti-NPC1 (ab134113, Abcam), mouse anti-P24 (ab9071, Abcam), rabbit anti-P24 (produced by our own lab), mouse anti-LAMP1 (ab24170, Abcam), mouse anti-β-actin monoclonal antibody (ab6276, Abcam), goat anti-rabbit secondary antibody (ZB-2301, Beijing Zhongshan jinqiao Biotechnology), and goat anti-mouse secondary antibody (ZB-2305, Beijing Zhongshan Jinqiao Biotechnology). Alexa Fluor-conjugated secondary antibodies were purchased from Life Technologies.
Zb 2301
Manufactured by Zhongshan Jinqiao Biotechnology
Sourced in China
The ZB-2301 is a compact and versatile laboratory centrifuge designed for a wide range of applications. It features a fixed-angle rotor that can accommodate various tube sizes, enabling efficient separation and preparation of samples. The ZB-2301 is a reliable and easy-to-use device for various laboratory settings.
Automatically generated - may contain errors
Lab products found in correlation
Zb 2305, by Zhongshan Jinqiao Biotechnology (2 mentions)
Ab9071, by Abcam (1 mentions)
Ab134113, by Abcam (1 mentions)
Ab24170, by Abcam (1 mentions)
Ta 02, by ZSGB-BIO (1 mentions)
Alexa fluor conjugated secondary antibody, by Thermo Fisher Scientific (1 mentions)
Ab6276, by Abcam (1 mentions)
Gadph, by Abcam (1 mentions)
Apobec3g, by Abcam (1 mentions)
Radioimmunoprecipitation assay buffer, by Bioss Antibodies (1 mentions)
Anti cxcr2, by Abcam (1 mentions)
Biorad versadoc imaging system, by Bio-Rad (1 mentions)
β actin, by Zhongshan Jinqiao Biotechnology (1 mentions)
Ab205587, by Abcam (1 mentions)
Chemiluminescence gel imaging system, by Bio-Rad (1 mentions)
Ab133615, by Abcam (1 mentions)
Ab92726, by Abcam (1 mentions)
Df6087, by Affinity Biosciences (1 mentions)
Ta 09, by Zhongshan Jinqiao Biotechnology (1 mentions)
Ab109201, by Abcam (1 mentions)
4 protocols using zb 2301
1
Antiviral Compound Screening Library
2
Protein Expression Analysis Protocol
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The protein samples (40 μg) were boiled for 5 min and then immediately placed in an ice bathed for 5 min, then centrifuged at 12 000 rpm for 5 min at 4°C. The supernatant (25 μg) were electrophoresed in sodium dodecyl sulfate polyacrylamide gel (SDS-PAGE). After electrophoresis, the targeted proteins on the gel were transferred onto a polyvinylidene difluoride (PVDF) membrane and blocked with 5% BSA for 1 h at room temperature. After washing with TBST for 5 min, the PVDF membrane was incubated with 5 ml primary antibodies overnight at 4°C. After 3 washes with TBST, the membrane was incubated with 10 mL secondary antibodies diluent for 1 h at room temperature. After 3 washes with TBST, the ECL Chemiluminescence reagent A and B (100 μl) were mixed 1: 1 and uniformly applied near the target strip. The primary antibodies used in this study were as follows: APOBEC3G (Bioss, MA, USA), HPV E6 (Abcam, Cambridge, MA), p53 (Abcam, Cambridge, MA), with GADPH (Abcam, Cambridge, MA) and was considered as the internal reference. The secondary antibodies used in this study were: ZB-2301 (Zhongshan Jinqiao Biotechnology Co., Beijing, China) and ZB-5305 (Zhongshan Jinqiao Biotechnology Co., Beijing, China). The experiments were repeated 3 times.
3
Western Blot Analysis of CXCR1 and CXCR2 in Liver Cancer Cells
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In brief, liver cancer cells were lysed using radioimmunoprecipitation assay buffer (BIOSS) for 30 min on ice. The cell lysate was centrifuged at 12,000 × g for 20 min at 4°C and the supernatant was collected. Protein concentration was determined by BCA protein assay. A total of 30 µg protein/lane was separated by SDS-PAGE on a 10% gel and transferred onto nitrocellulose filter membranes. Following blocking with 5% skim milk for 1 h at room temperature, the membranes were incubated with primary antibodies overnight at 4°C, and then with secondary antibodies for 2 h at room temperature. The primary antibodies used were as follows: Rabbit polyclonal anti-CXCR1 (1:2,000; cat. no. ab137351; Abcam) and anti-CXCR2 (1:2,000; cat. no. ab14935; Abcam) antibodies; monoclonal mouse anti-GAPDH (1:2,000; cat. no. TA-08; Zhongshan Jinqiao Biotechnology) and β-actin (1:2,000, cat. no. TA-09; Zhongshan Jinqiao Biotechnology, Beijing, China). The goat anti-mouse (cat. no. ZB2305; 1:10,000) and anti-rabbit secondary antibodies (cat. no. ZB2301; 1:10,000) were purchased from Zhongshan Jinqiao Biotechnology. The protein bands were visualized by ECL reagents (cat. no. C05-07003; BIOSS). Images were captured and the intensity of the bands was quantitated with the Bio-Rad Versa Doc imaging system (Bio-Rad Laboratories, Inc.).
4
Western Blot Analysis of Exosomes and Inflammatory Markers
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Prepared protein samples (10 μg) were subject to 12% SDS-PAGE, followed by electro-transfer onto PVDF membranes. Then the membranes were blocked in 5% skimmed milk for 1 h. Antibodies against exosome markers CD9 (ab92726; Abcam, Cambridge, UK), CD81 (ab109201; Abcam) and calnexin (ab133615; Abcam) were diluted at a ratio of 1:1000. The inflammatory response of vascular tissue and VSMCs was detected using antibodies against NFATc3 (18222-1-AP, 1:1000 dilution; Proteintech, Wuhan, China) and IL-6 (DF6087, 1:500 dilution; Affinity Biosciences, Cincinnati, USA), IL-1β (DF6251, 1:500 dilution; Affinity Biosciences) and TNF-α (ab205587, 1:500 dilution; Abcam). The internal control anti-β-actin primary antibody (TA-09; Zhongshan Jinqiao Biotechnology, Beijing, China) was used at 1:1000 dilution. The PVDF membranes were incubated with the above primary antibodies at 4°C overnight, followed by incubation with HRP-conjugated goat anti-mouse IgG (ZB-2305, 1:4000; Zhongshan Jinqiao Biotechnology) or HRP-conjugated goat anti-rabbit IgG (ZB-2301, 1:4000; Zhongshan Jinqiao Biotechnology) secondary antibodies for 1 h at room temperature. After extensive wash, protein bands were visualized using ultra high sensitivity ECL kit (HY-K1005; MCE, New Jersey, USA), and images were obtained with a chemiluminescence gel imaging system (12003153; Bio-Rad, Hercules, USA).
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