Commercial antibodies used were anti-AnkG (Santa Cruz), anti-Slack (NeuroMab), anti-NaV1.2 (Alomone), anti-NaV1.6 (Alomone), anti-HA (Abbkine), anti-Flag (Abbkine), anti-β-actin (Biodragon), HRP goat anti-mouse IgG LCS (Abbkine), HRP mouse anti-rabbit IgG LCS (Abbkine), Alexa Fluor 488-AffinityPure Fab Fragment donkey anti-rabbit IgG (Jackson), and Alexa Fluor 594 donkey anti-mouse IgG (Yeason). GPCR Extraction Reagent was from Pierce, NP40 lysis buffer was from Beyotime, protease inhibitor mixture cocktail was from Roche Applied Science, rabbit IgG and mouse IgG were from Santa Cruz, and Protein G Dynabeads were from Invitrogen. Tetrodotoxin was from Absin Bioscience. Quinidine was from Macklin, and riluzole was from Meilunbio. All other reagents were purchased from Sigma-Aldrich.
Anti flag
Manufactured by Abbkine
Sourced in China, United States
Anti-Flag is a laboratory equipment product designed for immunoprecipitation and immunoblotting applications. It functions as an antibody that specifically binds to the FLAG epitope tag, a commonly used protein tag for the identification and purification of recombinant proteins.
Automatically generated - may contain errors
Lab products found in correlation
Anti nav1, by Alomone (2 mentions)
Protease inhibitor mixture cocktail, by Roche (1 mentions)
Anti ankg, by Santa Cruz Biotechnology (1 mentions)
Anti ha, by Abbkine (1 mentions)
Quinidine, by Maclin Biochemical Technology (1 mentions)
Protein g dynabead, by Thermo Fisher Scientific (1 mentions)
Np 40 lysis buffer, by Beyotime (1 mentions)
Anti p akt, by Cell Signaling Technology (1 mentions)
Anti rhoc, by Cell Signaling Technology (1 mentions)
Tae226, by Selleck Chemicals (1 mentions)
Anti p fak, by Cell Signaling Technology (1 mentions)
Anti p pyk2, by Cell Signaling Technology (1 mentions)
Ly294002, by Selleck Chemicals (1 mentions)
U0126, by Selleck Chemicals (1 mentions)
Anti pmapk, by Cell Signaling Technology (1 mentions)
Sw620, by National Collection of Authenticated Cell Cultures (1 mentions)
Sw480, by National Collection of Authenticated Cell Cultures (1 mentions)
Hct116, by National Collection of Authenticated Cell Cultures (1 mentions)
Anti body, by Abbkine (1 mentions)
Ripa lysis buffer, by Beyotime (1 mentions)
4 protocols using anti flag
1
Investigating Sodium Channel Interactome
2
Co-immunoprecipitation of PUB23 and GT1
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The coding sequence of PUB23 was cloned into vector pD2P (Clontech, USA) in frame with FLAG tag. Coding sequence (1089 bp) of the positive interacting protein (GT1) was cloned into pD2P in frame with 6×His tag. The recombined FLAG-PUB23 and His-GT1 were expressed by cell free expression system according to the protocol handbook (Nanjing Ruiyuan Biotech, Nanjing, China) and purified with FLAG-tag magnetic beads or His-tag magnetic beads, respectively. The purified FLAG-PUB23 and His-GT1 were co-incubated and precipitated with FLAG-tag magnetic beads or His-tag magnetic beads, respectively. Components were separated on a SDS-PAGE gel and immunoblotted with anti-FLAG (Abbkine, Wuhan, China; no. A02010) or anti-His (Abbkine, China; no. D-AKE2054) antibody.
3
CRC Cell Lines Signaling Pathway Analysis
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All four CRC cell lines (LOVO, SW620, SW480 and HCT116) and the 293T cell line were purchased from Cell Bank of Chinese Academy of Sciences (Shanghai, China). The cell lines were cultured at 37 °C in a 50 mL/L CO2-humidified atmosphere with the appropriate medium according to the requirements of the Cell Bank. Anti-(p-) Pyk2 (proline-rich tyrosine kinase 2), anti-(p-) FAK, anti-(p-) MAPK (Mitogen activated protein kinase), anti-(p-) AKT and anti-RhoC antibodies were purchased from Cell Signaling Technology. Anti-flag, anti-VEGF (vascular endothelial growth factor) and anti-FMNL3 antibodies were obtained from Abbkine, Inc (Redlands, CA, United States) and Abnova (Taiwan, China), respectively. For inhibitor treatment, 1 μmol/L TAE226 (Selleck), 20 μmol/L U0126 (Selleck) or 20 μmol/L Ly294002 (Selleck) was added to the cultured cells for 48 h, respectively.
4
Immunoblotting of ZmWAX2 Overexpression
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Total proteins were extracted from the ZmWAX2 overexpression lines and the corresponding wild type using the RIPA lysis buffer (P0013C; Beyotime, Shanghai, China). Equal amounts of proteins were separated on 8% SDS‐PAGE gel and transferred to PVDF membranes. After blocking in 5% no‐fat milk, the membranes were incubated in anti‐body (1:10000, Anti‐Flag, Abbkine, A02010, Wuhan, China) at 4 °C overnight following incubation in secondary anti‐body (1:10 000, Abbkine A23210) for 2 h at room temperature. The Coomassie brilliant blue staining of the large subunit of rubisco protein was used as the loading control.
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