Mouse anti flag

Manufactured by Proteintech

Mouse anti-Flag is a monoclonal antibody that recognizes the Flag epitope tag. It is commonly used in research applications for the detection and immunoprecipitation of Flag-tagged proteins.

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Close spelling variants of this product

Anti-Flag (86 citations) Mouse monoclonal anti-Flag antibodies (6 citations) Mouse anti-Flag antibody (5 citations) Mouse anti-Flag tag (4 citations) Anti-TM (2 citations) Mouse anti-flag tag monoclonal antibody (2 citations) Mouse anti-FLAG IgG (2 citations)

3 protocols using mouse anti flag

Antibody and Chemical Reagents for PRRSV Study

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Primary antibodies were mouse anti-PRRSV N protein (produced in our laboratory), mouse anti-β-actin (Proteintech, #A5316), mouse anti-Flag (Proteintech, #80010-1-RR), rabbit anti-LC3 (Proteintech, #14600-1-AP), rabbit anti-ATG7 (Cell Signaling Technology, #2631), rabbit anti-CaMKII (Abcam, #ab168818), rabbit anti-p-AMPK (Thr172) (Cell Signaling Technology, #2535), rabbit anti-AMPK (Cell Signaling Technology, #2532), rabbit anti-p-mTOR(Ser2448) (Cell Signaling Technology, #5536), rabbit anti-mTOR (Cell Signaling Technology, #2983), rabbit anti-STIM1 (Cell Signaling Technology, #4916), mouse anti-Orai1 (Proteintech, #66223–1), rabbit anti-GRP78 (Proteintech, #11587-1-AP) and rabbit anti-Calnexin (Abcam, #ab92573). HRP-labeled rabbit or mouse secondary antibodies were purchased from Beyotime (China).
Chemicals used included 2-Aminoethyl diphenylborinate (2-APB) (Selleck, #S6657), 3-MA (Selleck, #S2767), 4-Phenylbutyric acid (4-PBA) (Selleck, #S3592), BAPTA-AM (Selleck, #S7534), Dantrolene sodium (Selleck, #S5478), Dorsomorphin (Compound C) 2HCL (Selleck, #S7306), KN-93 (Selleck, #S7423), ML-9 HCL (Selleck, #S6847), Procaine (Selleck, #S4668), Rapamycin (MedChemExpress, #HY-10219), Tetracaine-HCL (Selleck, #S2573), Thapsigargin (Selleck, #S7895), Tunicamycin (Beyotime, #SC0393).

Diao F., Jiang C., Sun Y., Gao Y., Bai J., Nauwynck H., Wang X., Yang Y., Jiang P, & Liu X. (2023). Porcine reproductive and respiratory syndrome virus infection triggers autophagy via ER stress-induced calcium signaling to facilitate virus replication. PLOS Pathogens, 19(3), e1011295.

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Western Blot Analysis of Subcellular Proteins

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For Western blot analysis, cells were harvested and lysed in RIPA lysis buffer (Cell Signal Technology). To investigate subcellular distribution of proteins, nuclear and cytoplasmic fractions were enriched using Nuclear and Cytoplasmic Protein Extraction Kit (CoWin Bioscience). Western blot assays were followed according to an established protocol (20 (link)). Antibody used included mouse anti-GFP (Abmart), rabbit anti-acetyl Lysine (Abcam), rabbit anti-SUMO1 (Abcam), rabbit anti-Lamin B1 (Abcam), mouse anti-Ub (Santa Cruz), rabbit anti-USP49 (NOVUS), mouse anti-Flag (Proteintech), mouse anti-GAPDH (Thermo Fisher Scientific), mouse anti–β-tubulin (Thermo Fisher Scientific), rabbit anti-GRα (Abcam), rabbit anti-GRβ (Abcam). For protein stability detections, cells were treated with proteasome inhibitor MG132 (20 μmol/L, Sigma-Aldrich) for 6 hours or protein synthesis inhibitor Cycloheximide (CHX, 100 μg/mL, Sigma-Aldrich) for indicate time before harvest.

Hu Y., Jiang Y., Zhang Z., Wang J., Zhang B., Gong L., Ji L., Pu Z., Yang X., Zou J, & Yin Y. (2021). Oncogenic Activity of Glucocorticoid Receptor β Is Controlled by Ubiquitination-Dependent Interaction with USP49 in Glioblastoma Cells. Molecular Cancer Research, 20(1), 92-101.

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Co-Immunoprecipitation Analysis of Protein Interactions

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Cells were lysed in RIPA buffer with 1 mM PMSF (Solarbio), then centrifuged at 4°C and 12,000 rpm to pellet the debris. The supernatants were boiled in SDS sample buffer, resolved on 10% SDS‐PAGE gels and transferred onto 0.22‐μm PVDF membranes (Millipore). Co‐IP was performed according to the manufacturer's procedure (Absin). The following antibodies were used: rabbit anti‐TRA2A (1:1000, 72625, Abcam), rabbit anti‐GADPH (1:1000, 2118, CST), rabbit anti‐RBMX (1:1000, 14794, CST), rabbit anti‐KIAA1429 (1:1000, 88358, CST), rabbit anti‐Flag (1:2000, 20543, Proteintech), mouse anti‐Flag (1:2000, 66008, Proteintech), rabbit anti‐EZH2 (1:4000, 5246, CST), rabbit anti‐β‐catenin (1:1000, 8480, CST), anti‐rabbit IgG‐HRP (1:1000, 7074, CST), and anti‐mouse IgG‐HRP (1:2000, 7076, CST).

Bei M., Hao S., Lin K., Chen Q., Cai Y., Zhao X., Jiang L., Lin L., Dong G, & Xu J. (2023). Splicing factor TRA2A contributes to esophageal cancer progression via a noncanonical role in lncRNA m6A methylation. Cancer Science, 114(8), 3216-3229.

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