Anti ddddk tag

Manufactured by Abcam

Sourced in United Kingdom

The Anti-DDDDK tag is a laboratory reagent used for the detection and purification of proteins that have been tagged with the DDDDK epitope. It is a highly specific antibody that binds to the DDDDK tag, allowing for the identification and isolation of the tagged protein from complex biological samples.

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Anti‐DDDDK tag (Binds to FLAG® tag sequence) antibody (2 citations)

3 protocols using anti ddddk tag

Flubendazole induces autophagy-mediated cell death

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Cells were purchased from American Type Culture Collection (ATCC, Manassas, VA, USA). The U937, HL60 and MCF10A cell lines were cultured in Roswell Park Memorial Institute medium (RPMI) 1640 medium and the U87, SW480, RKO, LST174t, HCT116, HepG2, HUH7, SK-HepG1, A549, 786-0, Hela, MCF-7, MDA-MB-231 and MDA-MB-468 were maintained in Dulbecco's Modified Eagle Medium (DMEM) containing 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin (Life Technologies) in 5% CO₂ at 37 °C. Cells were grown to 70-80% confluence in cell culture dishes or plates and all the experiments were performed on logarithmically growing cells.
Flubendazole (SML2510), 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (M2128), 3-MA (M9281), CQ (C6628), DAPI (D9542) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Bafilomycin A1 (ab120497) and was purchased from Abcam (Cambridge, UK). Antibodies used in this study were as follow: Beclin1 (3495, CST), SQSTM1/p62 (8025, CST), LC3B (51520, Abcam), Bax (5023, CST), Bcl-2 (2870, CST), Caspase3 (9662, CST), MMP-2 (87809, CST), E-cadherin (14472, CST), HAP1(58600, Abcam), EVE1A (216043, Abcam), HIF1α (36169, CST), β-actin (66009-1-Ig, Proteintech, IL, USA), LAMP1(25630, Abcam), ATG5 (12994, Abcam), Anti-DDDDK tag (Binds to FLAG® tag sequence) (1162, Abcam).

Zhen Y., Zhao R., Wang M., Jiang X., Gao F., Fu L., Zhang L, & Zhou X.L. (2020). Flubendazole elicits anti-cancer effects via targeting EVA1A-modulated autophagy and apoptosis in Triple-negative Breast Cancer. Theranostics, 10(18), 8080-8097.

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Comprehensive Western Blot Analysis of Key Proteins

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The specific primary antibodies involved in western blot(WB) analysis were as follows: anti- VE-cadherin (1:1000), anti-ZO-1 (1:1000), anti-TSG101(1:1000, Abcam, UK), anti-VEGFR-2 (1:1000, Abcam, UK), anti-Fas (1:1000, Abcam, UK), anti-p53 (1:1000), anti-HIPK3 (1:1000), anti-Caspase-3(1:1000, Abcam, UK), anti-HNRNPA1(1:1000), anti-HIF-1α(1:1000), anti-DDDDK tag(1:1000), anti-CD9 (1:1000, Abcam, UK), anti-CD63(1:1000, Abcam, UK), anti-ALIX(1:1000, Abcam, UK), anti-β-actin(1:1000, Proteintech, Wuhan, China), anti-SFRS2(1:1000, Abcam, UK), anti-ACO-1(1:1000, Abcam, UK). The secondary antibody was HRP-IgG (1:10000, Proteintech, Wuhan, China) and β-actin was used as internal reference.

Yang X., Wu M., Kong X., Wang Y., Hu C., Zhu D., Kong L., Qiu F, & Jiang W. (2024). Exosomal miR-3174 induced by hypoxia promotes angiogenesis and metastasis of hepatocellular carcinoma by inhibiting HIPK3. iScience, 27(2), 108955.

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Co-Immunoprecipitation of p53 and HIPK3

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Co-Immunoprecipitation (CO-IP) assay was explored to verify the interaction between p53 and HIPK3.Before this assay, HUVECs were transfected with HIPK3 overexpressing plasmids labelled with flag. Primary antibodies anti-p53(Abcam, UK) and anti-DDDDK tag (Abcam, UK) and nonspecific antibody anti-IgG (Abcam, UK) were prepared. This assay was performed according to the instructions of IP kit (Thermofisher, USA).

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