Gfp tag

Manufactured by ABclonal

Sourced in United States

The GFP tag is a protein tag derived from the green fluorescent protein (GFP) found in the jellyfish Aequorea victoria. It is commonly used as a reporter or marker in molecular biology and cell biology experiments to study protein localization, expression, and interactions within living cells.

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Lab products found in correlation

A19607, by ABclonal (1 mentions) β tubulin, by Merck Group (1 mentions) N cadherin, by Cell Signaling Technology (1 mentions) P0013, by Beyotime (1 mentions) Ab236632, by Abcam (1 mentions) A2348, by ABclonal (1 mentions) β actin, by ABclonal (1 mentions) Ab222320, by Abcam (1 mentions) Cell lysis buffer for western and ip, by Beyotime (1 mentions) Ha tag, by Abcam (1 mentions) Hp mlkl, by Abcam (1 mentions) Sting, by Proteintech (1 mentions) Myc tag, by ABclonal (1 mentions) Flag tag (flag), by ABclonal (1 mentions) Gapdh, by Proteintech (1 mentions) β tubulin, by ABclonal (1 mentions) β tubulin, by Developmental Studies Hybridoma Bank (1 mentions)

4 protocols using gfp tag

Western Blot Analysis of Cellular Proteins

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After being washed with Phosphate Buffered Saline (PBS), cells were lysed by a protein lysis buffer (P0013, Beyotime) containing 1 mM of Phenylmethanesulfonyl fluoride (PMSF) (ST506, Beyotime (Shanghai, China)). Protein lysates were quantified by Bradford assay (1863028, Thermo Fisher Scientific (Waltham, Massachusetts, USA)). Western blotting was carried out according to standard methods. Antibodies specific to fibronectin (1561301-AP, Proteintech (Wuhan, Hubei Province, China)), CK19 (10712-AP, Proteintech), CK7 (15539-1, Proteintech), CK20 (17329-1-AP, Proteintech), N-cadherin (13116S, Cell Signaling Technology), β-tubulin (T8328, Sigma-Aldrich), MRP3 (39909S, Cell Signaling Technology (Danvers, Massachusetts, USA)), Flag tag (F1804, Signal-Aldrich (Burlington, Massachusetts, USA)), GFP tag (AED011, ABclonal (Wuhan, Hubei Province, China)),Vimentin (A19607, ABclonal), LT-SV40 (sc-147, Santacruz (Dallas, Texas, USA)), hTERT (17329-1-AP, Proteintech), TP53 (2527S, CST), and Rb (9313S, CST) were applied for Western blotting.

Wang Z., Wang S., Jia Z., Zhao Y., Yang M., Yan W., Chen T., Xiang D., Shao R, & Liu Y. (2022). Establishment and characterization of an immortalized epithelial cell line from human gallbladder. Frontiers in Oncology, 12, 994087.

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Western Blot Analysis of Cellular Proteins

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The treated cells were lysed using RIPA buffer containing protease and phosphatase inhibitors. Equal amounts of protein samples were separated on SDS-PAGE and transferred onto nitrocellulose membranes (Pall, Port Washington, USA). Next, the membranes were blocked with 5% nonfat milk for 1 h at room temperature. Then the membranes were incubated with primary antibodies at 4 °C overnight and incubated with secondary antibody for 1 h at room temperature. The information of antibodies were as follow: DPT (ab255823, Abcam, UK), Yes-associated protein 1 (YAP1, A1002, ABclonal, China), Phospho-YAP (Ser127) (#13008, CST, USA), GAPDH (#5174, CST, USA), β-actin (AC026, ABclonal, China), Histone H3 (A2348, ABclonal, China), HA tag (ab236632, Abcam, UK), DYKDDDDK Tag (#14793, CST, USA), and GFP-Tag (AE011, ABclonal, China).

Ye D., Wang Y., Deng X., Zhou X., Liu D., Zhou B., Zheng W., Wang X, & Fang L. (2023). DNMT3a-dermatopontin axis suppresses breast cancer malignancy via inactivating YAP. Cell Death & Disease, 14(2), 106.

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Immunoprecipitation and Western Blot Analysis

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Cells were treated with cell lysis buffer for Western and IP (Beyotime). The indicated antibodies were added to cell lysates or supernatant, followed by incubation with gentle rocking overnight at 4°C. Then, protein G agarose beads were added to cell lysates, which was incubated with gentle rocking for 4 h at 4°C. Next, samples were eluted in SDS‐PAGE sample loading buffer and boiled. For immunoprecipitation, anti‐FLAG (AE005; ABclonal), STING (19851‐1‐AP; Proteintech) and MLKL (PA5‐102810; Invitrogen) antibodies were used.
The primary antibodies were used including STING (19851‐1‐AP; Proteintech), hp‐STING (S366, 50907; CST), mp‐STING (S365, 72971; CST), p‐TBK1 (S172; CST), TBK1 (38066; CST), IRF3 (4302; Cell Signaling), p‐IRF3 (S396, 29047; CST), mp‐RIPK3 (T231+S232, ab222320; Abcam), mp‐MLKL(S345, ab196436; Abcam), hp‐RIPK3 (S227, ab209384; Abcam), hp‐MLKL (S358, ab187091; Abcam), hMLKL (A19685; ABclonal), mMLKL (37705; CST), hRIPK3 (86671; CST), mRIPK3 (15828; CST), mp‐RIPK1 (S166, 31122; CST), RIPK1 (3493; CST), LC3 (12741; CST), P62 (A7758; ABclonal), β‐actin (4970; CST), β‐Tubulin (AC021; ABclonal), GAPDH (60004‐1‐Ig; Proteintech), FLAG‐tag (AE063; ABclonal), HA‐tag (ab236632; Abcam), MYC‐tag (AE070; ABclonal) and GFP‐tag (AE012; ABclonal).

Zhang X., Wu J., Liu Q., Li X., Yang Y., Wu L., Wu X., Zhao Y, & Ren J. (2023). RIPK3–MLKL necroptotic signalling amplifies STING pathway and exacerbates lethal sepsis. Clinical and Translational Medicine, 13(7), e1334.

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Antibody Characterization for Drosophila Studies

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The antibodies used in this study are listed in KEY RESOURCES TABLE. Note that the primary antibodies against PLIN2 (PROGEN, Cat# GP40S) and PLIN3 (PROGEN, Cat# GP30S) showed cross-species reactivity to human and mouse orthologs, and the antibodies against β-Actin (ABclonal, Cat# AC026) and β-Tubulin (Developmental Studies Hybridoma Bank, Cat# E7) showed cross-species reactivity to human, mouse and fly orthologs. Three rabbit polyclonal antibodies generated in this study are listed as follows: A rabbit polyclonal antibody was raised against full-length Drosophila Plin2 (ABclonal). In immunoblotting, a doublet of bands with apparent molecular weights of 44 kDa and 46 kDa were observed, corresponding to two endogenous protein products of plin2 transcripts with alternatively spliced exons 3. The antibody specificity was confirmed by immunoblotting (1:2000) and immunofluorescence (1:200) (Figures S1A andS1B). A rabbit polyclonal antibody was raised against N-terminal 307 amino acids of Drosophila Ubr1 (ABclonal). The Antibody specificity was confirmed by immunoblotting (1:2000) (Figure 2C). A rabbit polyclonal antibody was raised against GFP tag (ABclonal). Its specificity was confirmed by immunoblotting (1:3000).

Zhang Y., Lin S., Peng J., Liang X., Yang Q., Bai X., Li Y., Li J., Dong W., Wang Y., Huang Y., Pei Y., Guo J., Zhao W., Zhang Z., Liu M, & Zhu A.J. (2022). Amelioration of hepatic steatosis by dietary essential amino acid-induced ubiquitination. Molecular cell, 82(8).

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