Ken Osumi, Kaoru Sato, Kensaku Murano, Haruhiko Siomi, Mikiko C. Siomi,
... siEGFP) and 0.78 (siUbc2). Immunofluorescence analysis using anti-Flag antibody to identify Flag-Ubc2 (green) expressed in ... Flag-Wde (green) co-expression. Anti-Egg and anti-Flag antibodies were used. DAPI (blue) shows the nuclei. ... upon Wde co-expression (green). Anti-Myc and anti-Flag antibodies were used. DAPI (blue) shows the nuclei. ... WT and Wde ΔCC co-expression. Anti-Myc, anti-Flag, anti-Tub, and anti-H3 antibodies were used. ... were expressed in OSCs. Immunoprecipitation was conducted using anti-Flag antibody, while anti-Myc (upper) and anti-Flag ( ...
Tópico(s): CRISPR and Genetic Engineering
2019 - Springer Nature | EMBO Reports
Jinki Yeom, Mauricio H. Pontes, Jeongjoon Choi, Eduardo A. Groisman,
... in center bottom panel of Fig 1A), and anti-FLAG antibodies pulled down CigR-HA (Fig 1A), in ... strain expressing untagged CigR (Fig 1A), and the anti-FLAG antibody did not precipitate CigR in a strain ... by immunoprecipitation and detection with anti-HA and anti-FLAG antibodies, as indicated. Relevant bands are boxed. Western ... detection was carried out with anti-MgtC and anti-FLAG antibodies, as indicated. Relevant bands are boxed. Dissociation ... detection were carried out with anti-HA and anti-FLAG antibodies. Data are representative of three independent experiments, ...
Tópico(s): Escherichia coli research studies
2018 - Springer Nature | The EMBO Journal
Xiaoqian Liu, Wen Zong, Tangliang Li, Yujun Wang, Xingzhi Xu, Zhong‐Wei Zhou, Zhao‐Qi Wang,
... into 293T cells. Immunoprecipitation (IP) was performed using anti-FLAG antibody, and immunoblotting (IB) was performed using anti-FLAG or anti-HA antibody. The experiment was repeated ... βTrCP2. IP and IB were performed using the anti-FLAG or anti-HA antibody. The experiment was repeated ... deletion mutants. IP and IB were performed using anti-FLAG or anti-HA antibody. Input in each panel ... shβTrCP1 or shβTrCP2 was examined by using an anti-FLAG antibody in 293T cells which were transfected with ... and the βTrCP2 level was monitored by an anti-FLAG antibody. β-actin was used to control the ...
Tópico(s): Genetics and Neurodevelopmental Disorders
2017 - Springer Nature | The EMBO Journal
Yaohui Chen, Li Yu, Jianfei Xue, Aihua Gong, Guanzhen Yu, Aidong Zhou, Kangyu Lin, Sicong Zhang, Nu Zhang, Cara J. Gottardi, Suyun Huang,
... its mutants were subjected to IP using mouse anti-Flag antibody, followed by IB with anti-GSK3α/β antibody (upper panel) and rabbit anti-Flag antibody (middle panel). Sequence analysis of human FoxM1 ... h. Cell lysates were subjected to IP with anti-Flag antibody, followed by IB with anti-Flag and anti-HA antibody. HA-ubiquitin was co- ... of the cells were subjected to IP with anti-Flag antibody and followed by IB with anti-FoxM1 ... was performed with lysates of the cells using anti-Flag antibody followed by IB with the indicated antibodies. ...
Tópico(s): FOXO transcription factor regulation
2016 - Springer Nature | The EMBO Journal
Roberto Marcucci, James Brindle, Simona Paro, Angela Casadio, Sophie Hempel, Nicholas A. Morrice, Andrea Bisso, Liam P. Keegan, Giannino Del Sal, Mary A. O’Connell,
... harvested, whole cell protein extracts were immunoprecipitated with anti-FLAG monoclonal antibody and analysed by immunoblot detection of ... Immunoblot analysis with anti-MPM-2 antibody of anti-FLAG immunoprecipitates from lysates of HEK293T cells transfected with ... from the neighbouring lane. ADAR input visualized with anti-FLAG antibody, lower panel. (B) Purified ADAR2 binds in ... on beads. (Upper left panel) Immunoblot analysis with anti-FLAG antibody of the binding of FLAG-tagged ADAR2, ... with GST–Pin1. Immunoblot analysis of ADAR2 with anti-FLAG antibody. Middle and lower panels are input loading ...
Tópico(s): Viral Infections and Immunology Research
2011 - Springer Nature | The EMBO Journal
Akiko Mukai, Miki Yamamoto‐Hino, Wakae Awano, Wakako Watanabe, Masayuki Komada, Satoshi Goto,
... FLAG-Fz4 was immunoprecipitated from the lysates with anti-FLAG antibody and immunoblotted with anti-Ub antibody. Fz4 ... HA-tagged Fz4, ubiquitylated Fz4 was detected by anti-FLAG immunoblotting of anti-HA immunoprecipitate (Figure 3C, left, ... expressed in COS-7 cells and immunopurified with anti-FLAG beads, were stained with Coomassie Brilliant Blue (CBB) ... with biotin and sequentially precipitated with streptavidin and anti-FLAG antibody. Immunoblotting of precipitated FLAG-Fz4K0 with anti- ... UBPY proteins were immunopurified from transfected cells using anti-FLAG antibody (Figure 3G, left) and incubated with ubiquitylated ...
Tópico(s): Developmental Biology and Gene Regulation
2010 - Springer Nature | The EMBO Journal
Holly McDonough, Peter C. Charles, Eleanor Hilliard, Shu Bing Qian, Jin-Na Min, Andrea L. Portbury, Douglas Cyr, Cam Patterson,
... with the appropriate dilutions of the primary antibodies: anti-FLAG (Sigma), anti-Myc (Santa Cruz Biotechnology), anti-Daxx ( ... min. The cell lysates were immunoprecipitated (IP) with anti-FLAG resin for immunoprecipitating Daxx, or conversely, immunoprecipitated with ... control and probed for Daxx and CHIP with anti-FLAG and anti-Myc respectively. B, COS-7 cells ... n) fractions. These fractions were then immunoprecipitated with anti-FLAG resin for Daxx and immunoblotted with anti-FLAG for Daxx and anti-Myc for CHIP. Immunoblotting ...
Tópico(s): Ubiquitin and proteasome pathways
2009 - Elsevier BV | Journal of Biological Chemistry
... MLA80 rotor for 10 min. 200 μl of anti-FLAG M2 affinity resin (Sigma) was added to 10 ... extract mix was rotated for 3 h. The anti-FLAG resins were then washed with 15 bead volumes ... 30 min and added to ∼100 μl of anti-FLAG M2 resins and IgG resins (IgG-Sepharose 6 ... the supernatant was added to 20 μl of anti-FLAG M2 resin for 2 h. The anti-FLAG M2 column was washed, and Dun1 was eluted ... B and then incubated with 3 ml of anti-FLAG M2 resin for 4 h. After washing, proteins ... described for the purification of recombinant Mrc1, except anti-FLAG resins were used during the second step of ...
Tópico(s): Microtubule and mitosis dynamics
2009 - Elsevier BV | Journal of Biological Chemistry
Akinori Endo, Naomi Kitamura, Masayuki Komada,
... μg; Zymed Laboratories Inc., South San Francisco, CA), anti-FLAG (1 μg; clone M2, Sigma), and anti-GFP ( ... 57) Google Scholar), anti-NPM (2 μg/ml), anti-FLAG (4 μg/ml), anti-GFP (4 μg/ml), ... secondary antibodies. The primary antibodies used were rabbit anti-FLAG (0.4 μg/ml), mouse anti-nucleolus (1: ... not shown). Immunofluorescence staining of transfected cells with anti-FLAG antibody showed that the mutants lacking each of ... with FLAG-tagged USP36 mutants were immunoprecipitated with anti-FLAG antibody and immunoblotted with anti-NPM (upper panel) ...
Tópico(s): Genetics and Neurodevelopmental Disorders
2009 - Elsevier BV | Journal of Biological Chemistry
Abhinav K. Jain, Shilpi Mahajan, Anil K. Jaiswal,
... immunoblotting with anti-V5HRP, anti-Tyr(P), or anti-FLAG-HRP antibodies. FLAG immunoprecipitation was done using the anti-FLAG-M2 agarose beads (Sigma). In related experiments, 1. ... binding experiment, the protein mixtures were immunoprecipitated with anti-FLAG-M2 beads (Sigma). Finally, the beads were boiled ... lysed and immunoprecipitated with either anti-V5 or anti-FLAG antibodies. The immunoprecipitate was analyzed by immunoblotting with anti-V5 and anti-FLAG antibodies. Anti-V5 antibody immunoprecipitated INrf2-V5 and ...
Tópico(s): Health, psychology, and well-being
2008 - Elsevier BV | Journal of Biological Chemistry
M Moretto Zita, Ivan Marchionni, Elisa Bottos, Massimo Righi, Giannino Del Sal, Enrico Cherubini, Paola Zacchi,
... kDa molecular weight was recognised only in the anti-FLAG immunoprecipitate obtained from CIP-untreated cell lysate (Figure ... lysate was treated with CIP and immunoprecipitated with anti-FLAG monoclonal antibody (lanes 4 and 6) and with ... on SDS-containing gels and immunoblotted using the anti-FLAG antibody. As shown in Figure 2A, only GST- ... 3) pull-down followed by Western blot with anti-FLAG antibody. (B) Endogenous gephyrin from lysates of a ... fluorescence of GFP. Pin1-FLAG was visualised by anti-FLAG antibody, followed by TRITC-conjugated secondary antibody. Scale ...
Tópico(s): Signaling Pathways in Disease
2007 - Springer Nature | The EMBO Journal
... Bjorkman, California Institute of Technology). Generation of rabbit anti-FLAG, monoclonal antibodies 3B8 2A1 against the ectodomain of ... specific to the cytoplasmic domain of TfR1, M2 anti-FLAG, mouse monoclonal anti-actin, and rabbit anti-ferritin ... and Western blot analysis and immunofluorescence with M2 anti-FLAG antibody. Transfected cells were maintained in minimal essential ... transfer to nitrocellulose, proteins were detected using M2 anti-FLAG (1:4,000), rabbit anti-TfR2 (1:10, ... secondary antibody. All mouse monoclonal primary antibodies (M2 anti-FLAG, anti-actin, 3B8 2A1, and H68.4) were ...
Tópico(s): Trace Elements in Health
2007 - Elsevier BV | Journal of Biological Chemistry
Ester Zito, Mario Buono, Stefano Pepe, Carmine Settembre, Ida Annunziata, Enrico Maria Surace, Thomas Dierks, Maria Monti, Marianna Cozzolino, Piero Pucci, Andrea Ballabio, Maria Pia Cosma,
... cellular extracts from the HL3xFS1 clone using an anti-Flag antibody (Figure 1A). These different bands of SUMF1 ... kDa were detected by Western blotting with an anti-Flag antibody. (B) SUMF1 is secreted from the HL3xFS1 ... conditioned medium. Western blotting was carried out with anti-Flag and anti--tubulin antibodies. (C) SUMF1 is secreted ... Flag-conditioned and control medium were immunoblotted with anti-Flag and anti--tubulin antibodies. (D) Endogenous SUMF1 is ... cells were analyzed by Western blotting with an anti-Flag antibody. SUMF1 was seen to be taken up ...
Tópico(s): Trypanosoma species research and implications
2007 - Springer Nature | The EMBO Journal
Isao Kuraoka, Shinsuke Ito, Tadashi Wada, Mika Hayashida, Lily Lee, Masafumi Saijo, Yoshimichi Nakatsu, Megumi Matsumoto, Tsukasa Matsunaga, Hiroshi Handa, Jun Qin, Yoshihiro Nakatani, Kiyoji Tanaka,
... HeLa cells expressing FLAG-XAB2 fusion protein using anti-FLAG antibody beads. The XAB2 complex consists of hAquarius ( ... FLAG-XAB2 complexes were purified by immunoprecipitation with anti-FLAG antibody from cell extracts. FLAG-XAB2-containing fractions ... affinity-purified using a protein A-Sepharose column. Anti-FLAG monoclonal antibody (M2) was purchased from Sigma. Anti- ... 108) Google Scholar). The extracts were incubated with anti-FLAG antibody (M2)-conjugated agarose (Sigma) and rotated at ... FLAG-XAB2 proteins were detected by immunoblotting using anti-FLAG antibody (M2). The band indicated with an asterisk ...
Tópico(s): CRISPR and Genetic Engineering
2007 - Elsevier BV | Journal of Biological Chemistry
Rei Nomura, Yoshikazu Suzuki, Akira Kakizuka, Hisato Jingami,
... GBR1-ECRs, the supernatant was applied directly to anti-FLAG M2-agarose (Sigma) packed in a disposable column ( ... and then was incubated for 1 h with anti-FLAG and anti-His antibodies at room temperature. The ... The GBR1-ECR3 protein, partially purified by the anti-FLAG M2 column, the GBR2-ECR protein, partially purified ... subjected to SDS-PAGE and Western blotting with anti-FLAG and anti-His antibodies. An anti-mouse secondary ... GBR1-ECR1–3 were expressed and purified with anti-FLAG M2-agarose gel. Eluted samples were run on ...
Tópico(s): Receptor Mechanisms and Signaling
2007 - Elsevier BV | Journal of Biological Chemistry
Shoumei Bai, Jharna Datta, Samson T. Jacob, Kalpana Ghoshal,
... PC12 cells. ZLLL (MG132), chloroquine, cycloheximide, genistein, and anti-FLAG M2 antibody were purchased from Sigma and lactacystin ... were lysed in RIPA buffer and immunoprecipitated with anti-FLAG antibody. The precipitated proteins were resolved on SDS- ... 29) Google Scholar). The membrane was reprobed with anti-FLAG M2 antibody (Sigma). In vivo labeling was done ... in RIPA buffer, and subjected to immunoprecipitation with anti-FLAG antibody. The proteins pulled down were separated by ... using PhosphorImager analysis and Western blot analysis with anti-FLAG antibody. 32P-Signal was quantified using ImageQuant software ( ...
Tópico(s): Kruppel-like factors research
2007 - Elsevier BV | Journal of Biological Chemistry
Chao Liu, Erkang Fei, Nali Jia, Hongfeng Wang, Rui-Song Tao, Atsushi Iwata, Nobuyuki Nukina, Jiang‐Ning Zhou, Guanghui Wang,
... for 1 h, the cells were incubated with anti-FLAG antibody (1:5000, Sigma) for 1 h at ... monoclonal anti-V5 antibody (1:500, Invitrogen), monoclonal anti-FLAG antibody (1:5000, Sigma), and monoclonal anti-α- ... immunofluorescence. Immunoblotting analysis revealed the smear bands of anti-FLAG immunoreactivity that represented the Ub-conjugates. In Nonidet ... excitation of EGFP and immunochemistry analysis using the anti-FLAG antibody. As shown in Fig. 2, C-K, ... FLAG-Ub was visualized by immunocytochemistry analysis using anti-FLAG anti-body. There are different patterns of α- ...
Tópico(s): Nuclear Receptors and Signaling
2007 - Elsevier BV | Journal of Biological Chemistry
Céline Tréand, Isaure du Chéné, Vanessa Brès, Rosemary Kiernan, Richard Bénarous, Monsef Benkirane, Stéphane Emiliani,
... Flag epitope to perform Tat immunoprecipitations using an anti-Flag antibody. As expected, the endogenous P-TEFb complex ... 2) were subjected to immunoprecipitation (IP) with an anti-Flag antibody (lanes 3 and 4). The immunoprecipitated material ... vectors. Lysates were subjected to immunoprecipitation using either anti-Flag or anti-HA antibodies. Immunoprecipitates were resolved by Western blot analysis using anti-Flag or anti-HA as indicated. (C) SNF5 interacts ... SNF5 plasmids were subjected to immunoprecipitation using either anti-Flag or anti-HA antibodies. Retained proteins were separated ...
Tópico(s): Genomics and Chromatin Dynamics
2006 - Springer Nature | The EMBO Journal
Sharan Paul, Warunee Dansithong, Dongho Kim, John J. Rossi, Nicholas J. G. Webster, Lucio Comai, Sita Reddy,
... MBNL2, hnRNP H and CUG-BP1 using the anti-Flag, anti-hnRNP H and anti-CUG-BP1 antibodies, ... H were measured using anti-hnRNP H or anti-Flag antibodies. (B) Normal myoblasts were first transfected with ... using anti-hnRNP H, anti-CUG-BP1 and anti-Flag antibodies are shown. The blots were re-probed ... BP1 and GFP were first incubated with the anti-Flag antibody beads. Subsequently, the immunoprecipitates were divided into ... CUG-BP1 was confirmed by immunostaining with the anti-Flag monoclonal antibody (mab) (Figure 4A). In parallel, the ...
Tópico(s): RNA Research and Splicing
2006 - Springer Nature | The EMBO Journal
Yasuhiko Kizuka, Takahiro Matsui, Hiromu Takematsu, Yasunori Kozutsumi, Toshisuke Kawasaki, Shogo Oka,
... purchased from the American Type Culture Collection. Mouse anti-FLAG M2 mAb and rabbit anti-FLAG polyclonal antibodies (pAb) were purchased from Sigma. Mouse ... After centrifugation, the clarified lysate was incubated with anti-FLAG rabbit pAb (Sigma) or anti-EGFP rabbit pAb ( ... bovine serum albumin. The cells were incubated with anti-FLAG M2 mouse mAb (Sigma) followed by incubation with ... by SDS-PAGE and Western blot analysis with anti-FLAG M2 mAb. Glucuronyltransferase activity toward asialo-orosomucoid (ASOR) ...
Tópico(s): Proteoglycans and glycosaminoglycans research
2006 - Elsevier BV | Journal of Biological Chemistry
Corinne Bousquet, Julie Guillermet‐Guibert, Nathalie Saint‐Laurent, Élodie Archer-Lahlou, Frédéric Lopez, Marjorie Fanjul, Audrey Ferrand, Daniel Fourmy, Carole Pichereaux, Bernard Monsarrat, Lucien Pradayrol, Jean‐Pierre Estève, Christiane Susini,
... line) with the cell extracts. When indicated, the anti-flag antibody was injected. Results are expressed as a ... sst2–p85 complex was assessed by using the anti-flag antibody (left panel, upper). The total input of ... down assay by immunoprecipitation/Western blot using the anti-flag antibody (right panel). Results presented in Figure 1 ... right panels, respectively, plain lines), after injecting the anti-flag antibody. This anti-flag antibody-mediated response, observed in the presence of ... B). (A, B) Immunoprecipitations were performed with the anti-flag (A) or anti-T7 (B) antibody, or preimmune ( ...
Tópico(s): Lung Cancer Research Studies
2006 - Springer Nature | The EMBO Journal
Masaaki Kawano, Takamasa Inoue, Hiroko Tsukamoto, Tatsuya Takaya, Teruya Enomoto, Ryou‐u Takahashi, Naoki Yokoyama, Noriaki Yamamoto, Akira Nakanishi, Takeshi Imai, Tadashi Wada, Kohsuke Kataoka, Hiroshi Handa,
... Tamai, MBL, Nagoya, Japan) and a mouse monoclonal anti-FLAG M2 antibody (Sigma) using the ECL detection system ( ... ng or 4.9 μg of mouse monoclonal anti-FLAG M2 antibody (Sigma). The samples were loaded onto ... 75) Google Scholar). VP1, HF-VP2, and the anti-FLAG antibody were detected by immunoblotting with anti-VP1, anti-FLAG, and anti-mouse IgG antibodies, respectively. In Vitro ... membrane, and detected by Western analysis using mouse anti-FLAG antibody as the primary antibody followed by ECL ... followed by Western analysis using anti-VP1 and anti-FLAG antibodies. VP1 alone remained in the top fraction ( ...
Tópico(s): Plant Virus Research Studies
2006 - Elsevier BV | Journal of Biological Chemistry
Ryo Yonashiro, Satoshi Ishido, Shinkou Kyo, Toshifumi Fukuda, Eiji Gotō, Yohei Matsuki, Mari Ohmura‐Hoshino, Kiyonao Sada, Hak Hotta, Hirohei Yamamura, Ryoko Inatome, Shigeru Yanagi,
... and cell lysates were immunoblotted with anti-MITOL, anti-FLAG or anti-tubulin antibodies. (B) No degradation of ... μg/ml) for 3 h and immunoblotted with anti-FLAG or anti-tubulin antibodies. (C) Polyubiquitination of MITOL ... HA-tagged ubiquitin (HA-Ub) were immunoprecipitated with anti-FLAG antibody agarose beads and immunoblotted with anti-HA or anti-FLAG antibodies. (D) Mitochondrial aggregation by MITOL CS mutant. ... exhibited autoubiquitination activity. Immunoprecipitation of FLAG-MITOL with anti-FLAG antibody from HeLa cells coexpressing FLAG-MITOL and ...
Tópico(s): RNA modifications and cancer
2006 - Springer Nature | The EMBO Journal
Jishu Wang, Reiko Shinkura, Masamichi Muramatsu, Hitoshi Nagaoka, Kazuo Kinoshita, Tasuku Honjo,
... with antibodies (Sigma, anti-c-Myc-agarose and anti-FLAG M2 affinity gel). After washing with the lysis ... their molecular sizes. Cell lysates were immunoprecipitated with anti-FLAG monoclonal antibody (mAb)-conjugated agarose (M2 beads) and ... of two AID monomers was precipitated by the anti-FLAG mAb indicating that the monomers in the AID ... immunoprecipitation was performed separately or tandemly by the anti-FLAG and anti-Myc mAbs. Obviously dialysis could not ... partners because one-step precipitation with either the anti-FLAG or anti-Myc mAb caused a strong bias ...
Tópico(s): Cytomegalovirus and herpesvirus research
2006 - Elsevier BV | Journal of Biological Chemistry
Charles A.S. Banks, Stephanie E. Kong, Henrik Spåhr, Laurence Florens, Skylar Martin‐Brown, Michael P. Washburn, Joan Conaway, Arcady Mushegian, Ronald Conaway,
... Recombinant RNasin ribonuclease inhibitor was obtained from Promega. Anti-FLAG (M2) monoclonal antibodies, anti-Myc (C-3956) rabbit polyclonal antibodies, anti-FLAG (M2)-agarose, and anti-FLAG peptide were purchased from Sigma. Light chain-specific ... proteins were purified from Sf21 cell lysates by anti-FLAG agarose immunoaffinity chromatography. Lysates from 1 × 108 cells were incubated with 0.5 ml anti-FLAG (M2)-agarose beads overnight at 4 °C. The ... 0.3 mg/ml FLAG peptide. Where indicated, anti-FLAG-agarose eluates prepared from Sf21 cells expressing both ...
Tópico(s): RNA Research and Splicing
2006 - Elsevier BV | Journal of Biological Chemistry
Takashi Ishii, M Funakoshi, Hideki Kobayashi,
... lane 4) and the extracts were immunoprecipitated with anti-FLAG beads. FLAG-Pth2 expressing at endogenous levels or ... own promoter in pth2Δ, and was immunoprecipitated with anti-FLAG M2 beads. The extracts (top and middle panels) ... al, 2002) was immunoprecipitated from yeast cells with anti-FLAG beads and then incubated with the UBL domain ... by immunoblotting of the precipitates with anti-Rpt3, anti-FLAG, anti-T7 and anti-GST. (B) Effect of ... FLAG-His6-tagged RPT1. Extracts were precipitated with anti-FLAG M2 beads, followed by immunoblotting with anti-Rpt3, ...
Tópico(s): Fungal and yeast genetics research
2006 - Springer Nature | The EMBO Journal
Ling Zhong, Ying Tan, An Zhou, Qingming Yu, Jianhua Zhou,
... Applied Science. Anti-tubulin antibody is from Oncogene. Anti-FLAG M2 antibody and fluorescence-conjugated anti-Myc and anti-FLAG antibodies are from Sigma. Anti-Myc 9E10 is ... by using anti-Myc 9E10, anti-HA 12CA5, anti-FLAG M2, or anti-parkin antibodies. For co-immunoprecipitation, ... CDCrel-1 was analyzed by Western blot using anti-FLAG M2 antibody. (b) Cells were labeled with [35S] ... was immunoprecipitated by using anti-Myc 9E10 or anti-FLAG M2 antibodies. Precipitated proteins were separated on SDS- ... days, fixed, and stained with anti-Myc and anti-FLAG antibodies or anti-parkin antibody. The localization of ...
Tópico(s): Cellular transport and secretion
2005 - Elsevier BV | Journal of Biological Chemistry
Masashi Idogawa, Tesshi Yamada, Kazufumi Honda, Satoshi Sato, Kohzoh Imai, Setsuo Hirohashi,
... truncated forms of FLAG-TCF-4 purified on anti-FLAG affinity gel were incubated with the biotinylated PARP- ... were thoroughly washed and analyzed by blotting with anti-FLAG antibody (FLAG) and avidin/horseradish peroxidase (Avidin). Full- ... length FLAG-TCF-4 protein was purified with anti-FLAG affinity gel and incubated with each form of ... by blotting with avidin-HRP (Avidin, output) or anti-FLAG antibody (FLAG). The full-length and truncated forms ... Co). Immunoprecipitation was performed with 50 μL of anti-FLAG M2 affinity gel (Sigma Chemical Co) or anti- ...
Tópico(s): Wnt/β-catenin signaling in development and cancer
2005 - Elsevier BV | Gastroenterology
Christopher Mazzochi, James K. Bubien, Peter R. Smith, Dale Benos,
... samples for 10 min at room temperature. The anti-FLAG M2 antibody (Stratagene) was diluted 1:100 with ... 1 h at room temperature. Specificity of the anti-FLAG M2 antibody was demonstrated using the FLAG peptide ( ... peptide, 10 μg of peptide, 1 μg of anti-FLAG M2 antibody for a minimum of 15 h ... C on a rotator with 3 μg of anti-FLAG mAb (Stratagene), 40 μl of a 50% slurry ... at room temperature. Following immunoprecipitation with the mAb anti-FLAG antibody, Western blot detection of α-rENaC was performed using anti-FLAG mAb (1:500 dilution) or a rabbit polyclonal ...
Tópico(s): Electrolyte and hormonal disorders
2005 - Elsevier BV | Journal of Biological Chemistry
Jingji Jin, Yong Cai, Tingting Yao, Aaron J. Gottschalk, Laurence Florens, Selene K. Swanson, José L. Gutiérrez, Michael K. Coleman, Jerry L. Workman, Arcady Mushegian, Michael P. Washburn, Ronald Conaway, Joan Conaway,
... culture in Joklik medium with 5% calf serum. Anti-FLAG Agarose Chromatography—Whole cell extracts were prepared from ... Beckman-Coulter). The resulting supernatant was subjected to anti-FLAG agarose chromatography. Nuclear extracts were prepared from HeLa ... Ti-45 rotor. Supernatants were then mixed with anti-FLAG (M2) agarose beads (Sigma) in a ratio of ... then purified Tip49a- and Tip49b-associating proteins by anti-FLAG agarose immunoaffinity chromatography. As a control for the ... the same procedure. As shown in Fig. 1, anti-FLAG agarose eluates from FLAG-Tip49a- and FLAG-Tip49b- ...
Tópico(s): Ubiquitin and proteasome pathways
2005 - Elsevier BV | Journal of Biological Chemistry