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Rabbit anti flag

Manufactured by Proteintech
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Rabbit anti-FLAG is a primary antibody that specifically recognizes the FLAG epitope tag. The FLAG tag is a widely used, short peptide tag that can be added to recombinant proteins to facilitate their detection and purification. Rabbit anti-FLAG antibody can be used to detect and immunoprecipitate FLAG-tagged proteins in various applications such as Western blotting, immunoprecipitation, and immunocytochemistry.

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Lab products found in correlation

Mouse anti flag, by Merck Group (4 mentions) Anti gapdh, by Huabio (1 mentions) Anti phospho ikkα β, by Cell Signaling Technology (1 mentions) Hek293t cell, by American Type Culture Collection (1 mentions) Anti phospho iκbα, by Cell Signaling Technology (1 mentions) Human recombinant tnfα, by R&D Systems (1 mentions) Anti iκbα, by Santa Cruz Biotechnology (1 mentions) Lipo293 transfection reagent, by Beyotime (1 mentions) Dual specific luciferase assay kit, by Promega (1 mentions) Anti lmnb1, by Proteintech (1 mentions) Anti ha, by OriGene (1 mentions) Anti tnf α, by Proteintech (1 mentions) Anti ikk β, by Proteintech (1 mentions) Anti p65, by Santa Cruz Biotechnology (1 mentions) Anti phospho nf κb p65, by Cell Signaling Technology (1 mentions) Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions) Anti β actin, by Cell Signaling Technology (1 mentions) Normal rabbit igg, by ABclonal (1 mentions) Rabbit anti ev a71 vp1, by GeneTex (1 mentions) Siphb2, by RiboBio (1 mentions)

Close spelling variants of this product

Rabbit anti-Flag polyclonal antibody Rabbit anti-FLAG antibody Anti-Flag Anti-TM

9 protocols using rabbit anti flag

1

Characterization of Cellular Response to TNF-α

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HEK293T cells (ATCC, CRL-3216™). A549 cells (SCSP-503) and THP-1 cells (TCHu 57) were purchased from the National Collection of Authenticated Cell Cultures (Shanghai, China). Sendai virus (SeV) was kindly provided by Prof. Yanyi Wang (Wuhan Institute of Virology, CAS) and propagated in SPF chicken embryonated eggs. Lipo293™ Transfection Reagent (Beyotime Biotechnology, C0521), Lipofectamine 2000 (Invitrogen, 11,668,019), dual-specific luciferase assay kits (Promega, E1980), human recombinant TNFα (R&D Systems, 210-TA-020/CF), BAY 11–7082 (MCE, HY-13,453), mouse anti-Flag (Sigma, F3165), rabbit anti-Flag (Proteintech, 20,543–1-AP), anti-β-actin (Cell Signaling Technology, # 3700S), anti-GAPDH (HuaBio, #R1210–1), anti-LMNB1(Proteintech, 2987–1-AP), anti-HA (Origene, TA100012), anti-TNF-α (Proteintech, 60,291–1-Ig), anti-phospho-NF-κB p65(Cell Signaling Technology, #3033S), anti-phospho-IκBα (Cell Signaling Technology, #9246S), anti-phospho-IKKα/β (Cell Signaling Technology, # 2697S), anti-IKK-β (Proteintech, 15,649–1-AP) anti-IκBα (Santa Cruz, sc-1643), anti-p65 (Santa Cruz, sc-8008), donkey anti-mouse IgG-Cy3 (Absin, abs20015) and goat anti-Rabbit IgG-FITC (Absin, abs20004ss) were purchased from the indicated companies.
Nie Y., Mou L., Long Q., Deng D., Hu R., Cheng J, & Wu J. (2023). SARS-CoV-2 ORF3a positively regulates NF-κB activity by enhancing IKKβ-NEMO interaction. Virus Research, 328, 199086.
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2

Investigating Autophagy Regulation via PHB2

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The following antibodies were used: rabbit anti-LC3B (Cell Signaling Technology, Boston, MA, USA, 3868), rabbit anti-PHB2 (Cell Signaling Technology, 14085), rabbit anti-SQSTM1/p62 (ABclonal Technology, Wuhan, China, A77580), normal rabbit IgG (ABclonal Technology, AC005), mouse anti-PHB2 (Proteintech, Wuhan, China, 66424-1-Ig), mouse anti-β-actin (Proteintech, 66009-1-Ig), rabbit anti-flag (Proteintech, 20543-1-AP), and rabbit anti-EV-A71 VP1 (Genetex, Irvine, CA, USA, GTX132339).
The following reagents were used: 3-methyladenine (3-MA; Selleck, Shanghai, China, S2767), rapamycin (Selleck, S1039), nontargeting siRNA (SiNC) and siRNA targeting PHB2 (SiPHB2) and ATP6AP2 (SiATP6AP2) (RiboBio, Guangzhou, China), Lipofectamine 3000 Reagent (Invitrogen, Carlsbad, CA, USA, L3000015), Lipofectamine RNAiMAX Reagent (Invitrogen, 13778-150), and primers for PCR and real-time quantitative PCR (RT-qPCR) (Invitrogen).
Su W., Huang S., Zhu H., Zhang B, & Wu X. (2020). Interaction between PHB2 and Enterovirus A71 VP1 Induces Autophagy and Affects EV-A71 Infection. Viruses, 12(4), 414.
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3

Immunoblotting with Diverse Antibodies

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The following antibodies were used for immunoblotting: Rat anti-HA (Roche, ROAHAHA), Rabbit anti- FLAG (Proteintech, 20543-1-AP), Mouse anti-GAPDH (Developmental Studies Hybridoma Bank, hGAPDH-2G7), Rabbit anti-USP46 (Proteintech, 13502-1-AP), Mouse anti-Tubulin (Developmental Studies Hybridoma Bank, E7), Mouse anti-β-catenin (BD Transduction Laboratory, 610154), Rabbit anti-Axin1 (Cell Signaling Technology, 2087), Rabbit anti-Insulin Receptor β (Cell Signaling Technology, 3025), Rabbit anti-LRP6 (Cell Signaling Technology, 2560), Rabbit anti-WDR20 (Bethyl Laboratories, A301-657A), Rabbit anti-WDR48 (UAF1) (Proteintech, 16503-1-AP), Goat anti-rat IgG H + L-HRP (Thermo, 31470), Goat anti-mouse IgG H + L-HRP (Promega, W4021), Goat anti-rabbit IgG H + L-HRP (Promega, W4011). All primary antibodies were used at 1:1000 dilution except anti-FLAG (1:2000), anti-GAPDH (1:500), and anti-WDR20 (1:2500). All secondary HRP antibodies were used at 1:5000 dilution.
Ng V.H., Spencer Z., Neitzel L.R., Nayak A., Loberg M.A., Shen C., Kassel S.N., Kroh H.K., An Z., Anthony C.C., Bryant J.M., Lawson A., Goldsmith L., Benchabane H., Hansen A.G., Li J., D’Souza S., Lebensohn A.M., Rohatgi R., Weiss W.A., Weiss V.L., Williams C., Hong C.C., Robbins D.J., Ahmed Y, & Lee E. (2023). The USP46 complex deubiquitylates LRP6 to promote Wnt/β-catenin signaling. Nature Communications, 14, 6173.
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4

Comprehensive Antibody Panel for Cellular Adhesion

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Antibodies used for western blot analysis: mouse anti-GFP (1:1000; Proteintech, #50430-2-AP), rabbit anti-pFAK Y576 (1:200; Santa Cruz, #sc-16563), rabbit anti-pFAK Y397 (1:200; Novus, #NBPI-60837), rabbit anti-pPaxillin Y31 (1:200; Santa Cruz, #sc-14035), mouse anti-FAK (1:1000; Proteintech, #66258-I-1g), mouse anti-Paxillin (1:5000; BD Biosciences, #610051), rabbit anti-FLAG (1:1000; Proteintech, #80010-1-RR).
Antibodies used in immunofluorescence experiments: mouse anti-FAK 4.47 (1:1000; Millipore, #05-537), mouse anti-FAK (1:1000; Proteintech, #66258-I-1g), rabbit anti-FAK C-20 (1:200; Santa Cruz, #sc-558), rabbit anti-pFAK Y576 (1:200; Santa Cruz, #sc-16563), rabbit anti-pFAK Y397 (1:200; Novus, #NBPI-60837) goat anti-Talin C-20 (1:750; Santa Cruz, #sc-7534), mouse anti-Paxillin (1:750; R&D Systems, #AF4259), mouse anti-Vinculin (1:1000; Sigma, #V9131), rabbit anti-Vinculin (1:2000; Proteintech, #26520-I-AP), mouse anti-av Integrin H-2 (1:100; Santa Cruz, #sc-376156), rabbit anti-pPaxillin Y31 (1:200; Santa Cruz, #sc-14035), mouse anti-p-Tyr (pY20) (1:200; Santa Cruz, #sc-508), rat anti-active β1-Integrin 9EG7 (1:700; BD Bioscences, #550531), mouse anti-p130 Cas 35B.1A4 (1:500; Santa Cruz, #sc-20029), mouse anti-Tensin C-2 (1:1000; Santa Cruz, #sc-376367).
Antoniades I., Kyriakou M., Charalambous A., Kalalidou K., Christodoulou A., Christoforou M, & Skourides P.A. (2021). FAK displacement from focal adhesions: a promising strategy to target processes implicated in cancer progression and metastasis. Cell Communication and Signaling : CCS, 19, 3.
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5

Antibody Characterization for Western Blot and Immunostaining

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The following primary antibodies were used for Western blotting and immunostaining in this study: mouse anti-Flag (Sigma-Aldrich, F3165), rabbit anti-Flag (Proteintech, 20543-1-AP), rabbit anti-Vps4B (Proteintech, 17673-1-AP), mouse anti-P62 (Abcam, ab56416), rabbit anti-LC3B (Abcam, ab48394), mouse anti-V5 (Proteintech, 66007-1-Ig), mouse anti–glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (Proteintech, 60004-1-Ig), mouse anti–β-tubulin class III (Tuj 1; Abcam, ab78078), rabbit anti–Tuj 1 (Sigma-Aldrich, T2200), and mouse anti–β-actin (Cell Signaling Technology, 3700). The horseradish peroxidase–conjugated secondary antibodies were as follows: goat anti-mouse (Sigma-Aldrich, A4416) and goat anti-rabbit (Sigma-Aldrich, A9169). The fluorophore-conjugated secondary antibodies were as follows: goat anti-rabbit Alexa Fluor 488 (Life Technologies, A11036), goat anti-mouse Alexa Fluor 488 (Life Technologies, A11029), goat anti-mouse Alexa Fluor 568 (Life Technologies, A11031), goat anti-rabbit Cy5 (Life Technologies, A10523), and goat anti-mouse Cy5 (Life Technologies, A105234).
Wang H., Wang X., Zhang K., Wang Q., Cao X., Wang Z., Zhang S., Li A., Liu K, & Fang Y. (2019). Rapid depletion of ESCRT protein Vps4 underlies injury-induced autophagic impediment and Wallerian degeneration. Science Advances, 5(2), eaav4971.
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6

Antibody Verification for Protein Studies

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Goat anti-FE65 (E-20), mouse anti-ARF6 (3A-1), mouse anti-ARNO (H-7), mouse anti-c-Jun (D-11) and mouse anti-α-Tubulin (DM1A) were purchased from Santa Cruz. Rabbit anti-ARNO, rabbit anti-GFP, rabbit anti-His and rabbit anti-FLAG were obtained from Proteintech. Mouse anti-myc antibody (9B11) and rabbit anti-COX IV (3E11) were obtained from Cell Signaling Technology. Mouse anti-pan-cadherin (C1821), mouse anti-β-COP (maD) and mouse anti-FLAG antibody (M2) were obtained from Sigma. Goat anti-GST antibody was obtained from GeneTex. Mouse anti-GAPDH (AM4300) was purchased from Ambion. Rabbit anti-β-Tubulin was purchased from Abcam. Rabbit anti-FE65 was as previously described [13 (link),14 (link)]. Rat polyclonal antibodies against ARF6, ARNO and GST were created by immunization of rats with ARF6, ARNO and GST bacterial proteins, respectively.
Zhai Y., Chan W.W., Li W, & Lau K.F. (2022). ARNO is recruited by the neuronal adaptor FE65 to potentiate ARF6-mediated neurite outgrowth. Open Biology, 12(9), 220071.
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7

Antibody Identification for Protein Analysis

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Primary antibodies used for this study were as follows: rabbit anti-APOB (ab20737, Abcam); mouse anti-CHC (610500, BD Transduction Laboratories); rabbit anti-ERLIN1 (17311-1AP, proteintech); rabbit-anti ERLIN2 (14781-1-AP, proteintech); mouse anti-FLAG (F9291, Sigma); rabbit anti-FLAG (20543-1-AP, proteintech); mouse anti-MYC (sc-40, Santa Cruz Biotechnology); rabbit anti-MYC (06–549, Millipore); rabbit anti-TM6SF2 (AAS00444C, Antibody Verify). Horseradish peroxidase-conjugated goat anti-rabbit IgG (31460) and donkey anti-mouse IgG (715-035-150) antibodies were from Pierce and Jackson ImmunoResearch, respectively.
Li B.T., Sun M., Li Y.F., Wang J.Q., Zhou Z.M., Song B.L, & Luo J. (2020). Disruption of the ERLIN–TM6SF2–APOB complex destabilizes APOB and contributes to non-alcoholic fatty liver disease. PLoS Genetics, 16(8), e1008955.
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8

Immunoblot Analysis of Cellular Proteins

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Immunoblot was performed with standard protocol for these antibodies used at specified dilution: mouse anti-actin (Cell Signaling Technology #3700, 1:5000), mouse anti-HA (Cell Signaling Technology #2367, 1:1250), rabbit anti-FLAG (Proteintech #20543-1-AP, 1: 4000). PVDF membrane (Millipore) was blocked with fat-free skim milk in PBS-Tween 0.1% or TBS-Tween 0.1% followed by blotting with primary antibody overnight at 4 °C. Membrane was washed three times and then blotted with secondary antibody (HRP-conjugated mouse anti-rabbit (Sigma) or rabbit anti-mouse (Sigma), 1:10,000) for 1 h at room temperature. Signal was revealed with ECL Plus substrate (Thermo Fisher Scientific) after 3 washes.
Lambert-Lanteigne P., Young A, & Autexier C. (2023). Complex interaction network revealed by mutation of human telomerase ‘insertion in fingers’ and essential N-terminal domains and the telomere protein TPP1. The Journal of Biological Chemistry, 299(3), 102916.
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9

Biochemical Reagents for Cell Signaling

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Rabbit anti-TRPV1 (Alomone laboratory, catalog no. ACC-030), mouse anti-Kvβ1 (Santa Cruz, catalog no. sc-373986), rabbit anti-Na+/K+ ATPase (Abcam, catalog no. ab76020), mouse anti-GST (Abclonal, catalog no. AE001), mouse anti-β-actin (Transgen Biotech, catalog no. 6609-1), mouse anti-FLAG (Sigma, catalog no. F3165), mouse anti-FLAG (MBL, catalog no. M185-3L), rabbit anti-FLAG (Proteintech, catalog no. 20543-1-AP), mouse anti-HA (MBL, catalog no. M180-3), rabbit anti-HA (Earthox, catalog no. E02218002), mouse anti-GFP (Tianjin Sungene Biotech, catalog no. KM8009), rabbit anti-GFP (WB: Solarbio, catalog no. B1025F), anti-FLAG affinity gel (Bimake, catalog no. B23100), anti-HA affinity gel (Bimake, catalog no. B23301), ProteinIso GST resin (Transgen Biotech, catalog no. DP201-01), goat anti-mouse IgG (H + L) (Jackson Immunoresearch, catalog no. 115-035-003), and goat anti-rabbit IgG (H + L) (Jackson Immunoresearch, catalog no. 111-005-003) were purchased from the indicated manufactures. The EZ-Link Sulfo-NHS-LC-Biotin (catalog no. 21335) and high-capacity NeutrAvidin-agarose (catalog no. 29202) were ordered from Thermo Fisher Scientific.
Wang Y., Mo X., Ping C., Huang Q., Zhang H., Xie C., Zhong B., Li D, & Yao J. (2020). Site-specific contacts enable distinct modes of TRPV1 regulation by the potassium channel Kvβ1 subunit. The Journal of Biological Chemistry, 295(50), 17337-17348.
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