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Flag antibody

Manufactured by Santa Cruz Biotechnology
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Sourced in United States

The Flag antibody is a widely used tool in molecular biology and biochemistry research. It is a monoclonal antibody that specifically recognizes the Flag epitope tag, which is a short amino acid sequence (DYKDDDDK) that can be fused to recombinant proteins. The Flag antibody allows for the detection and purification of Flag-tagged proteins, facilitating various experimental applications.

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Lab products found in correlation

Gfp antibody, by Santa Cruz Biotechnology (1 mentions) Pr 619, by Merck Group (1 mentions) Proteinase inhibitor cocktail, by Roche (1 mentions) G3bp1 antibody, by BD (1 mentions) Trisure reagent, by Meridian Bioscience (1 mentions) Protein g agarose beads, by Merck Group (1 mentions) Chip assay kit, by Merck Group (1 mentions)

Close spelling variants of this product

Anti-Flag antibody (23 citations) Anti-FLAG monoclonal antibody (5 citations) Rabbit anti‐flag antibody (3 citations) Monoclonal antibody against Flag-tag (2 citations)

6 protocols using flag antibody

1

Myocardin-Stat3 Interaction Assay

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Plasmid-based expression vectors encoding myc-tagged myocardin and flag-tagged Stat3 were cotransfected into cardiomyocytes. Lysates were collected 48 hours post-transfection. Then myc antibody (Santa Cruz) was used to precipitate myc-myocardin, and flag antibody (Santa Cruz) was used to precipitate flag-Stat3. The resulting mixture was washed, separated by SDS-PAGE, and transferred to an NC membrane (Pall). Samples were probed with myc antibody to visualize flag-epitope Stat3, stripped, and probed with flag antibody to visualize myc-epitope Myocardin.
Xiang Y., Liao X.H., Li J.P., Li H., Qin H., Yao A., Yu C.X., Hu P., Guo W., Gu C.J, & Zhang T.C. (2017). Myocardin and Stat3 act synergistically to inhibit cardiomyocyte apoptosis. Oncotarget, 8(59), 99612-99623.
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2

Butyrate Modulates HIF-1α Acetylation

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Butyrate (SIGMA, St Louis, CA, USA), sodium dichloroacetate (SIGMA), nicotinamide (SIGMA), rotenone (MERYER, Shanghai, China), TSA (CST, Danver, MA, USA), PDHA1α-phosphor-Ser293 Antibody (Novus Biologicals, Littleton, CO, USA), PDHA1α antibody (Abmart, Berkeley Heights, NJ, USA), Flag antibody (SIGMA), HA antibody (Santa Cruz, Santa Cruz, CA, USA) and the HIF-1α antibody (ABclone, Shanghai, China) were purchased. The pan-acetylation antibody was generated in-house.19 (link)
Xu S., Liu C.X., Xu W., Huang L., Zhao J.Y, & Zhao S.M. (2017). Butyrate induces apoptosis by activating PDC and inhibiting complex I through SIRT3 inactivation. Signal Transduction and Targeted Therapy, 2, 16035-.
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3

Immunoprecipitation of GFP Fusion Proteins

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Cells were washed twice with PBS and lysed with IP lysis buffer (25 mM Tris-HCl, pH 7.4, 150 mM NaCl, 1% NP-40, 1 mM EDTA, 5% glycerol; Thermo Scientific 87787) supplemented with 20 mM N-ethylmaleimide (NEM) (Sigma-Aldrich E3876), 50 μM PR-619 (Sigma-Aldrich, 662141) and proteinase inhibitor cocktail (Roche 1183617001). Lysates were centrifuged at 4°C for 15 min at 20,000g. The supernatants were incubated with normal mouse IgG (Santa Cruz Biotechnology sc-2025), GFP antibody (Santa Cruz Biotechnology sc-9996), VCP antibody (Santa Cruz Biotechnology sc-57492), FLAG antibody (Santa Cruz Biotechnology sc-166355), or G3BP1 antibody (BD 611127) conjugated with protein A/G magnetic beads (Thermo Fisher Scientific 88803) at 4°C overnight. Beads were washed three times with wash buffer (50 mM Tris-HCl, pH 7.5, 500 mM NaCl, 0.05% Tween-20, 20 mM NEM and 50 μM PR-619) and treated with 0.1 M glycine pH 2.7 (Teknova G4527) at room temperature for 10 min to elute proteins. The resulting samples were analyzed by immunoblotting.
Gwon Y., Maxwell B.A., Kolaitis R.M., Zhang P., Kim H.J, & Taylor J.P. (2021). Ubiquitination of G3BP1 mediates stress granule disassembly in a context-specific manner. Science (New York, N.Y.), 372(6549), eabf6548.
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4

Flag-Tag Protein Immunoprecipitation

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Transfected cells were lysed in cell lysis buffer (50 mM Tris-HCl pH 8.0, 150 mM NaCl, 1 mM EDTA, 1% NP40, 10% glycerol with protease inhibitor cocktail) for 1 h. Whole-cell extracts were collected and precleared. Beads coated with FLAG antibody (CST #2368), OCT4 antibody (sc-8628, Santa Cruz Biotechnology), and ZNF207 antibody (PA5-30641, ThermoFisher Scientific) were incubated with precleared whole-cell extracts at 4 °C overnight. The beads were washed with cell lysis buffer four times. Finally, the beads were boiled in 2x sample buffer for 10 min. The eluents were analyzed by Western blot.
Fang F., Xia N., Angulo B., Carey J., Cady Z., Durruthy-Durruthy J., Bennett T., Sebastiano V, & Reijo Pera R.A. (2018). A distinct isoform of ZNF207 controls self-renewal and pluripotency of human embryonic stem cells. Nature Communications, 9, 4384.
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5

Identification of miRNA-protein complexes

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A Flag-Ago2 expression construct was combined with mimic miRNAs of miR-105 or miR93-3p (TOOLS), transfected into 293 T cells and incubated for 48 hours. The cell lysates were harvested using RIPA buffer, containing protease inhibitors and RNase inhibitor. Flag antibody (Santa Cruz Technology, sc-9996) was pre-incubated with Protein G-agarose beads (EMD Millipore, 16-266) overnight at 4 °C and washed three times with IP wash buffer. The cell lysates were incubated with pre-conjugated antibodies overnight at 4 °C and washed three times with IP wash buffer. The pellet from immunoprecipitation was resolved in TRIsure reagent (BIOLINE, Taunton, MA, USA, BIO-38033) for RNA extraction and PCR.
Li H.Y., Liang J.L., Kuo Y.L., Lee H.H., Calkins M.J., Chang H.T., Lin F.C., Chen Y.C., Hsu T.I., Hsiao M., Ger L.P, & Lu P.J. (2017). miR-105/93-3p promotes chemoresistance and circulating miR-105/93-3p acts as a diagnostic biomarker for triple negative breast cancer. Breast Cancer Research : BCR, 19, 133.
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6

ChIP Assay for Protein-DNA Interactions

Cited 1 time
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A ChIP assay was performed using a ChIP Assay Kit (Millipore) according to the manufacturer’s instructions. Briefly, LN-229 cells were cross-linked and lysed, and DNA was sheared into 200–800 bp fragments using sonication. Precleared chromatin was immunoprecipitated with a Flag antibody (Santa Cruz), and DNA was isolated after reverse cross-linking for quantitative real-time PCR (qRT-PCR). The relevant primer sequences are presented in Table S3 in the Supplemental Materials. ChIP analyses were then performed as previously described33 (link).
Deng Q., Hou J., Feng L., Lv A., Ke X., Liang H., Wang F., Zhang K., Chen K, & Cui H. (2018). PHF19 promotes the proliferation, migration, and chemosensitivity of glioblastoma to doxorubicin through modulation of the SIAH1/β–catenin axis. Cell Death & Disease, 9(11), 1049.
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