Rabbit anti flag ab

Manufactured by Cell Signaling Technology

Rabbit anti-FLAG-AB is a primary antibody that recognizes the FLAG epitope tag. It is commonly used to detect and purify recombinant proteins tagged with the FLAG sequence.

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Lab products found in correlation

Ab21623, by Abcam (1 mentions) Pgex 4t 1, by GE Healthcare (1 mentions) Ab6721, by Abcam (1 mentions) Ab18184, by Abcam (1 mentions) Taq dna ligase, by New England Biolabs (1 mentions) T5 exonuclease, by New England Biolabs (1 mentions) Phusion dna polymerase, by New England Biolabs (1 mentions) Restriction enzyme, by New England Biolabs (1 mentions) Ab6728, by Abcam (1 mentions) α actinin, by Santa Cruz Biotechnology (1 mentions) C maf, by Santa Cruz Biotechnology (1 mentions) Halt protease and phosphatase inhibitor cocktail, by Thermo Fisher Scientific (1 mentions) Gapdh, by Merck Group (1 mentions)

2 protocols using rabbit anti flag ab

Bacterial Expression Vectors for Protein Modification

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For expression in bacterial cells pRSFDuet-1 (Merck/Sigma-Aldrich, Novagen) and a vector derived from pGEX-4T-1 (GE Healthcare) were used and modified as described above (Supplementary Data 4). Mutations were introduced by site-directed mutagenesis according to QuickChange protocol (Supplementary Table 8). For cloning, Phusion-DNA-polymerase, Taq-DNA-ligase, T5 exonuclease, and restriction enzymes were used (New England Biolabs). Anti-AcK-, anti-His₆-, anti-FLAG-primary antibodies and suitable HRP-coupled secondary antibodies were purchased from Abcam, Cell Signaling Technologies and Invitrogen (rabbit anti-AcK-AB: abcam, ab21623/dilution: 1:1000 in 3-5% (w/v) milk; mouse anti-His₆-AB: abcam, ab18184/dilution: 1:000 in 3-5% (w/v) milk; rabbit anti-FLAG-AB: Cell Signaling Technology, CST14793/dilution: 1:1000 in 3–5% (w/v) milk; mouse anti-FLAG-AB: Invitrogen, FG4R/dilution: 1:2000 in 3% (w/v) milk; goat anti-rabbit-HRP-AB: abcam, ab6721/dilution: 1:10000 in 3-5% milk; rabbit anti-mouse-HRP-AB: abcam, ab6728/dilution: 1:10000 in 1-5% (w/v) milk).

Kremer M., Schulze S., Eisenbruch N., Nagel F., Vogt R., Berndt L., Dörre B., Palm G.J., Hoppen J., Girbardt B., Albrecht D., Sievers S., Delcea M., Baumann U., Schnetz K, & Lammers M. (2024). Bacteria employ lysine acetylation of transcriptional regulators to adapt gene expression to cellular metabolism. Nature Communications, 15, 1674.

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Western Blot Analysis of Protein Targets

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Abs for KLF13, c-Maf, and α-actinin were purchased from Santa Cruz Biotechnology (Santa Cruz, CA), and GAPDH was purchased from Millipore (Billerica, MA). Mouse anti-V5 Ab was purchased from Invitrogen (Carlsbad, CA). Rabbit anti-FLAG Ab was purchased from Cell Signaling (Danvers, MA). For Western blot analysis, cells were lysed in cell lysis buffer (150 mM NaCl; 1 mM EDTA; 1 mM ethylene glycol-bis(β-aminoethyl ester)-N,N,N′,N′-tetraacetic acid; 1% Triton; 2.5 mM sodium pyrophosphate; 1 mM β-glycerolphosphate; 1 mM Na₂VO₄, in 20 mM Tris, pH 7.4) with Halt Protease and Phosphatase Inhibitor Cocktail (Thermo Scientific, Rockford, IL) and incubated on ice for 30 min. After centrifugation, the supernatant proteins (containing 25–50 μg protein) were separated by SDS-PAGE and transferred onto polyvinylidene difluoride membranes. Appropriate primary Abs and secondary Abs were used for Western blots.

Kwon S.J., Crespo-Barreto J., Zhang W., Wang T., Kim D.S., Krensky A, & Clayberger C. (2014). KLF13 Cooperates with c-Maf To Regulate IL-4 Expression in CD4+ T Cells. Journal of immunology (Baltimore, Md. : 1950), 192(12), 5703-5709.

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