Rabbit α flag

Manufactured by Merck Group

Rabbit α-FLAG is a laboratory reagent used for the detection and purification of proteins tagged with the FLAG peptide sequence. It is a polyclonal antibody raised in rabbits against the FLAG tag, which is a short amino acid sequence commonly used as a molecular tag in recombinant proteins. This antibody can be used in various immunological techniques, such as immunoprecipitation, Western blotting, and immunoaffinity chromatography, to identify and isolate FLAG-tagged proteins.

Automatically generated - may contain errors

Lab products found in correlation

Close spelling variants of this product

α-FLAG (120 citations)

6 protocols using rabbit α flag

Comprehensive Antibody Reagents for Western Blotting

Check if the same lab product or an alternative is used in the 5 most similar protocols

Mouse α-GST (sc-138; 1:1,000), goat α-SRSF1 (sc-10254; 1:1,000), rabbit α-U2AF65 (sc-48804; 1:5,000), goat α-actin (sc-1616; 1:1,000), mouse α-tubulin (sc-8035; 1:3,000), goat α-BLM (sc-7790; 1:1,000), mouse α-SUMO1 (sc-5308; 1:1,000) and rabbit α-RNAPII (sc-899; 1:1,000) were purchased from Santa Cruz. Mouse α-RNAPII phospho-CTD (phospho S2; H5; ab24758; 1:5,000) and rabbit α-THOC5 (ab86070; 1:5,000) were purchased from Abcam. Rabbit α-Histone H4 (no. 2592; 1:1,000) and rabbit α-PIAS1 (no. 3550; 1:1,000) were purchased from Cell Signaling. Mouse α-NWSHPQFEK tag (StrepII tag; A01732; 1:3,000) was from GeneScript. Mouse α-p84 (GTX70220; 1:5,000) was from GeneTex and rabbit α-TOP1 (no. 3552–1; 1:5,000) was from Epitomics. Rabbit α-FLAG (F7425; 1:3,000) was purchased from Sigma-Aldrich. Mouse α-TOP1 (1:5,000) was kindly provided by Dr Yung-Chi Cheng (Yale School of Medicine). Mouse α-PCNA (1:5,000) was kindly provided by Dr Stephen West (Cancer Research UK). Rabbit α-RECQ5 (1:3,000) was generated as described^{15 (link)}. Mouse α-DNA-RNA hybrid [S9.6] was kindly provided by Dr Stephen H. Leppla National Institutes of Health (NIH).

Li M., Pokharel S., Wang J.T., Xu X, & Liu Y. (2015). RECQ5-dependent SUMOylation of DNA topoisomerase I prevents transcription-associated genome instability. Nature communications, 6, 6720.

+ Open protocol

+ Expand

Immunofluorescence Staining of HEp-2 Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

HEp-2 cells were seeded in 24-well format on glass cover slides. After transfecting with jetPrime (Polyplus-transfection), cells were fixed with 3% paraformaldehyde (PFA) in PBS for 10 min at room temperature and permeabilized with 0.1% or 0.5% Triton X-100 in PBS for 10 min at room temperature. Staining was performed in PBS with 5% goat serum for 1 h at RT with mouse anti-MxA (ɑ-MxA) (hybridoma ab143, 1:5, in house), mouse α-MxB (sc-271527, 1:50; Santa Cruz Biotechnology), mouse α-FLAG M2 (F1804, 1:1,000; Sigma), rabbit α-FLAG (F7425, 1:1,000; Sigma), or mouse α-NP (hybridoma HB65, 1:5, ATCC).

Steiner F, & Pavlovic J. (2020). Subcellular Localization of MxB Determines Its Antiviral Potential against Influenza A Virus. Journal of Virology, 94(22), e00125-20.

+ Open protocol

+ Expand

Immunoblotting with Epitope-Tagged Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols

The antibodies used were mouse α-Myc (Santa Cruz Biotechnology), mouse monoclonal α-FLAG M2 (Sigma-Aldrich), rabbit α-FLAG (Sigma-Aldrich), and α-V5-HRP conjugate (Invitrogen).

Halabi S., Sekine E., Verstak B., Gay N.J, & Moncrieffe M.C. (2016). Structure of the Toll/Interleukin-1 Receptor (TIR) Domain of the B-cell Adaptor That Links Phosphoinositide Metabolism with the Negative Regulation of the Toll-like Receptor (TLR) Signalosome. The Journal of Biological Chemistry, 292(2), 652-660.

+ Open protocol

+ Expand

Western Blot Analysis of ER Quality Control

Check if the same lab product or an alternative is used in the 5 most similar protocols

Primary antibodies used were as follows: rabbit α-TXNDC11 (Abcam, ab188329; 1:5,000), rabbit α-FLAG (Sigma-Aldrich, F7425; 1:10,000), mouse M1 α-FLAG (Sigma-Aldrich, F3040; 1:10,000), rabbit α-EDEM2 (Sigma-Aldrich, E1728; 1:5,000), rabbit α-EDEM3 (Sigma-Aldrich, E0409; 1:5,000), rabbit α-PDI (Cell Signaling, #2446; 1:5,000), rabbit α-GANAB (GeneTex, GTX102237; 1:5,000), rabbit α-SEL1L (Santa Cruz Biotechnology, sc-48080; 1:2,000), mouse α-calnexin (AF8, a kind gift from M. Brenner, Harvard Medical School; 1:10,000) and mouse α-β-actin (Sigma-Aldrich, A5316; 1:10,000).

Timms R.T., Menzies S.A., Tchasovnikarova I.A., Christensen L.C., Williamson J.C., Antrobus R., Dougan G., Ellgaard L, & Lehner P.J. (2016). Genetic dissection of mammalian ERAD through comparative haploid and CRISPR forward genetic screens. Nature Communications, 7, 11786.

+ Open protocol

+ Expand

Comprehensive Molecular Assay Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

Hydroxyurea (Catalog No. H8627), doxorubicin hydrochloride (Catalog No. D1515), thioridazine hydrochloride (Catalog No. T9025), KU-55933 (Catalog No SML1109) and G418 disulfate salt (Catalog No. A1720) were purchased from Sigma. Dulbecco’s modified Eagle’s medium (DMEM) was obtained from Life Technologies (Catalog No. 12100-046), fetal bovine serum (FBS) was obtained from Atlanta Biologicals (Catalog No. 900108). Enhanced chemiluminescence solution was obtained from Thermo-Scientific (Catalog number No. 32106).
Antibodies used in this study were: rabbit α-Actin (Ab1801, Abcam), rabbit α-Rad9 phospho-328 (AP3225a, Abgent), rabbit α-Chk1 phospho-317 (AP3070a, Abgent), rabbit α-Chk1 phospho-345 (sc-17922, Santa Cruz Biotechnology), goat α-Rad9 (sc-10465, Santa Cruz Biotechnology), mouse α-Rad9 (sc-8324, Santa Cruz Biotechnology), donkey α-goat IgG-HRP (sc-2020, Santa Cruz Biotechnology), rabbit α-TLK1 phospho-695 (4121S, Cell Signaling), α-rabbit IgG-HRP (7074S, Cell Signaling), α-mouse IgG-HRP (7076, Cell Signaling), rabbit α-TLK1 (GTX102891, GeneTex), mouse α- H2A.X phospho-139 (05-636, Millipore), mouse α-Flag (F1804, Sigma) and Rabbit α-Flag (F7425, Sigma).

Awate S, & De Benedetti A. (2016). TLK1B mediated phosphorylation of Rad9 regulates its nuclear/cytoplasmic localization and cell cycle checkpoint. BMC Molecular Biology, 17, 3.

+ Open protocol

+ Expand

Western Blot Protein Detection Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

Proteins were separated by polyacrylamide gel electrophoresis and transferred to nitrocellulose membranes (BioRad). Membranes were dried, rehydrated with Tris-buffered saline (TBS) and blocked with TBS blocking buffer (927-60001; Li-Cor) before incubation with primary antibody diluted in TBS blocking buffer overnight. Membranes were washed and incubated with secondary antibodies diluted in the same buffer for 1 hour. The antibodies used in this study are: mouse α-FLAG (M2; Sigma) 1:2 500; rabbit α-FLAG (SAB4301135; Sigma) 1:2 500; rat α-HA (clone 3F10; Roche Diagnostics); mouse α-Myc (9E10; Santa Cruz Biotechnology) 1:2 500; goat α-mouse (926-68020; Licor) 1:5 000; goat α-rabbit (925-32211; Licor) 1:5 000; goat α-rat (926-32219; Licor) 1:5 000. Blots were imaged with a Li-Cor Odyssey and Image Studio software.

Delyea C., Luo S.Y., Dubrule B.E., Julien O., & Bhavsar A.P. (2021). NOD1 is super-activated through spatially-selective ubiquitination by the Salmonella effector SspH2.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!