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Anti flag

Manufactured by Applied Biological Materials
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Sourced in United States, China

Anti-Flag is a laboratory product designed to detect and purify proteins with a specific epitope tag. It functions as an affinity reagent, enabling the identification and isolation of target proteins in various research applications.

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Lab products found in correlation

Anti flag hrp, by Merck Group (3 mentions) Anti ha, by Applied Biological Materials (2 mentions) Protein a g agarose, by Santa Cruz Biotechnology (2 mentions) Anti v5, by Thermo Fisher Scientific (2 mentions) Anti ephrinb2, by Abcam (2 mentions) Anti adam10, by Abcam (2 mentions) Anti flotillin 1, by Santa Cruz Biotechnology (2 mentions) Anti ha hrp, by Roche (2 mentions) Anti ephrinb2, by R&D Systems (2 mentions) Anti erk2, by Santa Cruz Biotechnology (2 mentions) Anti ubiquitin, by Merck Group (1 mentions) Anti acetylated lysine, by Cell Signaling Technology (1 mentions) Anti α tubulin, by Merck Group (1 mentions) Anti myc, by Applied Biological Materials (1 mentions) Anti p300, by BD (1 mentions) Anti ki67, by Abcam (1 mentions) Anti ha, by Merck Group (1 mentions) Anti flag, by Merck Group (1 mentions) Anti ha hrp, by Merck Group (1 mentions) Anti gst, by Santa Cruz Biotechnology (1 mentions)

5 protocols using anti flag

1

Immunoblot and Immunostaining Antibodies

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Antibodies used for the immunoblot and immunostaining were anti-p300 (BD Biosciences, San Jose, CA, USA), anti-α-tubulin (Sigma), anti-myc, anti-Flag, anti-GFP, anti-HA (abm, Richmond, Canada), anti-ubiquitin (Millipore, Billerica, MA, USA), anti-acetylated lysine (Cell signaling, Danvers, MA, USA). Anti-HIPK2 antibody was a kind gift of Dr. K. Isono (RIKEN, Japan) and described previously27 (link).
Choi J.R., Lee S.Y., Shin K.S., Choi C.Y, & Kang S.J. (2017). p300-mediated acetylation increased the protein stability of HIPK2 and enhanced its tumor suppressor function. Scientific Reports, 7, 16136.
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2

Evaluating ZNF471 Expression in ESCC

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To evaluate the expression levels of ZNF471 in ESCC tissues, tissue microarrays were constructed through paraffin embedding. IHC was performed according to a previously described procedure 58 (link), 59 (link). Anti-Flag was obtained from Abm (#G188; Abm, Richmond, BC), anti-MAPK10 was obtained from Bioss (bs-2997R, Bioss, Beijing, China), and anti-Ki-67 was obtained from Abcam (ab15580; Abcam, Cambridge, UK). Sections were incubated with primary antibody (1:50 dilution) overnight at 4°C and then with secondary antibody (1:2000 dilution) at 37°C for 30 min. Finally, the slides were counterstained with hematoxylin.
Sun R., Xiang T., Tang J., Peng W., Luo J., Li L., Qiu Z., Tan Y., Ye L., Zhang M., Ren G, & Tao Q. (2020). 19q13 KRAB zinc-finger protein ZNF471 activates MAPK10/JNK3 signaling but is frequently silenced by promoter CpG methylation in esophageal cancer. Theranostics, 10(5), 2243-2259.
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3

Antibody Selection for Protein Detection

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The following antibodies were used: anti-FLAG, anti-FLAG-HRP, anti-HA, anti-HA-HRP (Sigma Aldrich), anti-FLAG, anti-HA, anti-GST (abm), anti-GST, anti-His (Santa cruz biotechnology), anti-H1, H2A, H2B (Abcam) and anti-H3, H4, H3K9Ac, H3K27Ac, H3K4me3, H3K9me2, H3K27me2 (Upstate).
Lee W.K, & Cho M.H. (2016). Telomere-binding protein regulates the chromosome ends through the interaction with histone deacetylases in Arabidopsis thaliana. Nucleic Acids Research, 44(10), 4610-4624.
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4

Immunoprecipitation and Western Blot Analysis

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Immunoprecipitation and Western blot analysis were performed as described previously with a minor modification55 (link). Briefly, lysates of HT29 cells, oocytes, or embryos were prepared with cold lysis buffer. Immunoprecipitations were conducted on HT29 cell lysates or 15 oocyte/embryo equivalents at 4°C for 4–16 hrs with the anti-HA, anti-Flag (Applied Biological Materials), anti-ephrinB2 (R&D systems AF496) or anti-His antibodies, and protein-A/G agarose (Santa Cruz Biotechnology). Immunoprecipitates were washed with either RIPA or lysis buffer three times. Immunoblotting was conducted using anti-Flag-HRP (Sigma), anti-HA-HRP (Roche), anti-ephrinB2 (Abcam 75868), anti-Flotillin1 (Santa Cruz Biotechnology SC74566), anti-ADAM10 (Abcam, ab1997) anti-Erk2 (Santa Cruz Biotechnology), or anti-V5 (Invitrogen) antibodies in 5 % milk. anti-Flag-HRP and anti-HA-HRP were diluted 1:2,000, anti-ephrinB2 and anti-Flotillin-1 were diluted 1:1,000, anti-Erk2 was diluted 1:10,000, and anti-V5 was diluted 1:5,000.
Ji Y.J., Hwang Y.S., Mood K., Cho H.J., Lee H.S., Winterbottom E., Cousin H, & Daar I.O. (2014). EphrinB2 Affects Apical Constriction in Xenopus Embryos and is Regulated by ADAM10 and Flotillin-1. Nature communications, 5, 3516.
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5

Immunoprecipitation and Western Blot Analysis

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Immunoprecipitation and Western blot analysis were performed as described previously with a minor modification55 (link). Briefly, lysates of HT29 cells, oocytes, or embryos were prepared with cold lysis buffer. Immunoprecipitations were conducted on HT29 cell lysates or 15 oocyte/embryo equivalents at 4°C for 4–16 hrs with the anti-HA, anti-Flag (Applied Biological Materials), anti-ephrinB2 (R&D systems AF496) or anti-His antibodies, and protein-A/G agarose (Santa Cruz Biotechnology). Immunoprecipitates were washed with either RIPA or lysis buffer three times. Immunoblotting was conducted using anti-Flag-HRP (Sigma), anti-HA-HRP (Roche), anti-ephrinB2 (Abcam 75868), anti-Flotillin1 (Santa Cruz Biotechnology SC74566), anti-ADAM10 (Abcam, ab1997) anti-Erk2 (Santa Cruz Biotechnology), or anti-V5 (Invitrogen) antibodies in 5 % milk. anti-Flag-HRP and anti-HA-HRP were diluted 1:2,000, anti-ephrinB2 and anti-Flotillin-1 were diluted 1:1,000, anti-Erk2 was diluted 1:10,000, and anti-V5 was diluted 1:5,000.
Ji Y.J., Hwang Y.S., Mood K., Cho H.J., Lee H.S., Winterbottom E., Cousin H, & Daar I.O. (2014). EphrinB2 Affects Apical Constriction in Xenopus Embryos and is Regulated by ADAM10 and Flotillin-1. Nature communications, 5, 3516.
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