Anti flt3 s18 antibody

Manufactured by Santa Cruz Biotechnology

The Anti-FLT3 S18 antibody is a research-use-only product developed by Santa Cruz Biotechnology. It is a mouse monoclonal antibody raised against a synthetic peptide corresponding to amino acids 18-32 of the FLT3 protein. The antibody can be used to detect the FLT3 protein by various immunological methods.

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Anti-FLT3 antibody Anti-FLT3

2 protocols using anti flt3 s18 antibody

Investigating Mutant Kinase Signaling

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Exponentially growing Molm14, HB119, or Ba/F3 cells stably expressing mutant isoforms were plated in RPMI medium 1640 + 10% (vol/vol) FCS supplemented with PKC412 or Star 27 at the indicated concentration. HMC1.2 cells were cultured and treated in IMDM + 10% FCS. After a 90-min incubation, the cells were washed in PBS, lysed, and processed as previously described. Immunoblotting was performed using anti-phospho-FLT3, anti-phospho-KIT, anti-phospho-STAT5, anti-STAT5, anti-phospho-ERK, anti-ERK, anti-phospho-S6, anti-S6, anti-KIT (Cell Signaling, Beverley, MA) and anti-FLT3 S18 antibody (Santa Cruz Biotechnology, Santa Cruz, CA).

Warkentin A.A., Lopez M.S., Lasater E.A., Lin K., He B.L., Leung A.Y., Smith C.C., Shah N.P, & Shokat K.M. (2014). Overcoming myelosuppression due to synthetic lethal toxicity for FLT3-targeted acute myeloid leukemia therapy. eLife, 3, e03445.

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FLT3 Mutant Isoform Signaling Analysis

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Exponentially growing Ba/F3 cells stably expressing FLT3 mutant isoforms were plated in RPMI medium 1640 + 10% FCS supplemented with kinase inhibitor at the indicated concentration. After a 90-minute incubation, the cells were washed in phosphate buffered saline (PBS) and lysed in buffer (50mM HEPES, pH 7.4, 10% glycerol, 150mM NaCl, 1% Triton X-100, 1mM EDTA, 1mM EGTA, 1.5mM MgCl₂) supplemented with protease and phosphatase inhibitors. The lysate was clarified by centrifugation and quantitated by BCA assay (Thermo Scientific; Rockford, IL). Protein was subjected to sodium dodecyl sulfate polyacrylamide electrophoresis and transferred to nitrocellulose membranes. Immunoblotting was performed using anti-phospho-FLT3 (3464), anti-phospho-STAT5 (9351), anti-STAT5 (9363), anti-phospho-S6 (2211), anti-S6 (2317), anti-phospho-ERK (9101), anti-ERK (9102) (Cell Signaling; Beverly, MA), and anti-FLT3 S18 antibody (Santa Cruz Biotechnology; Santa Cruz, CA). Data shown in figures is representative of 3 technical replicates.

Smith C.C., Zhang C., Lin K., Lasater E.A., Zhang Y., Massi E., Damon L.E., Pendleton M., Bashir A., Sebra R., Perl A., Kasarskis A., Shellooe R., Tsang G., Carias H., Powell B., Burton E.A., Matusow B., Zhang J., Spevak W., Ibrahim P.N., Le M.H., Hsu H.H., Habets G., West B.L., Bollag G, & Shah N.P. (2015). Characterizing and Overriding the Structural Mechanism of the Quizartinib-resistant FLT3 “Gatekeeper” F691L Mutation with PLX3397. Cancer discovery, 5(6), 668-679.

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