BALB/c (IMSR catalog no. JAX:000651, RRID:IMSR_JAX:000651) mice were sublethally X-ray irradiated with 300 cGy (CIXD irradiator, Xstrahl) and injected with 5 × 105 Ba/F3-FLT3/ITD-Luc cells via tail vein. Intraperitoneal injections were performed using PBS vehicle with human AGP (Sigma) at 20 mg/mL. Midostaurin was suspended in 30% (w/v) Cremophor EL, 30% (w/v) PEG 400, 10% ethanol, and 10% glucose (all Sigma) and instilled once daily by oral gavage (10 (link), 11 (link)). These drug doses have previously been demonstrated to be effective in mice. Mifepristone (Sigma) was dissolved in 50% ethanol and 50% (2-hydroxypropyl)-β-cyclodextrin (Sigma) and gavaged every 48 hours. Mice were imaged by intraperitoneal injection of luciferin (3 mg) and visualized on an IVIS Spectrum imager (Caliper LifeSciences) using Living Image software for analysis on day 5 (to monitor engraftment) and on day 9 (to assess drug effect). Animal husbandry and procedures were conducted in accordance with the policy of the Johns Hopkins University School of Medicine Animal Care and Use Committee.
Human agp
Manufactured by Merck Group
Sourced in United States
Human AGP is a laboratory equipment product produced by Merck Group. It is designed to perform specific functions related to laboratory analysis and experimentation. The core function of this product is to assist in the research and development processes conducted in a scientific setting. No further details or interpretations about the intended use of this product are provided.
Automatically generated - may contain errors
Lab products found in correlation
Mifepristone, by Merck Group (2 mentions)
Cixd irradiator, by Xstrahl (2 mentions)
Cremophor el, by Merck Group (2 mentions)
2 hydroxypropyl β cyclodextrin, by Merck Group (2 mentions)
Peg400, by Merck Group (2 mentions)
Human hb, by Merck Group (1 mentions)
Q exactive plus orbitrap mass spectrometer, by Thermo Fisher Scientific (1 mentions)
Q exactive mass spectrometer, by Thermo Fisher Scientific (1 mentions)
Potassium hexafluorophosphate, by Merck Group (1 mentions)
Sodium chloride (nacl), by Fujifilm (1 mentions)
V 660, by Jasco (1 mentions)
L ascorbic acid sodium salt, by Fujifilm (1 mentions)
Tris hcl buffer, by Fujifilm (1 mentions)
4 protocols using human agp
1
Xenograft Model of Acute Myeloid Leukemia
2
Evaluating FLT3/ITD Leukemia Treatment
3
Native MS Analysis of Glycoproteins
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Extended experimental and method details can be found in SI Appendix . Human AGP (product number G9885), human Hp phenotype 1-1 (product number H0138), and human Hb (product number H7379) were purchased from Sigma-Aldrich. For native MS analysis, glycoproteins were dissolved in water, buffer exchanged to 200 mM ammonium acetate (pH 7.6), and analyzed on a modified Q-Exactive mass spectrometer (Thermo Fisher Scientific) (56 (link)) and a modified Q-Exactive Plus Orbitrap mass spectrometer (57 (link)). The raw native MS spectra were deconvoluted using UniDec software to produce zero-charge spectra. Protein and glycan mass calculations were based on amino acid and monosaccharide average residue masses. AGP–warfarin and Hp–Hb binding experiments were performed in ammonium acetate buffer (pH 7.6). Protein structures of AGP [Protein Data Bank (PDB) ID code 3KQ0] (17 (link)) and Hp–Hb complex (PDB ID code 4WJG) (43 (link)) were retrieved from PDB and processed using University of California, San Francisco Chimera program (version 1.22) (58 (link)).
4
Enantioselective Enzymatic Biosynthesis
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The enantiomeric excess (S)-CypL (the enantiomeric mixture of CypL; lot number: L1816) was from NanoLight Technology, Prolume (Pinetop, AZ, USA) (see Supplementary Figure S1 and Table S1 ). Human AGP (alpha 1-acid glycoprotein form human plasma) and potassium hexafluorophosphate were from Sigma-Aldrich (St. Louis, MO, USA). Tris–HCl buffer, l -ascorbic acid sodium salt, and sodium chloride were from FUJIFILM Wako Pure Chemical Corporation (Osaka, Japan). All materials were used without further purification. Racemic CypL and (S)-CypL were prepared according to the method reported previously [19 (link),20 (link)]. A recombinant CypLase from C. noctiluca was prepared using plant cells according to the method reported previously (see Supplementary Figure S2 ) [14 (link)]. The concentrations of CypL solutions were determined spectrophotometrically using a spectrophotometer (V-660; Jasco, Tokyo, Japan), according to the reported molar absorption coefficient [24 (link)]. The concentrations of Human AGP and recombinant CypLase solutions were determined by SDS-PAGE analysis or by using the corresponding molar extinction coefficient at 280 nm, as calculated via the peptide property calculator available at https://www.biosyn.com/peptidepropertycalculator/peptidepropertycalculator.aspx (accessed on 6 October 2023).
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