T6508

Manufactured by Merck Group

T6508 is a laboratory equipment product. It is a multi-channel pipette used for precise liquid handling in various laboratory applications. The device is designed to accurately measure and transfer small volumes of liquids.

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Lab products found in correlation

5 protocols using t6508

Antibody Peptide Mapping by LC-MS

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Reduced tryptic peptide
mapping was performed by first diluting 50 μg of antibody intermediate
to 10 mg/mL. Samples were denatured and reduced at 37 °C for
30 min in 20 μL of 6.3 M urea, 1 M guanadine HCl, 100 mM Tris
pH 8.0, and 5 mM dithiothreitol (DTT, Thermo Scientific) mixture.
Then, samples were alkylated with 15 mM iodoacetamide (IAM, Thermo
Scientific) for 30 min in the dark at room temperature. Subsequently,
the reaction was diluted with 3 volumes of 100 mM Tris pH 7.5 to allow
for trypsin digestion. Trypsin (V5280, Promega) was added at 1:12.5
protease: protein ratio and incubated at 37 °C for 3–4
h. The reaction was quenched by adding 5 μL of 10% TFA (T6508,
Sigma-Aldrich). The digests were analyzed by LC-MS using a Waters
ACQUITY UPLC system equipped with a Waters ACQUITY BEH C18 column
(1.7 mm, 2.1 × 150 mm), mobile phase A (0.02% TFA in HPLC water),
mobile phase B (0.02% TFA in acetonitrile) over a 90 min gradient,
and a Synapt G2 mass spectrometer (Waters).

Hines A.R., Edgeworth M., Devine P.W., Shepherd S., Chatterton N., Turner C., Lilley K.S., Chen X, & Bond N.J. (2023). Multi-Attribute Monitoring Method for Process Development of Engineered Antibody for Site-Specific Conjugation. Journal of the American Society for Mass Spectrometry, 34(7), 1330-1341.

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In Vivo FAM-Aβ Peptide Injection

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FAM-Aβ_(1–42) peptides (Anaspec, AS-23525) were dissolved in trifluoroacetic acid (Sigma, T6508), lyophilized in a SpeedVac, and resuspended in PBS/DMSO solution at 37 °C overnight to a final concentration of 2 μg/μl. FAM-Aβ aggregates (500 nl) were stereotaxically injected into hippocampi of control and MG^4E mice at P14 using the following coordinates (relative to the bregma): AP (anterior posterior), −2.25 mm; ML (medial lateral), ±1.80 mm; DV (dorsal ventral), −2.0 mm. Mice were perfused 18–19 h after FAM-Aβ injection.

Xu Z.X., Kim G.H., Tan J.W., Riso A.E., Sun Y., Xu E.Y., Liao G.Y., Xu H., Lee S.H., Do N.Y., Lee C.H., Clipperton-Allen A.E., Kwon S., Page D.T., Lee K.J, & Xu B. (2020). Elevated protein synthesis in microglia causes autism-like synaptic and behavioral aberrations. Nature Communications, 11, 1797.

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Inhibitor Library Preparation

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Chemicals were purchased from Selleckchem (Houston, TX, USA), Sigma-Aldrich (St. Louis, MO, USA), Cayman Chemical (Ann Arbor, MI, USA), and Med Chem Express (Monmouth Junction, NJ, USA). Included is the Selleckchem L-3500 screening library, the highly selective inhibitor library of 339 inhibitors covering 123 targets. Compounds were diluted in DMSO (Sigma-Aldrich, D2650) or water, depending on solubility, except for copanlisib which was diluted in 10% Trifluoroacetic acid in DMSO due (Sigma-Aldrich, T6508). Compounds were prepared as a 10 mM solution unless solubility constraints required lower concentrations.

Merrill N.M., Vandecan N.M., Day K.C., Palmbos P.L., Day M.L., Udager A.M., Merajver S.D, & Soellner M.B. (2020). MEK is a promising target in the basal subtype of bladder cancer. Oncotarget, 11(44), 3921-3932.

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Preparation and Use of Experimental Cell Culture Drugs

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Drugs for cell culture experiments were obtained as follows: buparlisib (SelleckChem; S2247), pictilisib (SelleckChem; S1065), copanlisib (SelleckChem; S2802), BX795 (SelleckChem; S1274), BX912 (SelleckChem; S1275), GSK2334470 (Tocris; 4143), bafilomycin A (Sigma-Aldrich; SML1661), 3-methyladenine (SelleckChem; S2767), autophinib (SelleckChem; S8596), and VPS34-IN1 (SelleckChem; 7980).
bafilomycin A was used at a concentration of 160 nM for all experiments. All other drug doses and incubation times are indicated in the figure legends or directly on the figures. Drugs were all dissolved in DMSO (Sigma-Aldrich; D2650), except for copanlisib and 3-methyladenine. copanlisib, which is not DMSO-soluble, was initially solubilized in a 10% trifluoroacetic acid (Sigma-Aldrich; T6508)/90% water solution to a concentration of 2 mM and then diluted to appropriate working stock concentrations. 3-methyladenine solutions were prepared fresh each time. 3-methyladenine powder was dissolved directly into cell culture medium to produce a 50 mM stock. 3-methyladenine solutions were heated to 55°C for 5 min, vortexed until clear, diluted to the appropriate concentration, and used immediately.
For all drug treatments, the cells were maintained in complete growth medium, with serum, according to the recipes specified above.

Endicott S.J., Ziemba Z.J., Beckmann L.J., Boynton DN J.r, & Miller R.A. (2020). Inhibition of class I PI3K enhances chaperone-mediated autophagy. The Journal of Cell Biology, 219(12), e202001031.

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Peptide Cleanup via SDB-RPS StageTips

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For all peptide samples, we used in-house made StageTips packed with SDB-RPS (2241, 3 M) material for desalting. About 1% trifluoroacetic acid (TFA; T6508; Sigma) was added into the reactions to stop digestion. For SDC-based digestion, centrifugation was performed before loading the peptides. For SCASP, no centrifugation was performed if there was no apparent pellet. The SDB-RPS StageTips were conditioned with 100 μl 100% acetonitrile (ACN) (3485; Sigma). The peptides were loaded into StageTips, followed by centrifugation at 4000g for 5 min. StageTips were washed twice with 100 μl 1% TFA/ isopropyl alcohol (I9030; Sigma), and then washed with 100 μl 0.2% TFA. Elution of peptides was performed using 80% ACN/5% ammonia water. All eluted materials were collected in glass vials (A3511040; CNW Technologies) and dried at 45 °C using a SpeedVac centrifuge (Eppendorf Concentrator Plus; 5305).

Gan G., Xu X., Chen X., Zhang X.F., Wang J, & Zhong C.Q. (2021). SCASP: A Simple and Robust SDS-Aided Sample Preparation Method for Proteomic Research. Molecular & Cellular Proteomics : MCP, 20, 100051.

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