One mg of each of the three purified proteins; serum albumin (BSA, Sigma), myoglobin from horse (MYG, Sigma), and alcohol dehydrogenase from yeast (ADH1, Sigma) were dissolved in separate 1 mL aliquots of 170 mM Na2HPO4 pH 8. Chemical cross-linking with iqPIR was carried out by adding either the SH or RH iqPIR from a concentrated stock in DMSO to the protein containing solution at a final concentration of 1 mM and allowing the reaction to proceed for 30 min at 22 °C with constant mixing at 600 rpm on a Thermomixer. Aliquots of the cross-linked proteins were then mixed at the following SH/RH ratios to generate three different mixed samples each containing 100 μg total of each of the three proteins; mix 1 = BSA 4:1, Myg 1:10, ADH 1:2, mix 2 = BSA 1:10, Myg 2:1, ADH 4:1, mix 3 = BSA 2:1, Myg 1:4, and ADH 10:1. Disulfides in the three mixed samples were reduced by adding TCEP to 5 mM from a 500 mM stock and mixing at room temperature for 30 min. Reduced thiols were alkylated with 10 mM iodoacetamide added from a 1 M stock. Proteins were digested with a 1:200 ratio of trypsin to protein at 37 °C for 16 h. The resulting peptide samples were desalted by solid phase extraction using Waters C18 SepPak cartridges.
Adh 1
Manufactured by Merck Group
Sourced in United States
ADH-1 is a laboratory equipment product manufactured by Merck Group. It is a device used for the detection and analysis of alcohol dehydrogenase (ADH) enzyme activity in various samples. The core function of ADH-1 is to measure the catalytic activity of the ADH enzyme, which is essential for the oxidation of alcohols to aldehydes or ketones. The detailed technical specifications and intended applications of this product are not available for inclusion in an unbiased, factual description.
Automatically generated - may contain errors
Lab products found in correlation
Sep pak c18 cartridge, by Waters Corporation (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
Kg 501, by Merck Group (1 mentions)
Rucaparib, by Selleck Chemicals (1 mentions)
Prexasertib, by MedChemExpress (1 mentions)
Olaparib, by Merck Group (1 mentions)
Prexasertib, by Merck Group (1 mentions)
Rucaparib, by Merck Group (1 mentions)
Adh 1, by MedChemExpress (1 mentions)
Olaparib, by Selleck Chemicals (1 mentions)
Hydroxyurea hu, by Merck Group (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Cx546, by Merck Group (1 mentions)
4 protocols using adh 1
1
Cross-linking and Trypsin Digestion of Proteins
2
Intrathecal Injection of CREB and N-cadherin Inhibitors
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The CREB inhibitor KG-501 (Cat. # 70485, Sigma, USA) and N-cadherin inhibitor ADH-1 (Cat. # HY-13541, MedChemExpress, USA) were dissolved in dimethyl sulfoxide (DMSO) and diluted with 0.9% saline. Intrathecal injection of the drugs in a volume of 6 µL was performed using a 10 µL Hamilton microsyringe with a 27-gauge needle on day 20 after plantar incision as described previously.22 (link),23 (link) The accurate placement of the needle was confirmed by a quick “flick” of the mouse’s tail. Pain behavior testing was carried out before and 30 min after the intrathecal injection.
3
Preparation and Storage of PARP Inhibitors
4
Antagonism of AMPA and N-cadherin in Sucrose Preference
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The AMPA receptor antagonist CNQX, the selective AMPAR agonist CX546, and the N-cadherin antagonist ADH-1 were obtained from Sigma-Aldrich, St. Louis, MI, USA. CNQX and ADH-1 were dissolved in 0.9% saline solution, and CX546 was dissolved in 1% DMSO, 1% ethanol, and 98% saline. The rats were intraperitoneally injected with CNQX or ADH-1 (10 mg/kg) or an equal volume of saline and were subjected to sucrose preference test (SPT) 0.5 h after administration [34, 35] . The CX546 doses used were the same as previously reported [36] , and drugs or the same volume of vector (1% DMSO, 1% ethanol, and 98% saline) were microinjected into the hippocampus 30 min before the SPT.
For the second experiment, female offspring rats were divided into 6 groups and 6 litters were used in each group: CON + saline (n = 6), PS + saline (n = 6), CON + CNQX (n = 6), PS + CNQX (n = 6), CON + ADH-1(n = 6), and PS + ADH-1 (n = 6); to avoid a "litter effect," 2 female pups from each litter at most were used in 5-7-weekold offspring. For the third experiment, male offspring rats were divided into 4 groups: CON + vector (n = 6), CON + CX546 (n = 6), PS + vector (n = 6), and PS + CX546 (n = 6), and at most 2 male pups from each litter were used in 5-week-old offspring.
For the second experiment, female offspring rats were divided into 6 groups and 6 litters were used in each group: CON + saline (n = 6), PS + saline (n = 6), CON + CNQX (n = 6), PS + CNQX (n = 6), CON + ADH-1(n = 6), and PS + ADH-1 (n = 6); to avoid a "litter effect," 2 female pups from each litter at most were used in 5-7-weekold offspring. For the third experiment, male offspring rats were divided into 4 groups: CON + vector (n = 6), CON + CX546 (n = 6), PS + vector (n = 6), and PS + CX546 (n = 6), and at most 2 male pups from each litter were used in 5-week-old offspring.
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