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Nle8 18 tyr34 pth 1 34

Manufactured by Bachem

Nle8,18,Tyr34]-PTH(1-34) is a synthetic peptide compound used in scientific research and laboratory applications. It consists of a sequence of 34 amino acids, with specific modifications at positions 8, 18, and 34. This product is typically utilized as a tool for investigating and studying various biological processes and pathways.

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3 protocols using nle8 18 tyr34 pth 1 34

1

Oxidative Stress Modulation of PTHR Signaling

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HEK-293 cells were stably transfected with human influenza hemagglutinin (HA)-PTHR (HEK-293R) [14 (link)] and cultured in Dulbecco’s modified Eagle’s medium/F-12 medium supplemented with 10% fetal bovine serum, 100 units/ml penicillin, 100 μg/ml streptomycin, and 0.75 mg/ml geneticin. HEK-293R cells express 6.0 × 105 PTHR/cell, with an average Kd of 14.2 nM. Cells were maintained at 37 °C in a humidified atmosphere of 5% CO2, 95% air. Cells were pre-incubated for 15 min with different concentrations of H2O2 (1–500 μM) followed by stimulation with 10 nM–1 μM [Nle8,18,Tyr34]-PTH(1-34) (hereafter NNT-PTH) (Bachem, Torrance, CA), an oxidant-resistant PTH peptide.
For transient transfections, HEK-293R cells were grown on poly-D-lysine-coated glass 25-mm coverslips for 12 h prior to transfection with FuGENE 6 (Roche Applied Science) in complete medium. After 24 h, coverslips in HEPES/ bovine serum albumin buffer (pH=7.4) (HEPES buffer containing 0.1% (w/v) BSA) were transferred to an Attofluor chamber (Invitrogen, Carlsbad, CA) for live-cell imaging.
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2

Characterization of PTH1R Expression

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Lipofectamine 2000 was purchased from Thermo Fisher Scientific. [Nle8,18,Tyr34]PTH (1-34), a chemically more stable variant of native PTH, was purchased from Bachem and is referred to as PTH(1-34). Generation of a polyclonal rabbit anti-PTH1R (1781) antiserum was described previously (27 (link)). Anti-rabbit peroxidase-conjugated secondary antibodies were obtained from Dianova. Cy2-conjugated anti-rabbit antibody was from Jackson Immuno Research Lab. All cell culture media were obtained from PAN Biotech. All other reagents were of analytical grade from Sigma-Aldrich or Applichem.
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3

Antibody and Protein Source Verification

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Monoclonal anti-HA agarose (catalog no.: A2095) and rabbit polyclonal anti-FLAG (catalog no.: F7425) were purchased from MilliporeSigma. Rabbit polyclonal anti-HA (catalog no.: sc-805) and protein G+ agarose (catalog no.: sc-2002) were purchased from Santa Cruz Biotechnology. Mouse and rabbit polyclonal anti-NHERF1 antibodies were purchased from Abcam (catalog nos.: ab9526 and ab3452, respectively). Rabbit polyclonal anti-RGS14 was purchased from Proteintech (catalog no.: 16258-1-AP). We bought antihuman NPT2A from Novus Biologicals (catalog no.: NBP242216) and antiactin from Santa Cruz Biotechnology (catalog no.: sc-1616R). HA-NHERF1 as described (46 (link)). [Nle8,18,Tyr34]PTH(1–34) was from Bachem (catalog no.: H9110). Recombinant human Arg179Gln-FGF2325–251 (referred to as FGF23), which is resistant to furin cleavage and inactivation, was obtained from R&D Systems (catalog no.: 2604-FG-025). PF-06869206 (catalog no.: PZ0389) was purchased from MilliporeSigma.
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