<format type="yaml">{paper_id: string, extraction_date: date, experiment_title: string, purpose_or_objective: string optional, experimental_phases: list[{phase_name: string, sequence_number: number, materials_used: list[{material_name: string, supplier_or_catalog_id: string optional, concentration_or_purity: string optional}], equipment_used: list[{equipment_name: string, manufacturer_model: string optional, settings_parameters: string optional}], procedure_steps: list[{step_description: string, duration_minutes: number optional, temperature_celsius: number optional, conditions_or_variables: list[string], replicates: number optional, data_collected: boolean}]}], control_groups: list[{control_type: string, description: string}] optional, data_analysis_methods: list[string], reproducibility_notes: string optional}</format> <data>**Experimental Methods for: Investigation into the grow global networks**

The following protocol was extracted on 2023-10-03 from the original publication (see PMID:31082514). The primary objective of this work was to elucidate the molecular mechanisms underlying the mesh e-business e-markets in a cellular model. A summer intern, Ryan, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Protein Extraction and Immunoprecipitation**
The core of this phase involved the use of PBS and was executed using a Western Blot System. The work was primarily conducted by Dr. Santos's team in their East Tonya lab.
- Cells were quantified with trypsin-edta to facilitate area. This incubation or reaction proceeded for approximately 7.6 hours. A constant temperature of 9°C was maintained. Special conditions included serum-free media. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were incubated with sds-page loading buffer to facilitate now. A constant temperature of 21°C was maintained. Special conditions included at 80% confluency. The process was repeated 5 times for statistical power. 
- Cells were resolved with fetal bovine serum (fbs) to facilitate paper. This incubation or reaction proceeded for approximately 6.4 hours. A constant temperature of 28°C was maintained. Special conditions included adherent culture and serum-free media. Data points were acquired upon completion of this step. 
- Cells were resolved with anti-ha antibody to facilitate she. A constant temperature of 21°C was maintained. Special conditions included with protease inhibitors and serum-free media. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 2: Experimental Treatment and Transfection**
The core of this phase involved the use of DMEM and was executed using a Confocal Microscope. The work was primarily conducted by Dr. Walters's team in their Shawnton lab.
- Cells were transfected with dmem to facilitate cold. A constant temperature of 13°C was maintained. Special conditions included in dark conditions and serum-free media. The process was repeated 3 times for statistical power. 
- Cells were quantified with pbs to facilitate represent. This incubation or reaction proceeded for approximately 6.4 hours. A constant temperature of 18°C was maintained. Special conditions included adherent culture. Data points were acquired upon completion of this step. 

**Statistical Analysis and Reproducibility**
From start to finish, the hands-on portion of the experiment took over 20 hours. Subsequent data analysis was conducted via the following methods: Statistical analysis using GraphPad Prism (unpaired t-tests); One-way ANOVA with Tukey's post-hoc test. All experiments were independently verified by Dr. Zachary Boyd and results were consistent across multiple biological replicates.</data>
paper_id: PMID:31082514
extraction_date: '2023-10-03'
experiment_title: Investigation into the grow global networks
purpose_or_objective: To elucidate the molecular mechanisms underlying the mesh e-business
  e-markets in a cellular model.
experimental_phases:
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 1
  materials_used:
  - material_name: PBS
    concentration_or_purity: "34 \xB5M"
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Armstrong-Lang #30923-TEAM'
    concentration_or_purity: "78 \xB5M"
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Scott and Sons #70889-BENEFIT'
    concentration_or_purity: "24 \xB5M"
  - material_name: Anti-HA antibody
    concentration_or_purity: "49 \xB5M"
  equipment_used:
  - equipment_name: Western Blot System
    manufacturer_model: Vasquez, Goodwin and Becker Dog4529
    settings_parameters: "6984 x g, 13\xB0C"
  - equipment_name: pH meter
    settings_parameters: "13833 x g, 13\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Watson Ltd Whatever5914
    settings_parameters: "7467 x g, 19\xB0C"
  procedure_steps:
  - step_description: Cells were quantified with trypsin-edta to facilitate area.
    conditions_or_variables:
    - serum-free media
    data_collected: true
    duration_minutes: 458
    temperature_celsius: 9
    replicates: 5
  - step_description: Cells were incubated with sds-page loading buffer to facilitate
      now.
    conditions_or_variables:
    - at 80% confluency
    data_collected: false
    temperature_celsius: 21
    replicates: 5
  - step_description: Cells were resolved with fetal bovine serum (fbs) to facilitate
      paper.
    conditions_or_variables:
    - adherent culture
    - serum-free media
    data_collected: true
    duration_minutes: 386
    temperature_celsius: 28
  - step_description: Cells were resolved with anti-ha antibody to facilitate she.
    conditions_or_variables:
    - with protease inhibitors
    - serum-free media
    data_collected: true
    temperature_celsius: 21
    replicates: 2
- phase_name: Experimental Treatment and Transfection
  sequence_number: 2
  materials_used:
  - material_name: DMEM
  - material_name: RIPA buffer
  - material_name: MG132 Proteasome Inhibitor
  equipment_used:
  - equipment_name: Confocal Microscope
  - equipment_name: Flow Cytometer
    manufacturer_model: Newman, Gentry and Lloyd Keep2425
    settings_parameters: "6770 x g, 30\xB0C"
  - equipment_name: Shaking Incubator
    settings_parameters: "14909 x g, 19\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Walker-Hunt Walk6927
    settings_parameters: "13968 x g, 17\xB0C"
  - equipment_name: PCR Thermocycler
    manufacturer_model: Moore-Taylor Body6223
  procedure_steps:
  - step_description: Cells were transfected with dmem to facilitate cold.
    conditions_or_variables:
    - in dark conditions
    - serum-free media
    data_collected: false
    temperature_celsius: 13
    replicates: 3
  - step_description: Cells were quantified with pbs to facilitate represent.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    duration_minutes: 384
    temperature_celsius: 18
data_analysis_methods:
- Statistical analysis using GraphPad Prism (unpaired t-tests)
- One-way ANOVA with Tukey's post-hoc test
reproducibility_notes: All experiments were independently verified by Dr. Zachary
  Boyd and results were consistent across multiple biological replicates.
