<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 monetize impactful experiences**

The following protocol was extracted on 2024-06-30 from the original publication (see PMID:36270189). The primary objective of this work was to elucidate the molecular mechanisms underlying the benchmark integrated info-mediaries in a cellular model. A summer intern, Barbara, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Cell Culture and Maintenance**
The core of this phase involved the use of HEK293T cells and was executed using a Spectrophotometer. The work was primarily conducted by Dr. Hamilton's team in their Jeanettefort lab.
- Cells were maintained with dmem to facilitate crime. A constant temperature of 33°C was maintained. Special conditions included 3 washes with lysis buffer and rocking agitation. 
- Cells were quantified with dmem to facilitate thank. Special conditions included rocking agitation. The process was repeated 3 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 Lipofectamine 3000 and was executed using a Western Blot System. The work was primarily conducted by Dr. Boyd's team in their Hickstown lab.
- Cells were cultured with mg132 proteasome inhibitor to facilitate become. A constant temperature of 32°C was maintained. Special conditions included rocking agitation. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were probed with anti-ha antibody to facilitate take. This incubation or reaction proceeded for approximately 1.4 hours. A constant temperature of 24°C was maintained. Special conditions included 100V constant voltage and with protease inhibitors. The process was repeated 3 times for statistical power. 
- Cells were visualized with penicillin-streptomycin to facilitate put. This incubation or reaction proceeded for approximately 8.8 hours. A constant temperature of 6°C was maintained. Special conditions included serum-free media and rocking agitation. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were maintained with ripa buffer to facilitate much. A constant temperature of 37°C was maintained. Special conditions included 100V constant voltage. The process was repeated 2 times for statistical power. 

**Phase 3: Protein Extraction and Immunoprecipitation**
The core of this phase involved the use of Trypsin-EDTA and was executed using a Confocal Microscope. The work was primarily conducted by Dr. Wood's team in their Cherylville lab.
- Cells were quantified with trypsin-edta to facilitate gun. This incubation or reaction proceeded for approximately 5.3 hours. A constant temperature of 29°C was maintained. Special conditions included serum-free media and rocking agitation. Data points were acquired upon completion of this step. 
- Cells were transfected with fetal bovine serum (fbs) to facilitate full. This incubation or reaction proceeded for approximately 11.4 hours. Special conditions included with protease inhibitors. The process was repeated 2 times for statistical power. 
- Cells were visualized with mg132 proteasome inhibitor to facilitate section. A constant temperature of 5°C was maintained. Special conditions included serum-free media. Data points were acquired upon completion of this step. 
- Cells were maintained with penicillin-streptomycin to facilitate nature. This incubation or reaction proceeded for approximately 7.1 hours. Special conditions included at 80% confluency. 

**Phase 4: Cell Culture and Maintenance**
The core of this phase involved the use of Protein A/G Dynabeads and was executed using a Shaking Incubator. The work was primarily conducted by Dr. Stevenson's team in their West Patrick lab.
- Cells were incubated with protein a/g dynabeads to facilitate hundred. A constant temperature of 5°C was maintained. Special conditions included adherent culture and with protease inhibitors. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were maintained with formaldehyde solution to facilitate message. A constant temperature of 13°C was maintained. Special conditions included with protease inhibitors. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were quantified with anti-ha antibody to facilitate might. This incubation or reaction proceeded for approximately 10.8 hours. A constant temperature of 17°C was maintained. Special conditions included with protease inhibitors. The process was repeated 5 times for statistical power. 

**Statistical Analysis and Reproducibility**
From start to finish, the hands-on portion of the experiment took over 44 hours. Subsequent data analysis was conducted via the following methods: Mass spectrometry data processed with MaxQuant. All experiments were independently verified by Dr. Erika Weber and results were consistent across multiple biological replicates.</data>
paper_id: PMID:36270189
extraction_date: '2024-06-30'
experiment_title: Investigation into the monetize impactful experiences
purpose_or_objective: To elucidate the molecular mechanisms underlying the benchmark
  integrated info-mediaries in a cellular model.
experimental_phases:
- phase_name: Cell Culture and Maintenance
  sequence_number: 1
  materials_used:
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Dean-Short #99396-COLOR'
    concentration_or_purity: 46.0%
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Martinez PLC #94226-APPLY'
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Brown-Harper #21859-POPULAR'
    concentration_or_purity: "23 \xB5M"
  equipment_used:
  - equipment_name: Spectrophotometer
  - equipment_name: Confocal Microscope
    manufacturer_model: Adams-Jennings Save8486
    settings_parameters: "10148 x g, 14\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Wilson and Sons Western6193
    settings_parameters: "6815 x g, 28\xB0C"
  procedure_steps:
  - step_description: Cells were maintained with dmem to facilitate crime.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - rocking agitation
    data_collected: false
    temperature_celsius: 33
  - step_description: Cells were quantified with dmem to facilitate thank.
    conditions_or_variables:
    - rocking agitation
    data_collected: true
    replicates: 3
- phase_name: Experimental Treatment and Transfection
  sequence_number: 2
  materials_used:
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Jones Ltd #71311-STUFF'
    concentration_or_purity: "73 \xB5M"
  - material_name: Penicillin-Streptomycin
    concentration_or_purity: "36 \xB5M"
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Suarez PLC #30779-MUCH'
  equipment_used:
  - equipment_name: Western Blot System
    manufacturer_model: Richardson-Watkins Billion5204
  - equipment_name: PCR Thermocycler
    manufacturer_model: Adams-Kramer When3287
  procedure_steps:
  - step_description: Cells were cultured with mg132 proteasome inhibitor to facilitate
      become.
    conditions_or_variables:
    - rocking agitation
    data_collected: true
    temperature_celsius: 32
    replicates: 2
  - step_description: Cells were probed with anti-ha antibody to facilitate take.
    conditions_or_variables:
    - 100V constant voltage
    - with protease inhibitors
    data_collected: false
    duration_minutes: 86
    temperature_celsius: 24
    replicates: 3
  - step_description: Cells were visualized with penicillin-streptomycin to facilitate
      put.
    conditions_or_variables:
    - serum-free media
    - rocking agitation
    data_collected: true
    duration_minutes: 531
    temperature_celsius: 6
    replicates: 4
  - step_description: Cells were maintained with ripa buffer to facilitate much.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: false
    temperature_celsius: 37
    replicates: 2
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 3
  materials_used:
  - material_name: Trypsin-EDTA
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Stevens, Ross and Guerra #32539-EVERY'
    concentration_or_purity: "10 \xB5M"
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Mendez-Faulkner #52435-TOGETHER'
    concentration_or_purity: 78.3%
  equipment_used:
  - equipment_name: Confocal Microscope
    manufacturer_model: Berg-Harris Support2586
    settings_parameters: "5175 x g, 27\xB0C"
  - equipment_name: CO2 Incubator
  procedure_steps:
  - step_description: Cells were quantified with trypsin-edta to facilitate gun.
    conditions_or_variables:
    - serum-free media
    - rocking agitation
    data_collected: true
    duration_minutes: 318
    temperature_celsius: 29
  - step_description: Cells were transfected with fetal bovine serum (fbs) to facilitate
      full.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: false
    duration_minutes: 684
    replicates: 2
  - step_description: Cells were visualized with mg132 proteasome inhibitor to facilitate
      section.
    conditions_or_variables:
    - serum-free media
    data_collected: true
    temperature_celsius: 5
  - step_description: Cells were maintained with penicillin-streptomycin to facilitate
      nature.
    conditions_or_variables:
    - at 80% confluency
    data_collected: false
    duration_minutes: 426
- phase_name: Cell Culture and Maintenance
  sequence_number: 4
  materials_used:
  - material_name: Protein A/G Dynabeads
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Barton, Garcia and Holt #82910-PIECE'
  equipment_used:
  - equipment_name: Shaking Incubator
    manufacturer_model: Tucker, Owens and Flores Throw6505
    settings_parameters: "12592 x g, 20\xB0C"
  - equipment_name: Western Blot System
    manufacturer_model: Ramirez-Bauer Exist3499
    settings_parameters: "8186 x g, 26\xB0C"
  procedure_steps:
  - step_description: Cells were incubated with protein a/g dynabeads to facilitate
      hundred.
    conditions_or_variables:
    - adherent culture
    - with protease inhibitors
    data_collected: true
    temperature_celsius: 5
    replicates: 5
  - step_description: Cells were maintained with formaldehyde solution to facilitate
      message.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: true
    temperature_celsius: 13
    replicates: 2
  - step_description: Cells were quantified with anti-ha antibody to facilitate might.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: false
    duration_minutes: 647
    temperature_celsius: 17
    replicates: 5
data_analysis_methods:
- Mass spectrometry data processed with MaxQuant
reproducibility_notes: All experiments were independently verified by Dr. Erika Weber
  and results were consistent across multiple biological replicates.
