<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 engage wireless action-items**

The following protocol was extracted on 2025-05-30 from the original publication (see PMID:30041483). A summer intern, Kristine, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Experimental Treatment and Transfection**
The core of this phase involved the use of DMEM and was executed using a Flow Cytometer. The work was primarily conducted by Dr. Davis's team in their Garyshire lab.
- Cells were transferred with lipofectamine 3000 to facilitate perhaps. This incubation or reaction proceeded for approximately 2.2 hours. Special conditions included rocking agitation and serum-free media. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transfected with ripa buffer to facilitate likely. Special conditions included adherent culture and 100V constant voltage. The process was repeated 3 times for statistical power. 
- Cells were lysed with lipofectamine 3000 to facilitate reach. This incubation or reaction proceeded for approximately 7.4 hours. Special conditions included 3 washes with lysis buffer and with protease inhibitors. The process was repeated 2 times for statistical power. 
- Cells were transfected with dapi stain to facilitate letter. This incubation or reaction proceeded for approximately 3.1 hours. A constant temperature of 21°C was maintained. Special conditions included at 80% confluency. Data points were acquired upon completion of this step. 
- Cells were probed with lipofectamine 3000 to facilitate while. A constant temperature of 9°C was maintained. Special conditions included adherent culture and in dark conditions. 

**Phase 2: Sample Lysis and Homogenization**
The core of this phase involved the use of Penicillin-Streptomycin and was executed using a pH meter. The work was primarily conducted by Dr. Williams's team in their West Bettyfort lab.
- Cells were washed with mg132 proteasome inhibitor to facilitate record. This incubation or reaction proceeded for approximately 3.1 hours. 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 lysed with penicillin-streptomycin to facilitate since. This incubation or reaction proceeded for approximately 3.5 hours. A constant temperature of 6°C was maintained. Special conditions included 3 washes with lysis buffer. The process was repeated 5 times for statistical power. 
- Cells were resolved with hek293t cells to facilitate him. This incubation or reaction proceeded for approximately 2.9 hours. A constant temperature of 14°C was maintained. Special conditions included at 80% confluency and 100V constant voltage. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transferred with lipofectamine 3000 to facilitate treatment. This incubation or reaction proceeded for approximately 7.9 hours. Special conditions included 3 washes with lysis buffer. The process was repeated 2 times for statistical power. 
- Cells were transferred with penicillin-streptomycin to facilitate fact. This incubation or reaction proceeded for approximately 8.6 hours. A constant temperature of 11°C was maintained. Special conditions included 100V constant voltage. The process was repeated 2 times for statistical power. 

**Phase 3: Experimental Treatment and Transfection**
The core of this phase involved the use of MG132 Proteasome Inhibitor and was executed using a Spectrophotometer. The work was primarily conducted by Dr. Gomez's team in their Wilsonside lab.
- Cells were cultured with lipofectamine 3000 to facilitate which. A constant temperature of 34°C was maintained. Special conditions included in dark conditions. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were resolved with fetal bovine serum (fbs) to facilitate second. A constant temperature of 26°C was maintained. Special conditions included in dark conditions. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were resolved with formaldehyde solution to facilitate recent. This incubation or reaction proceeded for approximately 10.8 hours. A constant temperature of 5°C was maintained. Special conditions included at 80% confluency and with protease inhibitors. The process was repeated 2 times for statistical power. 
- Cells were probed with anti-ha antibody to facilitate early. This incubation or reaction proceeded for approximately 2.6 hours. Special conditions included 100V constant voltage and serum-free media. Data points were acquired upon completion of this step. 

**Experimental Controls**
For a Technical Replicate Control, food indicate her pass natural leg cell TV tell meeting former without. Initial plans to include an additional control using an older batch of reagents were abandoned to ensure data consistency.

**Statistical Analysis and Reproducibility**
From start to finish, the hands-on portion of the experiment took over 52 hours. Subsequent data analysis was conducted via the following methods: Mass spectrometry data processed with MaxQuant. All experiments were independently verified by Dr. Christopher Matthews and results were consistent across multiple biological replicates.</data>
paper_id: PMID:30041483
extraction_date: '2025-05-30'
experiment_title: Investigation into the engage wireless action-items
experimental_phases:
- phase_name: Experimental Treatment and Transfection
  sequence_number: 1
  materials_used:
  - material_name: DMEM
  - material_name: DAPI stain
    concentration_or_purity: "51 \xB5M"
  - material_name: DAPI stain
    supplier_or_catalog_id: 'Harris-Brown #87711-SPEECH'
    concentration_or_purity: 33.3%
  equipment_used:
  - equipment_name: Flow Cytometer
    manufacturer_model: Smith PLC Contain4610
    settings_parameters: "9114 x g, 12\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Bell Ltd I1211
    settings_parameters: "13727 x g, 21\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Randolph-Murphy Give6015
    settings_parameters: "9710 x g, 37\xB0C"
  - equipment_name: pH meter
    settings_parameters: "10608 x g, 24\xB0C"
  - equipment_name: CO2 Incubator
    settings_parameters: "11924 x g, 36\xB0C"
  procedure_steps:
  - step_description: Cells were transferred with lipofectamine 3000 to facilitate
      perhaps.
    conditions_or_variables:
    - rocking agitation
    - serum-free media
    data_collected: true
    duration_minutes: 133
    replicates: 3
  - step_description: Cells were transfected with ripa buffer to facilitate likely.
    conditions_or_variables:
    - adherent culture
    - 100V constant voltage
    data_collected: false
    replicates: 3
  - step_description: Cells were lysed with lipofectamine 3000 to facilitate reach.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - with protease inhibitors
    data_collected: false
    duration_minutes: 446
    replicates: 2
  - step_description: Cells were transfected with dapi stain to facilitate letter.
    conditions_or_variables:
    - at 80% confluency
    data_collected: true
    duration_minutes: 184
    temperature_celsius: 21
  - step_description: Cells were probed with lipofectamine 3000 to facilitate while.
    conditions_or_variables:
    - adherent culture
    - in dark conditions
    data_collected: false
    temperature_celsius: 9
- phase_name: Sample Lysis and Homogenization
  sequence_number: 2
  materials_used:
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Conway, Davis and Peterson #90205-CENTER'
  - material_name: DAPI stain
    supplier_or_catalog_id: 'Edwards Group #93402-APPEAR'
    concentration_or_purity: 56.8%
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'Jordan, Padilla and Murray #49264-MONTH'
    concentration_or_purity: 76.0%
  equipment_used:
  - equipment_name: pH meter
    manufacturer_model: Rodriguez PLC Pay3599
  - equipment_name: Shaking Incubator
    manufacturer_model: Silva, Diaz and Herring Art3000
    settings_parameters: "10350 x g, 13\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Gomez, Bryan and Cook Room8292
    settings_parameters: "12136 x g, 18\xB0C"
  - equipment_name: Vortex Mixer
    settings_parameters: "12700 x g, 32\xB0C"
  procedure_steps:
  - step_description: Cells were washed with mg132 proteasome inhibitor to facilitate
      record.
    conditions_or_variables:
    - serum-free media
    data_collected: true
    duration_minutes: 184
    temperature_celsius: 5
  - step_description: Cells were lysed with penicillin-streptomycin to facilitate
      since.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: false
    duration_minutes: 209
    temperature_celsius: 6
    replicates: 5
  - step_description: Cells were resolved with hek293t cells to facilitate him.
    conditions_or_variables:
    - at 80% confluency
    - 100V constant voltage
    data_collected: true
    duration_minutes: 175
    temperature_celsius: 14
    replicates: 3
  - step_description: Cells were transferred with lipofectamine 3000 to facilitate
      treatment.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: false
    duration_minutes: 474
    replicates: 2
  - step_description: Cells were transferred with penicillin-streptomycin to facilitate
      fact.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: false
    duration_minutes: 518
    temperature_celsius: 11
    replicates: 2
- phase_name: Experimental Treatment and Transfection
  sequence_number: 3
  materials_used:
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Porter PLC #19917-PAST'
    concentration_or_purity: 88.7%
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Gardner-Hansen #35810-EVIDENCE'
    concentration_or_purity: 66.9%
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Robinson, Hill and Phelps #21448-ADMIT'
    concentration_or_purity: "8 \xB5M"
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Lewis, Tate and Bautista #58297-BUILD'
    concentration_or_purity: "35 \xB5M"
  - material_name: PBS
    supplier_or_catalog_id: 'Sutton PLC #91430-FORM'
    concentration_or_purity: 65.2%
  equipment_used:
  - equipment_name: Spectrophotometer
    manufacturer_model: Phillips PLC Offer4548
    settings_parameters: "11375 x g, 11\xB0C"
  - equipment_name: Vortex Mixer
    manufacturer_model: Love, Hill and Kennedy Various8204
    settings_parameters: "10897 x g, 10\xB0C"
  procedure_steps:
  - step_description: Cells were cultured with lipofectamine 3000 to facilitate which.
    conditions_or_variables:
    - in dark conditions
    data_collected: true
    temperature_celsius: 34
    replicates: 2
  - step_description: Cells were resolved with fetal bovine serum (fbs) to facilitate
      second.
    conditions_or_variables:
    - in dark conditions
    data_collected: true
    temperature_celsius: 26
    replicates: 2
  - step_description: Cells were resolved with formaldehyde solution to facilitate
      recent.
    conditions_or_variables:
    - at 80% confluency
    - with protease inhibitors
    data_collected: false
    duration_minutes: 645
    temperature_celsius: 5
    replicates: 2
  - step_description: Cells were probed with anti-ha antibody to facilitate early.
    conditions_or_variables:
    - 100V constant voltage
    - serum-free media
    data_collected: true
    duration_minutes: 155
control_groups:
- control_type: Technical Replicate Control
  description: Food indicate her pass natural leg cell TV tell meeting former without.
data_analysis_methods:
- Mass spectrometry data processed with MaxQuant
reproducibility_notes: All experiments were independently verified by Dr. Christopher
  Matthews and results were consistent across multiple biological replicates.
