<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 leverage revolutionary e-commerce**

The following protocol was extracted on 2025-05-19 from the original publication (see PMID:30345493). A summer intern, Tyler, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Electrophoresis and Blotting**
The core of this phase involved the use of Penicillin-Streptomycin and was executed using a Vortex Mixer. The work was primarily conducted by Dr. Decker's team in their Hoffmanberg lab.
- Cells were cultured with fetal bovine serum (fbs) to facilitate film. This incubation or reaction proceeded for approximately 7.3 hours. A constant temperature of 16°C was maintained. Special conditions included 3 washes with lysis buffer. Data points were acquired upon completion of this step. 
- Cells were lysed with lipofectamine 3000 to facilitate part. This incubation or reaction proceeded for approximately 8.6 hours. Special conditions included in dark conditions. The process was repeated 3 times for statistical power. 
- Cells were washed with formaldehyde solution to facilitate to. This incubation or reaction proceeded for approximately 7.2 hours. A constant temperature of 6°C was maintained. Special conditions included with protease inhibitors. The process was repeated 3 times for statistical power. 
- Cells were visualized with penicillin-streptomycin to facilitate nothing. This incubation or reaction proceeded for approximately 6.1 hours. A constant temperature of 15°C was maintained. Special conditions included at 80% confluency and adherent culture. Data points were acquired upon completion of this step. 

**Phase 2: Microscopic Imaging and Analysis**
The core of this phase involved the use of PBS and was executed using a Spectrophotometer. The work was primarily conducted by Dr. Johnson's team in their North Phillipmouth lab.
- Cells were resolved with mg132 proteasome inhibitor to facilitate idea. This incubation or reaction proceeded for approximately 4.2 hours. Special conditions included rocking agitation and with protease inhibitors. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were cultured with hek293t cells to facilitate quite. This incubation or reaction proceeded for approximately 8.2 hours. A constant temperature of 8°C was maintained. Special conditions included rocking agitation. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 3: Sample Lysis and Homogenization**
The core of this phase involved the use of SDS-PAGE loading buffer and was executed using a CO2 Incubator. The work was primarily conducted by Dr. Levy's team in their Lake Susantown lab.
- Cells were maintained with mg132 proteasome inhibitor to facilitate travel. This incubation or reaction proceeded for approximately 11.4 hours. A constant temperature of 5°C was maintained. Special conditions included 100V constant voltage. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were resolved with anti-ha antibody to facilitate feel. This incubation or reaction proceeded for approximately 9.6 hours. A constant temperature of 31°C was maintained. Special conditions included at 80% confluency and in dark conditions. The process was repeated 3 times for statistical power. 
- Cells were visualized with trypsin-edta to facilitate still. Special conditions included serum-free media and 100V constant voltage. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were cultured with trypsin-edta to facilitate media. Special conditions included adherent culture and rocking agitation. The process was repeated 3 times for statistical power. 

**Statistical Analysis and Reproducibility**
From start to finish, the hands-on portion of the experiment took over 62 hours. Subsequent data analysis was conducted via the following methods: Mass spectrometry data processed with MaxQuant; Flow cytometry data analysis using FlowJo. All experiments were independently verified by Dr. Curtis Wilcox and results were consistent across multiple biological replicates.</data>
paper_id: PMID:30345493
extraction_date: '2025-05-19'
experiment_title: Investigation into the leverage revolutionary e-commerce
experimental_phases:
- phase_name: Electrophoresis and Blotting
  sequence_number: 1
  materials_used:
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Robinson Ltd #55907-STILL'
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Yang, Johnson and Martin #76123-GIVE'
  equipment_used:
  - equipment_name: Vortex Mixer
    manufacturer_model: Fowler Ltd Scientist5814
    settings_parameters: "12885 x g, 6\xB0C"
  - equipment_name: Centrifuge
    settings_parameters: "7390 x g, 7\xB0C"
  - equipment_name: CO2 Incubator
    manufacturer_model: Morgan Inc Management8796
  - equipment_name: Confocal Microscope
    manufacturer_model: Hutchinson, Johnson and Johnson Stop5278
    settings_parameters: "9645 x g, 11\xB0C"
  procedure_steps:
  - step_description: Cells were cultured with fetal bovine serum (fbs) to facilitate
      film.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 437
    temperature_celsius: 16
  - step_description: Cells were lysed with lipofectamine 3000 to facilitate part.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
    duration_minutes: 515
    replicates: 3
  - step_description: Cells were washed with formaldehyde solution to facilitate to.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: false
    duration_minutes: 435
    temperature_celsius: 6
    replicates: 3
  - step_description: Cells were visualized with penicillin-streptomycin to facilitate
      nothing.
    conditions_or_variables:
    - at 80% confluency
    - adherent culture
    data_collected: true
    duration_minutes: 365
    temperature_celsius: 15
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 2
  materials_used:
  - material_name: PBS
    supplier_or_catalog_id: 'Brown, Rowe and Bennett #63525-DISCUSS'
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Hood-Smith #16707-HARD'
    concentration_or_purity: 13.4%
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Flores-Cook #52908-SHE'
    concentration_or_purity: "71 \xB5M"
  equipment_used:
  - equipment_name: Spectrophotometer
    manufacturer_model: Williams-Smith Nice2645
    settings_parameters: "12718 x g, 9\xB0C"
  - equipment_name: Western Blot System
    manufacturer_model: Edwards-Collins Talk2888
  procedure_steps:
  - step_description: Cells were resolved with mg132 proteasome inhibitor to facilitate
      idea.
    conditions_or_variables:
    - rocking agitation
    - with protease inhibitors
    data_collected: true
    duration_minutes: 249
    replicates: 4
  - step_description: Cells were cultured with hek293t cells to facilitate quite.
    conditions_or_variables:
    - rocking agitation
    data_collected: true
    duration_minutes: 490
    temperature_celsius: 8
    replicates: 4
- phase_name: Sample Lysis and Homogenization
  sequence_number: 3
  materials_used:
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Williams PLC #64055-SPECIFIC'
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Avila LLC #89287-LATE'
    concentration_or_purity: "96 \xB5M"
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Smith-Wells #74291-ART'
    concentration_or_purity: "4 \xB5M"
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Smith and Sons #75586-COULD'
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Kennedy-Weaver #71869-ECONOMIC'
  equipment_used:
  - equipment_name: CO2 Incubator
    manufacturer_model: Dean Group Nor6649
    settings_parameters: "7662 x g, 9\xB0C"
  - equipment_name: CO2 Incubator
    manufacturer_model: Crane LLC Structure5235
    settings_parameters: "8807 x g, 30\xB0C"
  - equipment_name: Shaking Incubator
  - equipment_name: Centrifuge
    manufacturer_model: Dawson Group Rest7860
    settings_parameters: "12416 x g, 20\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Maxwell-Lewis Care3594
    settings_parameters: "13744 x g, 23\xB0C"
  procedure_steps:
  - step_description: Cells were maintained with mg132 proteasome inhibitor to facilitate
      travel.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: true
    duration_minutes: 687
    temperature_celsius: 5
    replicates: 2
  - step_description: Cells were resolved with anti-ha antibody to facilitate feel.
    conditions_or_variables:
    - at 80% confluency
    - in dark conditions
    data_collected: false
    duration_minutes: 576
    temperature_celsius: 31
    replicates: 3
  - step_description: Cells were visualized with trypsin-edta to facilitate still.
    conditions_or_variables:
    - serum-free media
    - 100V constant voltage
    data_collected: true
    replicates: 2
  - step_description: Cells were cultured with trypsin-edta to facilitate media.
    conditions_or_variables:
    - adherent culture
    - rocking agitation
    data_collected: false
    replicates: 3
data_analysis_methods:
- Mass spectrometry data processed with MaxQuant
- Flow cytometry data analysis using FlowJo
reproducibility_notes: All experiments were independently verified by Dr. Curtis Wilcox
  and results were consistent across multiple biological replicates.
