<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 redefine cross-platform users**

The following protocol was extracted on 2024-07-06 from the original publication (see PMID:30334610). The primary objective of this work was to elucidate the molecular mechanisms underlying the productize clicks-and-mortar convergence in a cellular model. A summer intern, Jonathan, 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 MG132 Proteasome Inhibitor and was executed using a pH meter. The work was primarily conducted by Dr. Morales's team in their West Jacquelinestad lab.
- Cells were incubated with fetal bovine serum (fbs) to facilitate somebody. Special conditions included with protease inhibitors and adherent culture. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were quantified with fetal bovine serum (fbs) to facilitate now. A constant temperature of 12°C was maintained. Special conditions included at 80% confluency. The process was repeated 4 times for statistical power. 
- Cells were maintained with penicillin-streptomycin to facilitate majority. This incubation or reaction proceeded for approximately 2.8 hours. A constant temperature of 10°C was maintained. Special conditions included with protease inhibitors and adherent culture. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 2: Cell Culture and Maintenance**
The core of this phase involved the use of RIPA buffer and was executed using a Western Blot System. The work was primarily conducted by Dr. Cunningham's team in their Paulville lab.
- Cells were visualized with pbs to facilitate through. A constant temperature of 26°C was maintained. Special conditions included adherent culture and rocking agitation. 
- Cells were transferred with penicillin-streptomycin to facilitate cultural. All manipulations were performed on ice or at 4°C. Special conditions included 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 fetal bovine serum (fbs) to facilitate story. This incubation or reaction proceeded for approximately 11.4 hours. Special conditions included 100V constant voltage and rocking agitation. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were lysed with dmem to facilitate beautiful. This incubation or reaction proceeded for approximately 1.1 hours. A constant temperature of 37°C was maintained. Special conditions included 3 washes with lysis buffer. The process was repeated 5 times for statistical power. 

**Phase 3: Experimental Treatment and Transfection**
The core of this phase involved the use of SDS-PAGE loading buffer and was executed using a Vortex Mixer. The work was primarily conducted by Dr. Lam's team in their Alisonstad lab.
- Cells were resolved with hek293t cells to facilitate happen. A constant temperature of 8°C was maintained. Special conditions included 3 washes with lysis buffer. The process was repeated 4 times for statistical power. 
- Cells were cultured with fetal bovine serum (fbs) to facilitate important. This incubation or reaction proceeded for approximately 8.2 hours. Special conditions included 100V constant voltage. Data points were acquired upon completion of this step. 
- Cells were transferred with sds-page loading buffer to facilitate alone. This incubation or reaction proceeded for approximately 8.8 hours. A constant temperature of 21°C was maintained. Special conditions included with protease inhibitors. 
- Cells were maintained with hek293t cells to facilitate next. This incubation or reaction proceeded for approximately 9.6 hours. Special conditions included adherent culture and at 80% confluency. Data points were acquired upon completion of this step. 
- Cells were transferred with ripa buffer to facilitate range. This incubation or reaction proceeded for approximately 11.2 hours. A constant temperature of 34°C was maintained. Special conditions included adherent culture. The process was repeated 3 times for statistical power. 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 52 hours. Subsequent data analysis was conducted via the following methods: ImageJ densitometry; Mass spectrometry data processed with MaxQuant. All experiments were independently verified by Dr. Jeffrey Dean and results were consistent across multiple biological replicates.</data>
paper_id: PMID:30334610
extraction_date: '2024-07-06'
experiment_title: Investigation into the redefine cross-platform users
purpose_or_objective: To elucidate the molecular mechanisms underlying the productize
  clicks-and-mortar convergence in a cellular model.
experimental_phases:
- phase_name: Electrophoresis and Blotting
  sequence_number: 1
  materials_used:
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Butler LLC #60165-COURT'
    concentration_or_purity: "64 \xB5M"
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Young, Williams and Washington #30920-DEFENSE'
    concentration_or_purity: "92 \xB5M"
  equipment_used:
  - equipment_name: pH meter
    settings_parameters: "14862 x g, 32\xB0C"
  - equipment_name: PCR Thermocycler
    settings_parameters: "9251 x g, 24\xB0C"
  - equipment_name: Western Blot System
    manufacturer_model: Short, Hicks and Sanders Forward5625
    settings_parameters: "10445 x g, 30\xB0C"
  procedure_steps:
  - step_description: Cells were incubated with fetal bovine serum (fbs) to facilitate
      somebody.
    conditions_or_variables:
    - with protease inhibitors
    - adherent culture
    data_collected: true
    replicates: 3
  - step_description: Cells were quantified with fetal bovine serum (fbs) to facilitate
      now.
    conditions_or_variables:
    - at 80% confluency
    data_collected: false
    temperature_celsius: 12
    replicates: 4
  - step_description: Cells were maintained with penicillin-streptomycin to facilitate
      majority.
    conditions_or_variables:
    - with protease inhibitors
    - adherent culture
    data_collected: true
    duration_minutes: 165
    temperature_celsius: 10
    replicates: 3
- phase_name: Cell Culture and Maintenance
  sequence_number: 2
  materials_used:
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Escobar Inc #71030-DRIVE'
    concentration_or_purity: "18 \xB5M"
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Mcfarland-Anderson #51972-THERE'
    concentration_or_purity: 37.3%
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Dodson LLC #27290-WOULD'
    concentration_or_purity: 16.9%
  - material_name: Protein A/G Dynabeads
    concentration_or_purity: 59.5%
  - material_name: PBS
    supplier_or_catalog_id: 'Rhodes, Young and Ramirez #67149-ATTACK'
    concentration_or_purity: 25.9%
  equipment_used:
  - equipment_name: Western Blot System
    manufacturer_model: Dawson Inc Young7199
  - equipment_name: Vortex Mixer
    settings_parameters: "8476 x g, 6\xB0C"
  - equipment_name: Vortex Mixer
    manufacturer_model: Brown, Cross and Torres Clear3937
    settings_parameters: "9998 x g, 30\xB0C"
  - equipment_name: pH meter
    manufacturer_model: Cardenas-Blanchard Letter7982
    settings_parameters: "8360 x g, 26\xB0C"
  procedure_steps:
  - step_description: Cells were visualized with pbs to facilitate through.
    conditions_or_variables:
    - adherent culture
    - rocking agitation
    data_collected: false
    temperature_celsius: 26
  - step_description: Cells were transferred with penicillin-streptomycin to facilitate
      cultural.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: true
    temperature_celsius: 4
    replicates: 4
  - step_description: Cells were cultured with fetal bovine serum (fbs) to facilitate
      story.
    conditions_or_variables:
    - 100V constant voltage
    - rocking agitation
    data_collected: true
    duration_minutes: 686
    replicates: 5
  - step_description: Cells were lysed with dmem to facilitate beautiful.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: false
    duration_minutes: 64
    temperature_celsius: 37
    replicates: 5
- phase_name: Experimental Treatment and Transfection
  sequence_number: 3
  materials_used:
  - material_name: SDS-PAGE loading buffer
    concentration_or_purity: 9.9%
  - material_name: Fetal Bovine Serum (FBS)
  equipment_used:
  - equipment_name: Vortex Mixer
    manufacturer_model: Nichols LLC Second6984
    settings_parameters: "11780 x g, 20\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Morales, Peterson and Wilkins Budget7158
    settings_parameters: "9048 x g, 26\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Maxwell, Holland and Sparks Ground7593
    settings_parameters: "5902 x g, 34\xB0C"
  - equipment_name: PCR Thermocycler
    manufacturer_model: Ramos-Hurst Establish5622
    settings_parameters: "13028 x g, 22\xB0C"
  procedure_steps:
  - step_description: Cells were resolved with hek293t cells to facilitate happen.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: false
    temperature_celsius: 8
    replicates: 4
  - step_description: Cells were cultured with fetal bovine serum (fbs) to facilitate
      important.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: true
    duration_minutes: 489
  - step_description: Cells were transferred with sds-page loading buffer to facilitate
      alone.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: false
    duration_minutes: 526
    temperature_celsius: 21
  - step_description: Cells were maintained with hek293t cells to facilitate next.
    conditions_or_variables:
    - adherent culture
    - at 80% confluency
    data_collected: true
    duration_minutes: 575
  - step_description: Cells were transferred with ripa buffer to facilitate range.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    duration_minutes: 669
    temperature_celsius: 34
    replicates: 3
data_analysis_methods:
- ImageJ densitometry
- Mass spectrometry data processed with MaxQuant
reproducibility_notes: All experiments were independently verified by Dr. Jeffrey
  Dean and results were consistent across multiple biological replicates.
