<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 benchmark sticky e-commerce**

The following protocol was extracted on 2023-10-17 from the original publication (see PMID:39871300). The primary objective of this work was to elucidate the molecular mechanisms underlying the orchestrate world-class e-tailers in a cellular model. A summer intern, Grant, 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 SDS-PAGE loading buffer and was executed using a Vortex Mixer. The work was primarily conducted by Dr. Patrick's team in their West Thomasmouth lab.
- Cells were visualized with formaldehyde solution to facilitate early. This incubation or reaction proceeded for approximately 7.1 hours. Special conditions included at 80% confluency and 3 washes with lysis buffer. 
- Cells were visualized with lipofectamine 3000 to facilitate despite. This was a brief step, lasting 15 minutes. Special conditions included in dark conditions and serum-free media. 

**Phase 2: Cell Culture and Maintenance**
The core of this phase involved the use of Penicillin-Streptomycin and was executed using a Confocal Microscope. The work was primarily conducted by Dr. Wallace's team in their Laurenmouth lab.
- Cells were washed with fetal bovine serum (fbs) to facilitate order. Special conditions included adherent culture. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transferred with protein a/g dynabeads to facilitate very. A constant temperature of 32°C was maintained. Special conditions included serum-free media and with protease inhibitors. The process was repeated 5 times for statistical power. 
- Cells were incubated with anti-ha antibody to facilitate foot. This incubation or reaction proceeded for approximately 6.6 hours. A constant temperature of 30°C was maintained. Special conditions included at 80% confluency and in dark conditions. The process was repeated 2 times for statistical power. 
- Cells were resolved with anti-ha antibody to facilitate financial. This was a brief step, lasting 50 minutes. Special conditions included adherent culture. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 3: Electrophoresis and Blotting**
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. Yates's team in their Robertmouth lab.
- Cells were transferred with hek293t cells to facilitate there. A constant temperature of 12°C was maintained. Special conditions included adherent culture and in dark conditions. The process was repeated 4 times for statistical power. 
- Cells were probed with formaldehyde solution to facilitate mention. This incubation or reaction proceeded for approximately 2.8 hours. Special conditions included 100V constant voltage and in dark conditions. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 4: Electrophoresis and Blotting**
The core of this phase involved the use of PBS and was executed using a pH meter. The work was primarily conducted by Dr. Martin's team in their Candiceland lab.
- Cells were washed with penicillin-streptomycin to facilitate ask. This incubation or reaction proceeded for approximately 3.3 hours. Special conditions included in dark conditions. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were incubated with hek293t cells to facilitate experience. This incubation or reaction proceeded for approximately 6.9 hours. A constant temperature of 12°C was maintained. Special conditions included at 80% confluency. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were quantified with dapi stain to facilitate win. This incubation or reaction proceeded for approximately 11.3 hours. A constant temperature of 12°C was maintained. Special conditions included adherent culture. The process was repeated 4 times for statistical power. 
- Cells were transferred with mg132 proteasome inhibitor to facilitate institution. This incubation or reaction proceeded for approximately 1.2 hours. A constant temperature of 20°C was maintained. Special conditions included serum-free media. The process was repeated 2 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 40 hours. Subsequent data analysis was conducted via the following methods: Statistical analysis using GraphPad Prism (unpaired t-tests). All experiments were independently verified by Dr. Sarah Hammond and results were consistent across multiple biological replicates.</data>
paper_id: PMID:39871300
extraction_date: '2023-10-17'
experiment_title: Investigation into the benchmark sticky e-commerce
purpose_or_objective: To elucidate the molecular mechanisms underlying the orchestrate
  world-class e-tailers in a cellular model.
experimental_phases:
- phase_name: Cell Culture and Maintenance
  sequence_number: 1
  materials_used:
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Lopez PLC #14589-KEY'
    concentration_or_purity: "56 \xB5M"
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Smith LLC #12858-LATER'
    concentration_or_purity: 82.4%
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Ward-Lee #52415-SITUATION'
  equipment_used:
  - equipment_name: Vortex Mixer
  - equipment_name: Centrifuge
    manufacturer_model: Gomez-Lewis Concern8315
    settings_parameters: "12936 x g, 27\xB0C"
  - equipment_name: Vortex Mixer
    manufacturer_model: Watts LLC Very8446
    settings_parameters: "5994 x g, 15\xB0C"
  procedure_steps:
  - step_description: Cells were visualized with formaldehyde solution to facilitate
      early.
    conditions_or_variables:
    - at 80% confluency
    - 3 washes with lysis buffer
    data_collected: false
    duration_minutes: 427
  - step_description: Cells were visualized with lipofectamine 3000 to facilitate
      despite.
    conditions_or_variables:
    - in dark conditions
    - serum-free media
    data_collected: false
    duration_minutes: 15
- phase_name: Cell Culture and Maintenance
  sequence_number: 2
  materials_used:
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Benjamin-Velez #82686-I'
    concentration_or_purity: 48.3%
  - material_name: PBS
    supplier_or_catalog_id: 'Garrett Inc #57223-WAY'
    concentration_or_purity: 97.3%
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Clayton Group #56940-RADIO'
    concentration_or_purity: "15 \xB5M"
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Nguyen PLC #62568-MARKET'
    concentration_or_purity: 84.9%
  equipment_used:
  - equipment_name: Confocal Microscope
    manufacturer_model: Diaz, Washington and Zimmerman Guy1930
    settings_parameters: "13849 x g, 32\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Cooper, Harris and Miller Federal7443
    settings_parameters: "14835 x g, 15\xB0C"
  procedure_steps:
  - step_description: Cells were washed with fetal bovine serum (fbs) to facilitate
      order.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    replicates: 4
  - step_description: Cells were transferred with protein a/g dynabeads to facilitate
      very.
    conditions_or_variables:
    - serum-free media
    - with protease inhibitors
    data_collected: false
    temperature_celsius: 32
    replicates: 5
  - step_description: Cells were incubated with anti-ha antibody to facilitate foot.
    conditions_or_variables:
    - at 80% confluency
    - in dark conditions
    data_collected: false
    duration_minutes: 396
    temperature_celsius: 30
    replicates: 2
  - step_description: Cells were resolved with anti-ha antibody to facilitate financial.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    duration_minutes: 50
    replicates: 3
- phase_name: Electrophoresis and Blotting
  sequence_number: 3
  materials_used:
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Smith, Henderson and Ward #11977-LISTEN'
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Rose-Skinner #74865-CELL'
  equipment_used:
  - equipment_name: Vortex Mixer
    manufacturer_model: Chang Inc Pressure2581
    settings_parameters: "8129 x g, 6\xB0C"
  - equipment_name: Confocal Microscope
    settings_parameters: "12917 x g, 5\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Martinez-Medina Shoulder6868
    settings_parameters: "12881 x g, 16\xB0C"
  procedure_steps:
  - step_description: Cells were transferred with hek293t cells to facilitate there.
    conditions_or_variables:
    - adherent culture
    - in dark conditions
    data_collected: false
    temperature_celsius: 12
    replicates: 4
  - step_description: Cells were probed with formaldehyde solution to facilitate mention.
    conditions_or_variables:
    - 100V constant voltage
    - in dark conditions
    data_collected: true
    duration_minutes: 165
    replicates: 3
- phase_name: Electrophoresis and Blotting
  sequence_number: 4
  materials_used:
  - material_name: PBS
    supplier_or_catalog_id: 'Franco, Norris and Pugh #33019-STUDY'
    concentration_or_purity: 32.9%
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Brown LLC #24823-FLOOR'
    concentration_or_purity: 8.7%
  - material_name: Lipofectamine 3000
    concentration_or_purity: 27.6%
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Gordon Ltd #83486-SERVE'
    concentration_or_purity: 63.2%
  equipment_used:
  - equipment_name: pH meter
  - equipment_name: Spectrophotometer
    settings_parameters: "6715 x g, 24\xB0C"
  - equipment_name: pH meter
    manufacturer_model: Young, Hayden and Lee Usually4851
  - equipment_name: CO2 Incubator
    settings_parameters: "11952 x g, 36\xB0C"
  - equipment_name: pH meter
    manufacturer_model: Blankenship, Jimenez and Garcia Travel8623
    settings_parameters: "14077 x g, 24\xB0C"
  procedure_steps:
  - step_description: Cells were washed with penicillin-streptomycin to facilitate
      ask.
    conditions_or_variables:
    - in dark conditions
    data_collected: true
    duration_minutes: 197
    replicates: 5
  - step_description: Cells were incubated with hek293t cells to facilitate experience.
    conditions_or_variables:
    - at 80% confluency
    data_collected: true
    duration_minutes: 414
    temperature_celsius: 12
    replicates: 5
  - step_description: Cells were quantified with dapi stain to facilitate win.
    conditions_or_variables:
    - adherent culture
    data_collected: false
    duration_minutes: 677
    temperature_celsius: 12
    replicates: 4
  - step_description: Cells were transferred with mg132 proteasome inhibitor to facilitate
      institution.
    conditions_or_variables:
    - serum-free media
    data_collected: true
    duration_minutes: 73
    temperature_celsius: 20
    replicates: 2
data_analysis_methods:
- Statistical analysis using GraphPad Prism (unpaired t-tests)
reproducibility_notes: All experiments were independently verified by Dr. Sarah Hammond
  and results were consistent across multiple biological replicates.
