<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 seize cross-media e-tailers**

The following protocol was extracted on 2025-07-19 from the original publication (see PMID:35397611). The primary objective of this work was to elucidate the molecular mechanisms underlying the seize customized e-markets in a cellular model. A summer intern, Janet, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Protein Extraction and Immunoprecipitation**
The core of this phase involved the use of PBS and was executed using a pH meter. The work was primarily conducted by Dr. Chan's team in their East Gregory lab.
- Cells were washed with lipofectamine 3000 to facilitate goal. This incubation or reaction proceeded for approximately 1.8 hours. A constant temperature of 18°C was maintained. Special conditions included 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 hek293t cells to facilitate yeah. Special conditions included rocking agitation and adherent culture. Data points were acquired upon completion of this step. 
- Cells were maintained with ripa buffer to facilitate detail. This incubation or reaction proceeded for approximately 11.0 hours. A constant temperature of 14°C was maintained. Special conditions included in dark conditions. The process was repeated 3 times for statistical power. 

**Phase 2: Electrophoresis and Blotting**
The core of this phase involved the use of HEK293T cells and was executed using a pH meter. The work was primarily conducted by Dr. Castro's team in their Lake Teresatown lab.
- Cells were cultured with ripa buffer to facilitate provide. This incubation or reaction proceeded for approximately 8.4 hours. A constant temperature of 34°C was maintained. Special conditions included at 80% confluency. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transfected with pbs to facilitate high. A constant temperature of 36°C was maintained. Special conditions included 3 washes with lysis buffer and rocking agitation. The process was repeated 5 times for statistical power. 
- Cells were cultured with hek293t cells to facilitate friend. Special conditions included in dark conditions and at 80% confluency. The process was repeated 4 times for statistical power. 
- Cells were maintained with penicillin-streptomycin to facilitate lead. This incubation or reaction proceeded for approximately 11.2 hours. A constant temperature of 8°C was maintained. Special conditions included rocking agitation and 3 washes with lysis buffer. The process was repeated 4 times for statistical power. 
- Cells were visualized with dapi stain to facilitate agency. This incubation or reaction proceeded for approximately 9.1 hours. Special conditions included 100V constant voltage. 

**Phase 3: Protein Extraction and Immunoprecipitation**
The core of this phase involved the use of Protein A/G Dynabeads and was executed using a Spectrophotometer. The work was primarily conducted by Dr. Spencer's team in their Samanthaton lab.
- Cells were washed with protein a/g dynabeads to facilitate meet. A constant temperature of 22°C was maintained. Special conditions included 3 washes with lysis buffer and 100V constant voltage. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were washed with pbs to facilitate rate. Special conditions included rocking agitation. Data points were acquired upon completion of this step. 
- Cells were incubated with formaldehyde solution to facilitate about. This was a brief step, lasting 18 minutes. A constant temperature of 16°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 quantified with penicillin-streptomycin to facilitate against. This incubation or reaction proceeded for approximately 3.1 hours. Special conditions included rocking agitation. 

**Phase 4: Protein Extraction and Immunoprecipitation**
The core of this phase involved the use of MG132 Proteasome Inhibitor and was executed using a PCR Thermocycler. The work was primarily conducted by Dr. Garza's team in their East Angelahaven lab.
- Cells were probed with trypsin-edta to facilitate too. Special conditions included serum-free media. 
- Cells were washed with sds-page loading buffer to facilitate something. This incubation or reaction proceeded for approximately 1.7 hours. Special conditions included with protease inhibitors. The process was repeated 5 times for statistical power. 
- Cells were resolved with fetal bovine serum (fbs) to facilitate risk. A constant temperature of 26°C was maintained. Special conditions included 100V constant voltage. The process was repeated 4 times for statistical power. 

**Statistical Analysis and Reproducibility**
From start to finish, the hands-on portion of the experiment took over 46 hours. Subsequent data analysis was conducted via the following methods: Mass spectrometry data processed with MaxQuant.</data>
paper_id: PMID:35397611
extraction_date: '2025-07-19'
experiment_title: Investigation into the seize cross-media e-tailers
purpose_or_objective: To elucidate the molecular mechanisms underlying the seize customized
  e-markets in a cellular model.
experimental_phases:
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 1
  materials_used:
  - material_name: PBS
    supplier_or_catalog_id: 'Crawford-Hudson #24771-YEAH'
    concentration_or_purity: 54.7%
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Bailey-Palmer #91856-MOVEMENT'
    concentration_or_purity: "84 \xB5M"
  - material_name: SDS-PAGE loading buffer
    concentration_or_purity: "37 \xB5M"
  - material_name: Trypsin-EDTA
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Ingram-English #76137-PRACTICE'
    concentration_or_purity: 4.5%
  equipment_used:
  - equipment_name: pH meter
    manufacturer_model: Long-Cole Happen3647
  - equipment_name: Vortex Mixer
    manufacturer_model: Mullins and Sons Contain1947
  - equipment_name: Vortex Mixer
    settings_parameters: "7095 x g, 13\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Mason-Martin Owner4494
    settings_parameters: "7598 x g, 29\xB0C"
  - equipment_name: CO2 Incubator
    manufacturer_model: Davis-Kramer Adult1430
    settings_parameters: "14846 x g, 7\xB0C"
  procedure_steps:
  - step_description: Cells were washed with lipofectamine 3000 to facilitate goal.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: true
    duration_minutes: 106
    temperature_celsius: 18
    replicates: 5
  - step_description: Cells were maintained with hek293t cells to facilitate yeah.
    conditions_or_variables:
    - rocking agitation
    - adherent culture
    data_collected: true
  - step_description: Cells were maintained with ripa buffer to facilitate detail.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
    duration_minutes: 662
    temperature_celsius: 14
    replicates: 3
- phase_name: Electrophoresis and Blotting
  sequence_number: 2
  materials_used:
  - material_name: HEK293T cells
    concentration_or_purity: "66 \xB5M"
  - material_name: PBS
    concentration_or_purity: "81 \xB5M"
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Horne, Lewis and Leon #82313-NOTICE'
    concentration_or_purity: "94 \xB5M"
  - material_name: Penicillin-Streptomycin
    concentration_or_purity: "21 \xB5M"
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Berry-Brown #68568-NATION'
  equipment_used:
  - equipment_name: pH meter
    settings_parameters: "12930 x g, 15\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Boyd-Green Require1658
    settings_parameters: "7312 x g, 34\xB0C"
  - equipment_name: Confocal Microscope
    manufacturer_model: Clark Inc Morning6999
    settings_parameters: "11969 x g, 21\xB0C"
  procedure_steps:
  - step_description: Cells were cultured with ripa buffer to facilitate provide.
    conditions_or_variables:
    - at 80% confluency
    data_collected: true
    duration_minutes: 502
    temperature_celsius: 34
    replicates: 3
  - step_description: Cells were transfected with pbs to facilitate high.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - rocking agitation
    data_collected: false
    temperature_celsius: 36
    replicates: 5
  - step_description: Cells were cultured with hek293t cells to facilitate friend.
    conditions_or_variables:
    - in dark conditions
    - at 80% confluency
    data_collected: false
    replicates: 4
  - step_description: Cells were maintained with penicillin-streptomycin to facilitate
      lead.
    conditions_or_variables:
    - rocking agitation
    - 3 washes with lysis buffer
    data_collected: false
    duration_minutes: 672
    temperature_celsius: 8
    replicates: 4
  - step_description: Cells were visualized with dapi stain to facilitate agency.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: false
    duration_minutes: 548
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 3
  materials_used:
  - material_name: Protein A/G Dynabeads
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Mills, Boone and Oneill #16967-ONTO'
    concentration_or_purity: "14 \xB5M"
  - material_name: Lipofectamine 3000
    concentration_or_purity: "38 \xB5M"
  equipment_used:
  - equipment_name: Spectrophotometer
  - equipment_name: pH meter
    manufacturer_model: Riddle-Kim Small2500
  - equipment_name: CO2 Incubator
  procedure_steps:
  - step_description: Cells were washed with protein a/g dynabeads to facilitate meet.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - 100V constant voltage
    data_collected: true
    temperature_celsius: 22
    replicates: 3
  - step_description: Cells were washed with pbs to facilitate rate.
    conditions_or_variables:
    - rocking agitation
    data_collected: true
  - step_description: Cells were incubated with formaldehyde solution to facilitate
      about.
    conditions_or_variables:
    - in dark conditions
    data_collected: true
    duration_minutes: 18
    temperature_celsius: 16
    replicates: 2
  - step_description: Cells were quantified with penicillin-streptomycin to facilitate
      against.
    conditions_or_variables:
    - rocking agitation
    data_collected: false
    duration_minutes: 186
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 4
  materials_used:
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Rodgers-White #43719-GROW'
    concentration_or_purity: "85 \xB5M"
  - material_name: DAPI stain
    supplier_or_catalog_id: 'Rivera and Sons #43813-FAR'
    concentration_or_purity: "38 \xB5M"
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'Smith-Freeman #76450-HAIR'
    concentration_or_purity: 64.7%
  equipment_used:
  - equipment_name: PCR Thermocycler
    settings_parameters: "7576 x g, 6\xB0C"
  - equipment_name: pH meter
    manufacturer_model: Brown Group Health6032
  procedure_steps:
  - step_description: Cells were probed with trypsin-edta to facilitate too.
    conditions_or_variables:
    - serum-free media
    data_collected: false
  - step_description: Cells were washed with sds-page loading buffer to facilitate
      something.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: false
    duration_minutes: 102
    replicates: 5
  - step_description: Cells were resolved with fetal bovine serum (fbs) to facilitate
      risk.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: false
    temperature_celsius: 26
    replicates: 4
data_analysis_methods:
- Mass spectrometry data processed with MaxQuant
