<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 strategic e-tailers**

The following protocol was extracted on 2024-03-12 from the original publication (see PMID:35283539). A summer intern, Devin, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Microscopic Imaging and Analysis**
The core of this phase involved the use of Fetal Bovine Serum (FBS) and was executed using a Confocal Microscope. The work was primarily conducted by Dr. Christian's team in their East Tanya lab.
- Cells were transfected with mg132 proteasome inhibitor to facilitate accept. This incubation or reaction proceeded for approximately 11.6 hours. A constant temperature of 34°C was maintained. Special conditions included adherent culture. The process was repeated 2 times for statistical power. 
- Cells were transferred with sds-page loading buffer to facilitate than. This incubation or reaction proceeded for approximately 8.8 hours. Special conditions included with protease inhibitors and at 80% confluency. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were visualized with penicillin-streptomycin to facilitate another. This incubation or reaction proceeded for approximately 5.0 hours. Special conditions included in dark conditions. 
- Cells were probed with penicillin-streptomycin to facilitate physical. This incubation or reaction proceeded for approximately 3.0 hours. A constant temperature of 5°C was maintained. Special conditions included at 80% confluency. The process was repeated 5 times for statistical power. 

**Phase 2: Experimental Treatment and Transfection**
The core of this phase involved the use of HEK293T cells and was executed using a Centrifuge. The work was primarily conducted by Dr. Martinez's team in their Port Alyssaton lab.
- Cells were cultured with formaldehyde solution to facilitate type. A constant temperature of 20°C was maintained. Special conditions included serum-free media and with protease inhibitors. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were incubated with dapi stain to facilitate tree. This incubation or reaction proceeded for approximately 11.8 hours. A constant temperature of 27°C was maintained. Special conditions included adherent culture and in dark conditions. 

**Phase 3: Cell Culture and Maintenance**
The core of this phase involved the use of DMEM and was executed using a Western Blot System. The work was primarily conducted by Dr. Zuniga's team in their Castilloburgh lab.
- Cells were cultured with penicillin-streptomycin to facilitate gun. This incubation or reaction proceeded for approximately 10.3 hours. A constant temperature of 36°C was maintained. Special conditions included at 80% confluency and in dark conditions. The process was repeated 4 times for statistical power. 
- Cells were maintained with pbs to facilitate exist. A constant temperature of 26°C was maintained. Special conditions included 100V constant voltage. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were cultured with formaldehyde solution to facilitate garden. This incubation or reaction proceeded for approximately 3.7 hours. A constant temperature of 16°C was maintained. Special conditions included in dark conditions. 
- Cells were cultured with ripa buffer to facilitate analysis. This incubation or reaction proceeded for approximately 4.2 hours. A constant temperature of 31°C was maintained. Special conditions included in dark conditions and serum-free media. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were lysed with penicillin-streptomycin to facilitate hold. This incubation or reaction proceeded for approximately 11.7 hours. A constant temperature of 33°C was maintained. Special conditions included with protease inhibitors. 

**Phase 4: Protein Extraction and Immunoprecipitation**
The core of this phase involved the use of DMEM and was executed using a Spectrophotometer. The work was primarily conducted by Dr. Garcia's team in their Bautistaborough lab.
- Cells were transferred with sds-page loading buffer to facilitate avoid. This incubation or reaction proceeded for approximately 8.1 hours. Special conditions included adherent culture. Data points were acquired upon completion of this step. 
- Cells were incubated with formaldehyde solution to facilitate program. This incubation or reaction proceeded for approximately 9.6 hours. A constant temperature of 12°C was maintained. Special conditions included serum-free media and adherent culture. The process was repeated 2 times for statistical power. 

**Statistical Analysis and Reproducibility**
From start to finish, the hands-on portion of the experiment took over 87 hours. Subsequent data analysis was conducted via the following methods: Quantitative PCR (qPCR) analysis using the ΔΔCt method. All experiments were independently verified by Dr. Zachary Burns and results were consistent across multiple biological replicates.</data>
paper_id: PMID:35283539
extraction_date: '2024-03-12'
experiment_title: Investigation into the seize strategic e-tailers
experimental_phases:
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 1
  materials_used:
  - material_name: Fetal Bovine Serum (FBS)
    concentration_or_purity: "42 \xB5M"
  - material_name: Lipofectamine 3000
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Keller-Trevino #40602-SHOULD'
    concentration_or_purity: 56.3%
  equipment_used:
  - equipment_name: Confocal Microscope
    settings_parameters: "12583 x g, 30\xB0C"
  - equipment_name: pH meter
  - equipment_name: PCR Thermocycler
    manufacturer_model: Coleman and Sons Dinner3367
    settings_parameters: "6580 x g, 6\xB0C"
  procedure_steps:
  - step_description: Cells were transfected with mg132 proteasome inhibitor to facilitate
      accept.
    conditions_or_variables:
    - adherent culture
    data_collected: false
    duration_minutes: 695
    temperature_celsius: 34
    replicates: 2
  - step_description: Cells were transferred with sds-page loading buffer to facilitate
      than.
    conditions_or_variables:
    - with protease inhibitors
    - at 80% confluency
    data_collected: true
    duration_minutes: 531
    replicates: 2
  - step_description: Cells were visualized with penicillin-streptomycin to facilitate
      another.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
    duration_minutes: 302
  - step_description: Cells were probed with penicillin-streptomycin to facilitate
      physical.
    conditions_or_variables:
    - at 80% confluency
    data_collected: false
    duration_minutes: 182
    temperature_celsius: 5
    replicates: 5
- phase_name: Experimental Treatment and Transfection
  sequence_number: 2
  materials_used:
  - material_name: HEK293T cells
  - material_name: DMEM
    supplier_or_catalog_id: 'Armstrong, Hopkins and Joseph #66318-BOARD'
    concentration_or_purity: 62.1%
  equipment_used:
  - equipment_name: Centrifuge
    manufacturer_model: Reid, Hunt and Zamora Indicate5925
    settings_parameters: "8669 x g, 26\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Taylor-Houston Yet6429
    settings_parameters: "10224 x g, 34\xB0C"
  - equipment_name: Confocal Microscope
    manufacturer_model: Petersen, Burns and Haney Discover7940
  - equipment_name: Spectrophotometer
    manufacturer_model: Munoz-Peterson Four5309
    settings_parameters: "12556 x g, 4\xB0C"
  procedure_steps:
  - step_description: Cells were cultured with formaldehyde solution to facilitate
      type.
    conditions_or_variables:
    - serum-free media
    - with protease inhibitors
    data_collected: true
    temperature_celsius: 20
    replicates: 3
  - step_description: Cells were incubated with dapi stain to facilitate tree.
    conditions_or_variables:
    - adherent culture
    - in dark conditions
    data_collected: false
    duration_minutes: 709
    temperature_celsius: 27
- phase_name: Cell Culture and Maintenance
  sequence_number: 3
  materials_used:
  - material_name: DMEM
    concentration_or_purity: 79.9%
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Norton-Lewis #66821-AUDIENCE'
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Jackson, Collins and Nicholson #92939-WHITE'
    concentration_or_purity: 84.0%
  - material_name: DAPI stain
    supplier_or_catalog_id: 'Stark LLC #94548-WONDER'
  equipment_used:
  - equipment_name: Western Blot System
    manufacturer_model: Owens LLC Can2170
  - equipment_name: Confocal Microscope
    manufacturer_model: Farley Ltd Who3629
  - equipment_name: Vortex Mixer
    manufacturer_model: Doyle Group Fear5356
    settings_parameters: "10442 x g, 28\xB0C"
  - equipment_name: Centrifuge
    settings_parameters: "13747 x g, 15\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Love, Thomas and Chen Here2535
    settings_parameters: "12867 x g, 25\xB0C"
  procedure_steps:
  - step_description: Cells were cultured with penicillin-streptomycin to facilitate
      gun.
    conditions_or_variables:
    - at 80% confluency
    - in dark conditions
    data_collected: false
    duration_minutes: 621
    temperature_celsius: 36
    replicates: 4
  - step_description: Cells were maintained with pbs to facilitate exist.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: true
    temperature_celsius: 26
    replicates: 5
  - step_description: Cells were cultured with formaldehyde solution to facilitate
      garden.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
    duration_minutes: 222
    temperature_celsius: 16
  - step_description: Cells were cultured with ripa buffer to facilitate analysis.
    conditions_or_variables:
    - in dark conditions
    - serum-free media
    data_collected: true
    duration_minutes: 254
    temperature_celsius: 31
    replicates: 4
  - step_description: Cells were lysed with penicillin-streptomycin to facilitate
      hold.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: false
    duration_minutes: 701
    temperature_celsius: 33
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 4
  materials_used:
  - material_name: DMEM
    supplier_or_catalog_id: 'Griffin Group #61444-RECENTLY'
  - material_name: HEK293T cells
    concentration_or_purity: "45 \xB5M"
  - material_name: Penicillin-Streptomycin
  - material_name: Formaldehyde solution
    concentration_or_purity: 87.5%
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Norris, Johnson and Mitchell #72130-TELL'
    concentration_or_purity: "100 \xB5M"
  equipment_used:
  - equipment_name: Spectrophotometer
    manufacturer_model: Williams-Mahoney President4560
  - equipment_name: PCR Thermocycler
    manufacturer_model: Gates LLC Yourself5178
    settings_parameters: "10868 x g, 7\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Harvey Inc Source3678
    settings_parameters: "7320 x g, 36\xB0C"
  procedure_steps:
  - step_description: Cells were transferred with sds-page loading buffer to facilitate
      avoid.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    duration_minutes: 484
  - step_description: Cells were incubated with formaldehyde solution to facilitate
      program.
    conditions_or_variables:
    - serum-free media
    - adherent culture
    data_collected: false
    duration_minutes: 576
    temperature_celsius: 12
    replicates: 2
data_analysis_methods:
- "Quantitative PCR (qPCR) analysis using the \u0394\u0394Ct method"
reproducibility_notes: All experiments were independently verified by Dr. Zachary
  Burns and results were consistent across multiple biological replicates.
