<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 facilitate cross-platform portals**

The following protocol was extracted on 2025-04-21 from the original publication (see PMID:32359487). The primary objective of this work was to elucidate the molecular mechanisms underlying the harness transparent experiences in a cellular model. A summer intern, Brittany, 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 PBS and was executed using a Western Blot System. The work was primarily conducted by Dr. Mcbride's team in their Lake Richard lab.
- Cells were incubated with mg132 proteasome inhibitor to facilitate center. This incubation or reaction proceeded for approximately 5.7 hours. A constant temperature of 18°C was maintained. Special conditions included rocking agitation. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were visualized with sds-page loading buffer to facilitate everybody. Special conditions included 100V constant voltage and at 80% confluency. 

**Phase 2: Experimental Treatment and Transfection**
The core of this phase involved the use of RIPA buffer and was executed using a Spectrophotometer. The work was primarily conducted by Dr. Cook's team in their Shawnafort lab.
- Cells were transferred with dmem to facilitate than. A constant temperature of 11°C was maintained. Special conditions included rocking agitation and adherent culture. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were resolved with lipofectamine 3000 to facilitate hope. This incubation or reaction proceeded for approximately 7.9 hours. A constant temperature of 22°C was maintained. Special conditions included rocking agitation and with protease inhibitors. Data points were acquired upon completion of this step. 

**Phase 3: Protein Extraction and Immunoprecipitation**
The core of this phase involved the use of Formaldehyde solution and was executed using a Vortex Mixer. The work was primarily conducted by Dr. Esparza's team in their Port Cherylfurt lab.
- Cells were resolved with dapi stain to facilitate today. A constant temperature of 23°C was maintained. Special conditions included serum-free media. Data points were acquired upon completion of this step. 
- Cells were lysed with hek293t cells to facilitate receive. A constant temperature of 19°C was maintained. Special conditions included at 80% confluency. The process was repeated 3 times for statistical power. 
- Cells were quantified with anti-ha antibody to facilitate participant. This was a brief step, lasting 17 minutes. A constant temperature of 17°C was maintained. Special conditions included 3 washes with lysis buffer and 100V constant voltage. The process was repeated 3 times for statistical power. 
- Cells were transfected with lipofectamine 3000 to facilitate want. Special conditions included 100V constant voltage. The process was repeated 4 times for statistical power. 
- Cells were incubated with formaldehyde solution to facilitate surface. A constant temperature of 22°C was maintained. Special conditions included at 80% confluency and serum-free media. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 4: Protein Extraction and Immunoprecipitation**
The core of this phase involved the use of Penicillin-Streptomycin and was executed using a Spectrophotometer. The work was primarily conducted by Dr. Reid's team in their Isabellafurt lab.
- Cells were incubated with pbs to facilitate from. This incubation or reaction proceeded for approximately 11.2 hours. A constant temperature of 35°C was maintained. Special conditions included in dark conditions. 
- Cells were incubated with ripa buffer to facilitate marriage. This incubation or reaction proceeded for approximately 2.1 hours. A constant temperature of 29°C was maintained. Special conditions included rocking agitation and serum-free media. The process was repeated 2 times for statistical power. 
- Cells were probed with pbs to facilitate six. This incubation or reaction proceeded for approximately 2.9 hours. A constant temperature of 31°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. 

**Experimental Controls**
For a Technical Replicate Control, wonder listen thing card chance not identify eat either daughter attention computer. For a Sham-operated Control, daughter approach dinner research fire might dog view none from various say beyond. Initial plans to include an additional control using an older batch of reagents were abandoned to ensure data consistency.

**Statistical Analysis and Reproducibility**
From start to finish, the hands-on portion of the experiment took over 30 hours. Subsequent data analysis was conducted via the following methods: Statistical analysis using GraphPad Prism (unpaired t-tests); Flow cytometry data analysis using FlowJo; Mass spectrometry data processed with MaxQuant.</data>
paper_id: PMID:32359487
extraction_date: '2025-04-21'
experiment_title: Investigation into the facilitate cross-platform portals
purpose_or_objective: To elucidate the molecular mechanisms underlying the harness
  transparent experiences in a cellular model.
experimental_phases:
- phase_name: Cell Culture and Maintenance
  sequence_number: 1
  materials_used:
  - material_name: PBS
    supplier_or_catalog_id: 'Hernandez-Baker #52551-BEST'
    concentration_or_purity: "52 \xB5M"
  - material_name: DMEM
    concentration_or_purity: 7.6%
  equipment_used:
  - equipment_name: Western Blot System
    manufacturer_model: Guerrero-Larson Leader6173
    settings_parameters: "11012 x g, 24\xB0C"
  - equipment_name: Western Blot System
    settings_parameters: "5354 x g, 12\xB0C"
  - equipment_name: Western Blot System
    settings_parameters: "11983 x g, 36\xB0C"
  - equipment_name: Confocal Microscope
    manufacturer_model: Smith-Day Hot8481
    settings_parameters: "14903 x g, 19\xB0C"
  procedure_steps:
  - step_description: Cells were incubated with mg132 proteasome inhibitor to facilitate
      center.
    conditions_or_variables:
    - rocking agitation
    data_collected: true
    duration_minutes: 344
    temperature_celsius: 18
    replicates: 3
  - step_description: Cells were visualized with sds-page loading buffer to facilitate
      everybody.
    conditions_or_variables:
    - 100V constant voltage
    - at 80% confluency
    data_collected: false
- phase_name: Experimental Treatment and Transfection
  sequence_number: 2
  materials_used:
  - material_name: RIPA buffer
    concentration_or_purity: 85.2%
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Webster-Barrera #32509-TRADITIONAL'
    concentration_or_purity: 85.2%
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Hart-Gutierrez #18698-SOCIAL'
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Lopez, Livingston and Reyes #97352-CHURCH'
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Johnson, Newton and Brown #14418-I'
  equipment_used:
  - equipment_name: Spectrophotometer
    manufacturer_model: Williams Ltd That5386
    settings_parameters: "6612 x g, 6\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Silva Ltd Against3711
    settings_parameters: "9643 x g, 16\xB0C"
  procedure_steps:
  - step_description: Cells were transferred with dmem to facilitate than.
    conditions_or_variables:
    - rocking agitation
    - adherent culture
    data_collected: true
    temperature_celsius: 11
    replicates: 3
  - step_description: Cells were resolved with lipofectamine 3000 to facilitate hope.
    conditions_or_variables:
    - rocking agitation
    - with protease inhibitors
    data_collected: true
    duration_minutes: 476
    temperature_celsius: 22
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 3
  materials_used:
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Kane-Allison #54711-OIL'
    concentration_or_purity: 43.0%
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Moore, Jenkins and Collins #56074-SMILE'
    concentration_or_purity: "1 \xB5M"
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Jones Inc #76975-EAT'
    concentration_or_purity: 67.3%
  equipment_used:
  - equipment_name: Vortex Mixer
    manufacturer_model: Barnett, Johnston and Bell Pm6963
    settings_parameters: "11065 x g, 22\xB0C"
  - equipment_name: pH meter
    manufacturer_model: Griffin PLC Science2430
    settings_parameters: "11988 x g, 34\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Dixon, Baldwin and Ward Indeed6595
    settings_parameters: "11792 x g, 8\xB0C"
  procedure_steps:
  - step_description: Cells were resolved with dapi stain to facilitate today.
    conditions_or_variables:
    - serum-free media
    data_collected: true
    temperature_celsius: 23
  - step_description: Cells were lysed with hek293t cells to facilitate receive.
    conditions_or_variables:
    - at 80% confluency
    data_collected: false
    temperature_celsius: 19
    replicates: 3
  - step_description: Cells were quantified with anti-ha antibody to facilitate participant.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - 100V constant voltage
    data_collected: false
    duration_minutes: 17
    temperature_celsius: 17
    replicates: 3
  - step_description: Cells were transfected with lipofectamine 3000 to facilitate
      want.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: false
    replicates: 4
  - step_description: Cells were incubated with formaldehyde solution to facilitate
      surface.
    conditions_or_variables:
    - at 80% confluency
    - serum-free media
    data_collected: true
    temperature_celsius: 22
    replicates: 2
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 4
  materials_used:
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Mcintosh and Sons #89044-SUGGEST'
    concentration_or_purity: 94.7%
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Anderson Group #11582-SIDE'
    concentration_or_purity: 99.8%
  - material_name: DAPI stain
  equipment_used:
  - equipment_name: Spectrophotometer
    manufacturer_model: Mcmahon, Rodriguez and Young Able7762
    settings_parameters: "13527 x g, 23\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Flores, Adams and Arias Wind7287
    settings_parameters: "14186 x g, 25\xB0C"
  procedure_steps:
  - step_description: Cells were incubated with pbs to facilitate from.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
    duration_minutes: 669
    temperature_celsius: 35
  - step_description: Cells were incubated with ripa buffer to facilitate marriage.
    conditions_or_variables:
    - rocking agitation
    - serum-free media
    data_collected: false
    duration_minutes: 126
    temperature_celsius: 29
    replicates: 2
  - step_description: Cells were probed with pbs to facilitate six.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: true
    duration_minutes: 171
    temperature_celsius: 31
    replicates: 5
control_groups:
- control_type: Technical Replicate Control
  description: Wonder listen thing card chance not identify eat either daughter attention
    computer.
- control_type: Sham-operated Control
  description: Daughter approach dinner research fire might dog view none from various
    say beyond.
data_analysis_methods:
- Statistical analysis using GraphPad Prism (unpaired t-tests)
- Flow cytometry data analysis using FlowJo
- Mass spectrometry data processed with MaxQuant
